A condensatopathy is an aberration of a condensate that drives a specific disease phenotype (refer to Glossary for condensate science terminology). The condensatopathy encyclopedia is a literature-based, semi-manually curated list of condensate phenotypes associated with disease pathophysiology supported by experimental evidence in relevant cellular, animal and/or clinical models.
NEW! The condensatopathy encyclopedia is now integrated in CD-CODE and cross-referenced with other large databases, including UniProt and Disease Ontology.
Curator in residence: Allysa P. Kemraj
The legacy condensatopathies table below will be retired by the end of the year.
Last updated on October 8, 2025 | N = 261
| Condensate name | Disease association | Dysregulation type | Reference |
|---|---|---|---|
| LANA body | Karposi Sarcoma herpresvirus infection | LANA bodies are viral episomes, located in the nucleus of the host cell, containing viral genome and the KSHV protein LANA, which form via phase separation. LANA bodies are dynamic structures that change during the cell cycle and viral life cycle. The host protein DAXX localizes to LANA bodies under latent infection, and it is ejected during the transition to lytic replication. LANA bodies form ring-like microscopic structure that associate with KSHV DNA during lytic reactivation. 1,6-hexanediol treatment partially dissolves LANA bodies and alters KSHV genome conformation. | https://pubmed.ncbi.nlm.nih.gov/33471863/ |
| Negri Body | Rabies virus (RABV) infection | Negri bodies are viral factories containing viral proteins P and N and form in the cytoplasm of cells infected with the Rabies virus (RABV). They exhibit liquid-like properties as evidenced by fusion observed with live imaging, and P protein fluoerescence recovers within seconds in FRAP experiments. Negri bodies are not miscible with stress granules. | https://pubmed.ncbi.nlm.nih.gov/28680096/ |
| Nucleolus | Cancer | Nucleoli are enlarged in rapidly proliferating cells, as measured by AgNOR staining, which stains for Ag-binding proteins localized to the nucleolous, primarily NPM1 and nucleolin. The number and histologic pattern of AgNOR staining allows patient stratification into high risk and low risk, across multiple types of cancer. | https://pubmed.ncbi.nlm.nih.gov/10588059/ |
| Nucleolus | Acute myeloid leukemia (AML) | NPM1c relocalizes from the nucleolus to the cytoplasm. Return of NPMc to the nucleus via gene editing induces cell growth arrest and differentiation in AML cultured cell lines OCI-AML3 and IMS-M2 and in primary AML cells. Targeted degradation of NPM1c, or treatment with the XPO1 inhibitor KPT-330/selinexor downreagulate HOX/Meis1 gene expression, induce cell growth and differentiation in AML cultured cells lines OCI-AML3 anf IMS-S2. Treatment of NPM1c/FLT3-IDT leukemic mice with KPT-330/selinexor prolonged animal survival, reduced leukemic liver infiltration, spleen wieghts and white blood cell counts. | https://pubmed.ncbi.nlm.nih.gov/17546053/ |
| Nucleolus | Neurodegeneration | polyGly-Arg (GR) and polyPro-Arg (PR) dipeptides, formed through translation of C9ORF72 repeat expansion, localize to nucleolar substructures and impair biophysical properties of NPM1 in vitro, and disrupt nucleolar dynamics and function in HeLa cells. Additionally, GR and PR induce spontaneous assembly of poorly dynamic stress granules and impair translation. GR and PR directly interact with LCD-containing RNA-binding proteins and alter their biophysical properties, disrupting ribosome biogenesis. | https://pubmed.ncbi.nlm.nih.gov/27768896/ |
| Nucleolus | Coronaviruses | Transfected coronavirus nucleoproteins from TGEV and MHV localize to nucleoli and disrupt cell division. | https://pubmed.ncbi.nlm.nih.gov/11533198/ |
| PML bodies | Human adenovirus infection | Adenovirus capsid protein VI accumulates into PML bodies upon viral entry in infected U2OS cells and displaces Daxx from PML bodies and translocates it into the cytoplasm, to I nhibit Daxx-mediated repression of the immediate early Ad E1A promoter.. | https://pubmed.ncbi.nlm.nih.gov/22427750/ |
| Stress granules | Dilated cardiomyopathy (DCM) | RNA-binding protein RBM20 undergoes mislocalization from the nucleus to the cytoplasm, where it forms aberrant ribonucleoprotein (RNP) condensates, in human iPSC-derived cardyomyocytes. These condensates resemble stress granules but are persistent and misregulated, disrupting RBM20’s normal role in nuclear RNA splicing. Cytoplasmic condensates formed by mutant RBM20 in tranfected U2OS cells recover within seconds in FRAP experiments and exhibit wetting and beading properties on the nucleus surface. | https://pubmed.ncbi.nlm.nih.gov/33188278/ |
| C9Orf72 RNA repeat foci | Amyotrophic lateral sclerosis (ALS) | Aberrant G4C2 repeat RNA, associated with ALS, form in vitro condensates in a concentration-dependent manner, with emergent properties dependent on the repeat length, ranging from spherical condensates to mech-like networks. Expression of G4C2 repeats in U2OS cells caused formation of nuclear puncta, that exhibit partial recovery after photobleaching (FRAP). | https://pubmed.ncbi.nlm.nih.gov/28562589/ |
| CAG RNA repeat foci | Huntington's disease (HD)|spinocerebellar ataxis (SCA) | Aberrant CAG repeat RNA, associated with Huntingtin's disease (HD) and spinocerebellar ataxia (SCA), form nuclear foci that co-localize with nuclear speckles in transfected, cultured U2OS cells and patient-derived fibroblasts. The nuclear condensates recover after photobleaching (FRAP) within minutes and demonstrate fusion in live imaging. The repeat RNA forms gel-like in vitro condensates, mediated by base-pairing. | https://pubmed.ncbi.nlm.nih.gov/28562589/ |
| CUG RNA repeat foci | Myotonic dystrophy (DM1) | Aberrant CUG repeat RNAs, associated with Myotonic dystrophy (DM1) form nuclear condensates in DM1 patient fibroblasts, and gel-like condensates in vitro. | https://pubmed.ncbi.nlm.nih.gov/28562589/ |
| EWS1-FLI condensate | Ewing's sarcoma | Aberrant transcriptional condensates formed by EWS-FLI1 fusion oncogene induces gene activation at GGAA microsatelite-target genes and drive oncogenic transformation in cultured cells. EWS-FLI1 condensates are dynamic, as demonstrated by fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS) and live-cell particle tracking (SPT). | https://pubmed.ncbi.nlm.nih.gov/29930090/ |
| NUP98-KDM5A condensates | Acute erythroid leukemia (AEL); acute myeloid leukemia (AML) | NUP98-KDM5A forms aberrant nuclear transcriptional condensates, as demonstrated by LC-MS and biCon-MS. NUP98-KDM5A condensates induce leukemogenic gene expression programs in hematopoietic progenitor cells. | https://pubmed.ncbi.nlm.nih.gov/33479542/ |
| Stress granules | Concurrent intrahepatic cholangiocarcinoma (ICC) and hepatolithiasis | E. Coli infection promotes cGLIS3 circularization and assembly of stress granules (SGs), which in turn promotes ICC progression, demonstrated in colony formation assays and in mouse models of lung metastasis. Hepatolithiasis-ICC patients express higher level of cGLIS3 and poorer prognosis. Mechanisticallty, cGLIS3 forms a complex with hnRNPA1 and G3BP1 to promote SG and activate NF-kB signaling pathway. | https://pubmed.ncbi.nlm.nih.gov/38368261/ |
| Nuclear speckles | Pancreatic ductal adenocarcinoma (PDAC)|Drug resistance | Analysis of a large-scale integrated dataset of human PDACs and a panel of 12 human PDAC cell lines, demonstrated that SIRT6 gene expression levels were higher in classical PDAC compared to basal PDAC samples. SIRT6 protein expression ATF4 is stabilized, and protected from degradation, and relocalized to nuclear speckles by binding to SIRT6 in Panc3.27 cell lines overexpressing SIRT6 and ATF4. In PDX models, tumor growth of basal but nor classical tumors was inhibited by treatment with flavopiridol, an ATP-competitive CDK9 inhibitor). | https://pubmed.ncbi.nlm.nih.gov/37134154/ |
| Stress granules | Age-related osteoporosis | Expression of HuR levels and HuR-positive stress granules (SG) positively associate with bone formation, with lower levels observed in older patients. HuR overexpression increased G3BP1 and Colla1 gene and protein expression and promoted SG formation in MC3T3-E1 cells. HuR overexpression increased G3BP1 and Runx2 gene and protein expression, promoted SG formation and induced osteogenic markers in mouse primary osteoblasts. HuR overexpression increased mRNA and protein levels of β-catenin, recruits β-catenin into SGs in MEC3T3-E1 cells. to facilitate osteogenesis in aging. Treatment with HuR activator, apigenin, enhances HuR expression, SG induction and recruitment of β-catenin and promotes osteoblast differentiation in MC3T3-E1 cells. In vivo, in OVX mouse models, treatment with apigenin promoted HuR, G3BP1 mRNA and protein expression, osteogenic gene expression and bone formation. | https://pubmed.ncbi.nlm.nih.gov/38375951/ |
| SHP2 condensates | SHP2‒associated cancers | SHP2-E76K mutation promotes formation of aberrant SHP2 condensates, hyperactivation of mitochondrial complex I/III and hyperactive mitichondrial metabolisn. Expression of SHP2 mutant causes malignant transformation into sarcoma-like stem cells of mouse mesenchymal stem cells (MSCs). Mice subcutaneously injected with MSCs expressing mutant SHP2, but not wild-type, form tumors in vivo. | https://pubmed.ncbi.nlm.nih.gov/38451719/ |
| ATG4B condensates | Gastric cancer|Metastasis | Circular RNA circSPECC1 directly interacts with ATG4B, promoting formation of cytoplasmic ATG4B condensates in gastric cancer (GC) cells, to drive ubiquitination and degradation of ATG4B and suppresses autophagy. This mechanism promotes metastasis in GC patient tissues. ATG4B recovers from photobleaching (FRAP) on the seconds timescale in cytoplasmic condensates formed in GC cells. | https://pubmed.ncbi.nlm.nih.gov/38433354/ |
| Stress granules | KrasG12D-dependent tumors|Pancreatic ductal adenocarcinoma (PDAC) | NUPR1 undergoes LLPS in vitro homotypically and heterotypically, in the presence of RNA. NUPR1 colocalizes with G3BP1 in arsenate-induced stress granules (SGs), and promotes induction of a larger number of SGs per cell, compared to cells with NUPR1 knock-down. KRAS-G12D mutation induces overexpression of NUPR1 and formation of NUPR1-dependent SGs in pancreatic cancer cells. Inihbition of NUPR1 with ZZW-115 induces cell death in KRAS-G12D-activated cells. ZZW-115 treatment prevents NUPR1-dependent SG induction and inhibits PDAC development in vivo in Pdx1-cre;LSL-Kras-G12D mouse models. | https://pubmed.ncbi.nlm.nih.gov/38360999/ |
| TDP-43 puncta | Amyotrophic lateral sclerosis (ALS) | TDP-43 co-localizes with in puncta with tunneling nanotubes (TNTs) -- actin cytoskeleton alterations, detected by F-actin staining, in lymphoblasts harvested from sporadic ALS patients. TDP-43 from diseased cells is secreted into the extracellular medium and spread the pathology to healthy myoblast cells treated with conditioned medium from diseased lymphoblasts, as measured by increase in cytoplasmic TDP-43 and TDP-43 phosphorylation and cytoskeleton changes. U2OS cells treated with conditioned medium from diseased cells showed cytoplasmic accumulation of TDP-43 and aggregation, and disruption of kinesin:tubulin interaction. | https://pubmed.ncbi.nlm.nih.gov/38325718/ |
| Stress granules | Drug resistance | Treatment of ovarian cancer cells A2780 with the PI3K inhibitor PKI-402 induces stress granule (SG) formation, activated mitochondrial unfolded protein response and mitochondrial function through relocalization of ATF5 into the nucleus. | https://pubmed.ncbi.nlm.nih.gov/38238825/ |
| Nucleolus | Cancer | NONO relocalizes to the nucleolus in response to DNA damage induced by etoposide treatment, in U2OS cells, reduces NONO occupancy at protein-coding gene promoters, to attentuate pre-mRNA synthesis and impair the formation of excessive R-loops and facilitate efficient NHEJ, thereby interfering with the double stranded break signaling. The mechanism is dependent on the RNA-binding activity of NONO RRM1 domain, and RNAPII-dependent expression of nucleolar transcripts, asincRNAs, at distinct IGS loci. NONO relocalization to the nucleolus mediates accumulation of pre-mRNA in the nucleolus, including CDKN1A transcript. | https://pubmed.ncbi.nlm.nih.gov/38224452/ |
| PML bodies | Pediatric gliomas | Histone 3.3 (H3.3) colocalizes with PML across the genome. In pediatric glioma, the presence of the K27M/G34R mutations in H3.3 disrupt PML bodies, and impairs glial differentiation in patinet-derived glioma cells. IDH1 R132H mutations in glial and myeloid malignancies also disrupt PML formation in oligodendroglioma cells. | https://pubmed.ncbi.nlm.nih.gov/38066546/ |
| EphA2 membrane condensates | Colorectal cancer | EphA2 expression is elevated in colorectal cancer (CRC) patient and the expression levels correlate with poor prognosis. Overexpression of EphA2 in HEK293 cells leads to formation of condensates on the plasma membrane. The condensates exhibit fusion and recovery after photobleaching (FRAP) on the tens of seconds timescale. RNA-seq profiling of CRC patient samples demonstrates that EphA2 expression levels positively correlate with expression of ferroptosis- and tumor-development-related genes, as well as macrophage, neutrophil and myeloid dendritic cell infiltration. | https://pubmed.ncbi.nlm.nih.gov/37980165/ |
| YTHDF1-ribosome condensates | Fragile X syndrome (FXS) | YTHDF1 forms cytoplasmic condensates in depolarized cultured mouse cortical neurons. Condensation of YTHDF1 is promoted by phosphorylated FMRP and co-condenses with with ribosomes to promote translation. Supressed formation of YTHD1 cytoplasmic condensates by salvianolic acid C (SAC) inhibits hyperactive YTHDF1 in neurons and reverses neurodevelopmental deficits in FXS mouse forebrain organoids. | https://pubmed.ncbi.nlm.nih.gov/37949069/ |
| PARIS aggregates | Parkinson's disease | Recombinant PARIS (ZNF746) phase separates in vitro, promoted by high ionic strength buffer conditions, and exhibit liquid-like behavior by undergoing fusion and fission, and recovers within seconds from photobleaching. At higher protein concentrations or in the rpesence of crowder, PARIS condensates transition to solid-like aggregates, with limited recovery from photobleaching and irregulat shapes. The low complexity fomain (LCD) is required for phase separation of PARIS, in vitro. PAR binding to the Zn-finger domain of PARIS promotes phase separation in vitro and enhances aggegation. In vivo, levels of PARIS increase with age, while levels of parkin decrease; PARIS accumulates and aggegates in the aging mice's brain in a PAR-dependent manner. PARIS aggregation by PAR‐activating α‐syn PFFs causes PGC‐1α reduction and mitochondrial dysfunction, contributing to DA neuronal death in a sporadic Parkinson's disease mouse model. | https://pubmed.ncbi.nlm.nih.gov/37870275/ |
| P bodies | Drug resistance | Dissolution of P bodies (decrease in size and number) in response to treatment with MEKi inhibitors leads to increase in KRAS and NRAS ongogene translation. P body protein levels inversely correlate with NRAS expression levels, and MAPK signaling promotes P body formation. | https://pubmed.ncbi.nlm.nih.gov/37370689/ |
| Stress granules | Renal cancer|Drug resistance | Sorafenib treatment induces formation of stress granules, stress granule-dependent stabilization of COX-2 mRNA and upregulation of COX-2 protein in renal cancer cells. | https://pubmed.ncbi.nlm.nih.gov/37216166/ |
| Super-enhancer transcriptional condensates | Castration-resistant prostate cancer | LSD1 is enriched at super enhacer transcriptional condensates and drives castration-resistant prostate cancer (CRPC) progression by activating super-enhancer-mediated oncogenic programs. Inhibitors of LSD1 and BET can synergistically suppress CRPS tumor growth in vitro in 22RV1 cells and in vivo, in mouse xenograft tumors. LSD1 inhibition represses MYC signaling in various CRPC xenograft models. | https://pubmed.ncbi.nlm.nih.gov/36877164/ |
| Androgen receptor transcriptional condensates | Castration-resistant prostate cancer|Drug resistance | Androgen receptor (AR) forms transcriptionally-active condensates in the nucleus of LNCaP and VCaP cells. Stably expressed AR-mEGFP in LNCaP and VCaP cells, as well as LNCaPcells expressing endogenously-tagged AR, form nuclear condensates upon stimulation with dihydrotestosterone (DHT). Fluorescently tagged AR recovered within seconds in FRAP experiements. AR phase separation is predominantly mediated by the NTD, although all domains contribute. Mutant-AR associated with castration-resistant prostate cancer (CRPC) forms trascriptional condensates form in response to bicalutimide AR inhibitor and are less responsive to ligand-induced regulation. ET516 is an AR LLPS inhibitor, identified in a condensate-based screen. Dissolution of mutant AR nuclear condensates upon treatment with ET516 results in reduced AR transcriptional activity in LNCaP cells, cell proliferation in LNCaP & VCaP cells, and growth of CRPC tumors in a xenograft mouse model. | https://pubmed.ncbi.nlm.nih.gov/36229685/ |
| Stress granules | Triple-negative breast cancer (TNBC)|Drug resistance | CircRNA-CREIT is downregulated in triple-negative breast cancer (TNBC) patient tissue, and was further downregulated in chemotherapy-resistant breast cancer tissues and doxorubicine (DOX) resistant TNBC cancer cell line MDA-MB-231/DOXR. Doxorubicin induces stress granules (SG) to effect drug resistance. CircRNA-CREIT attenuates SG formation via the PKR/eIF2α axis and restores DOX sensitivity in MDA-MB-231 cells as measured by colony forming assays. | https://pubmed.ncbi.nlm.nih.gov/36038948/ |
| Stress granules | Non-small cell lung cancer (NSCLC); drug resistance | three subtypes of NSCLC were identified based on the expression levels of Stress Granule Regulators. | https://pubmed.ncbi.nlm.nih.gov/35433677/ |
| Core regulatory circuitry condensates | Osteosarcoma|Drug resistance | Core regulatory circuitry factors (HOXB8 and FOSL1) form nuclear condensates in 143B cells. Recombinant intrinsically disordered regions (IDRs) of HOXB8 and FOSL1 form in vitro homotypic and heterotypic droplets. Disruption of CRC condensation by HOXB8 knock-down decreases chromatin sccessibility at superenhancer (SE) loci in 143B cells, and impairs RNA Pol II elongation on SE-driven genes, as measured by ATAC-, ChIP- and RNA-seq. Knock-down of HOXB8 causes a reduction in osteosarcoma tumor growth and metastasis in vivo, in a mouse model. Pharmacological inhibition with the H3K27 demethylase inhibitor, GSK-J4, of the CRC phase separation results in metastasis suppression and re-sensitivity to chemotherapy drugs in patient-derived xenograft model. | https://pubmed.ncbi.nlm.nih.gov/34432948/ |
| Stress granules | Frontotemporal dementia (FTD) | ELAVL4 binds MAPT RNA and co-localizes with cytosolic stress granules (SGs) in Tau-V337M neurons. EVLAVL4 gene expression is increased in organoids expressing Tau-V337M mutant, and synaptic genes are aberrantly spliced. Tau-V337M organoids exhibit early neuronal maturation, upregulation of synaptic signaling pathways and loss of deep- and upper-layer glutamatergic neurons. | https://pubmed.ncbi.nlm.nih.gov/34314701/ |
| Nuclear speckles | Alzheimer's disease (AD) | Hyaluronic acid synthase 1 (HAS1) relocalizes from cytoplasm to nuclear speckles in neurons, to reduce the synthesis and release of HA, and dysregulation of transcriptional programming. Levels of HAS1 are reduced in Alzheimer's disease and AbPP/PS1 mice. | https://pubmed.ncbi.nlm.nih.gov/38518923/ |
| P bodies | Influenza A virus (IAV) infection | Viral non-structural protein NS1 from Influenza A virus (IAV) associates with XRN1 in P bodies to promote viral replication in infected A549 cells and inhibit host immune response, as measured by RT-qPCR mRNA levels of IFN-b, IFIT1, IFIT3, IRF3, IRF7, NF-kB, ISG15. | https://pubmed.ncbi.nlm.nih.gov/34311580/ |
| TET1 nuclear bodies | Colorectal cancer | TET1 and its demethylation mark (5hmC) localize to nuclear puncta in the colorectal cancer cell lines HCT116, Caco-2 and HT-29, while in the non-transformed cell line NCM460 it localizes to the nucleus and perinuclear region. TET1 puncta partially co-localize with Cajal bodies. | https://pubmed.ncbi.nlm.nih.gov/38583183/ |
| Purinosomes | Melanoma | Stresses that increase purine demand upregulate ASB11 by epigenetic mechanisms, in HeLa and A375 cells. ASB11 promotes PAICS ubiquitination to recruit the ubiquitin-binding protein UBAP2, which governs multivalent interactions to drive purinosome assembly, in HeLa cells. Purinosome formation enhances the de novo purine synnthesis (DNPS) pathway flux. UBAP2Δ130-621 form in vitro droplets in the presence of ubiquitination reaction mix. The droplets recover after photobleaching (FRAP) on the timescale of hundreds of seconds. Melanoma A375 cells form purinosome constitutively to support their proliferation and survival. In vivo, ASB11 depletion, replacement of PAICS WT with K74R mutant, or UBAP2 depletion each retarded the growth of A375-derived tumors in xenograft mice. | https://pubmed.ncbi.nlm.nih.gov/37848033/ |
| Transcriptional condensates | Human pappilomavirus (HPV) infection|Cervical cancer | HPV18 RNAs induced transcriptional condensate at chromosome 8q24.21 in HeLa cells, recruits host transcriptional machinery in HeLa, HCT116 and HEK293T cells, and produces a HPV18/URC chimera RNA to promote cell proliferation in HeLa cells. | https://pubmed.ncbi.nlm.nih.gov/38715205/ |
| Paraspeckles | Atherosclerosis | NEAT1 -- Nuclear paraspeckle assembly transcript 1 -- is upregulated in aortas of atherosclerosis (AS) mice and dedifferentiated primary vascular smooth muscle cells (pVSMCs). Overexpression of NEAT1 induced by adenovirus enhances the proliferation and migration of dedifferentiated primary VSMCs. The proliferation and migration are mediated by NEAT1 through a mechanism involving EZH2; NEAT1 scaffolds EZH2 to epigenetically suppress downstream senescence- and antimigration-related genes in VSMCs. NEAT1 modulates the osteogenic differentiation of VSMCs in vitro and in vivo. | https://pubmed.ncbi.nlm.nih.gov/38646788/ |
| Stress granules | X-linked Charcot-Marie-Tooth Disease | Four genetic mutations in GJB1 were identified in patients suffering from X-linked Charcot-Marie-Tooth (CMT) disease. All mutants were overexpressed relative to the wild type, and are more prone to aggregation. Mutant GJB1 variants, overexpressed in HeLa cells accumulate into cytoplasmic granules that co-localize with ER markers. Expression of the CMT mutant variants of GJB1 triggers stress granule formation and impaired HeLa cell proliferation. | https://pubmed.ncbi.nlm.nih.gov/36225735/ |
| GJB1 granules | X-linked Charcot-Marie-Tooth Disease | Mutant GJB1 variants, overexpressed in HeLa cells accumulate into cytoplasmic granules that co-localize with ER markers. Expression of the CMT mutant variants of GJB1 triggers stress granule formation and impaired HeLa cell proliferation. | https://pubmed.ncbi.nlm.nih.gov/36225735/ |
| Stress granules | Amyotrophic lateral sclerosis (ALS); Frontotemporal dementia (FTD) | DHX30 is recruited into FUS cytoplasmic puncta in ALS patient spinal motor neurons, causing mitochrondrial dysfunction and cytotoxicity | DHX30 is recruited into FUS cytoplasmic puncta in ALS patient spinal motor neurons, causing mitochrondrial dysfunction and cytotoxicity |
| CAPRIN1 granules | Ataxia | Mutations in CAPRIN1 cause accumulation in cytoplasmic puncta and colocalize with other neurodegenerative disorder protein markers, including ataxia-related proteins and cause reduced neuronal activity in iPSC-derived neurons, and impaired stress granules dynamics | https://pubmed.ncbi.nlm.nih.gov/36136249/ |
| Stress granules | Rabies virus (RABV) | Stress granule (SG) induction by infection of 293T cells with the avirulent Rabies virus strain Ni-CE; the more virulent strains Nishigahara and street RABV, show reduced induction of SGs. Supression of SG induction is dependent on the sequence of the viral RABV matrix protein (RABV-M), primarily at positions 29 and 95. Variability at position 95 in RABV-M leads to phosphorylation of eiF2a in Ni-CE infected cells, but not cells infected with the more virulent strains. Cells infected with virus expressing RABV-M capable of inducing SGs stimulates RIG-I accumulation and initiation of IFN-b transcription. | https://pubmed.ncbi.nlm.nih.gov/36069552/ |
| P bodies | Epstein-Barr virus (EBV) | BGLF2 is an EBV tegument protein expressed during the EBV lytic infection stage. BGLF2 expressed in 293T cells interacts with componnets of the RISC complex. Expression of BGLF2 in HONE-1 and HeLas cells inhibited assembly of P bodies. Expression of BGLF2 inhibits let-7a miRNA function and causes an increase in SUMO proteins. | https://pubmed.ncbi.nlm.nih.gov/35007297/ |
| Stress granules | Epstein-Barr virus (EBV) infection | BGLF2 is an EBV tegument protein expressed during the EBV lytic infection stage. BGLF2 expressed in 293T cells interacts with componnets of the RISC complex. Expression of BGLF2 in HONE-1 and HeLas cells induced stress granule (SG) formation, and co-localization of BGLF2 and RISC subunits within SGs. Expression of BGLF2 inhibits let-7a miRNA function and causes an increase in SUMO proteins. | https://pubmed.ncbi.nlm.nih.gov/35007297/ |
| Insulin receptor clusters | Type 2 diabetes | Reduced accumulation of insulin receptor in insulin-resistant cells | https://pubmed.ncbi.nlm.nih.gov/36473871/ |
| Viral replication condensate | Murine norovirus (MNV) | Murine norovirus (MNV) replication leads to gradual shut-off of translation and activation of eIF2α signaling in infected RAW264.7 cell line and primary mouse bone marrow derived macrophages (BMDMs). The translation shut-off is uncoupled from the P-eIF2α signaling in MNV-infected RAW264.7 cell line, and is dependent on GCN2 activity. MNV infection causes assembly of non-canonical G3BP1-positive stress granules, that are insensitive to cyclohexamide treatment; G3BP1 co-localizes with the viral replication complexes and exhibits an altered interaction network compared to that in canonical stress granules. | https://pubmed.ncbi.nlm.nih.gov/31905230/ |
| Viral replication condensate | Zika virus (ZIKV) infection | G3BP1 and HuR relocalize into Zika virus (ZIKV) replication condensates to evade cellular stress response and promote viral replication | https://www.ncbi.nlm.nih.gov/pubmed/30944179 |
| Cajal body | Distal motor neuropathy | The homozygous point mutation (Y213H) was identified in a patient suffering from hereditary neuropathy. The mutation is located in the activation loop of VRK1 and inhibits kinase activity in vitro. Expression of mutant VRK1 in HeLa cells inhibited Cajal body assembly. | https://pubmed.ncbi.nlm.nih.gov/32365420/ |
| Non-paraspeckle NONO condensates | Cardiac fibrosis | Expression of VGLL3 gene is induced by substrate stiffness, in fibrotic heart and is specifically expressed in myofibroblasts. Overexpression of VGLL3 in myofibroblasts forms nuclear condensates, that recover from photobleaching (FRAP) on the timescale of tens of seconds. Phase separation is mediated by the glutamic acid rich low complexity domain of VGLL3. Nuclear translocation of VGLL3 in response to mechanical triggers, co-localizes with EWSR1 into non-paraspecke NONO condensates and promotes fibrosis-related gene expression in myofibroblasts. EWSR1/VGLL3 co-localization to NONO condensates increases collagen expression through inhibition of miR-29b production in cardiac myofibroblasts. | https://pubmed.ncbi.nlm.nih.gov/36754961/ |
| IRF-I transcriptional condensates | Viral infection|Age-related immuno-senescence | Ectopically-expressed IRF3 re-localizes from the cytoplasm to nuclear condensates in U2OS and HeLa cells upon infection with Sendai Virus (SeV). The nuclear condensates recover from photobleaching on the tens of seconds timescale. Recombinant GFP-IRF3 forms liquid-like droplets in vitro, that are concentration, temperature, pH and salt dependent. The liquid-like properties are demonstrated via live imaging, showing droplet fusion. Fluorescence recovery after photobleaching (FRAP) shows recovery on the tens of seconds timescale, and the mobile fraction decreases with the decrease in temperature. Phosphorylation of IRF3 at Ser386 or Ser396 promotes condesation in vitro. Condensation is mediated by the DNA-binding domain (DBD), intrinsically disordered region (IDR) and IRF-associated domain (IAD). IRF7 forms homotypic in vitro droplets, as well as heterotypic droplets with IRF3, with the heterotypic droplets exhibiting a higher mobile fraction compared to the homotypic ones. IRF3 copartmentalizes IRF7 in the nucleus of infected U2OS infected with SeV. SIRT1 is required for IRF3 condensation and transcription of inflammatory genes (e.g., IFNB1) in SeV-infected HEK293T cells. Myeloid SIRT1 deficiency inhibits IRF3 and IRF7 nuclear condensate formation and causes inate immune evasion in mice infected with SeV, VSV and HSV-1. Mechanistically, IRF3/IRF7 condensation is inhibited by site specific acetylation within the DBD; deacetylation by SIRT1 promotes condensation in vitro and in infected cells. | https://pubmed.ncbi.nlm.nih.gov/35879450/ |
| SFPQ condensates | Cancer | SFPQ undergoes phase separation through its prion-like domain to form nuclear condensates in HCC and Hep3B cells. The condensates recover after photbleaching (FRAP) on the timescale of tens of seconds. SFPQ, restrains Smad4 in its condensates and triggers dissociation of the Smad activator complex. SFPQ suppresses TGF-b-induced growth-inhibitory and transcriptional responses in several cell lines. SFPQ deficiency or loss of LLPS ability attenuates tumorigenesis. Nuclear SFPQ condensates driven by SFPQ overexpression suppress TGF-β tumor suppression signaling in cultured cells and in vivo, in xenograft mice. | https://pubmed.ncbi.nlm.nih.gov/38103553/ |
| RIα condensates | Fibrolamellar carcinoma (FLC) | Endogenous PKA regulatory subunit RIα forms cytoplasmic condensates in HEK293A cells, that recover after photbleaching on the tens of seconds timescale. Recombinant RIα forms in vitro droplets. cAMP promotes condensation of RIα both in vitro and in cells. RIα condensates dynamically sequester cAMP and retain high PKA activity. RIα phase separation is necessary for effective cAMPcompartmentation. A PKA oncoprotein disrupts RIα bodies, leading to aberrant signaling and growth. DnaJB1-PKA cat fusion oncogene is expressed in fibromellar carcinoma (FLC) and disrupts RIα condensates and cAMP compartmentalization, resulting in increased cell proliferation and transformation in cellular models. | https://pubmed.ncbi.nlm.nih.gov/32846158/ |
| Stress granules | Pancreatic ductal adenocarcinoma (PDAC) | Stress granules (SGs) are upregulated in an obesity-associated pancreatic ductal adenocarcinoma (PDAC) mouse model. SG upregulation is mediated by obesity-associated IGF1 and hyperactivation of IGF1R in a panel of human and murine cells, in tumor sections, as well as in a PDAC mouse model. IGF1 promotes SG formation in PDAC cells by modulating S6K1-mediated partitioning of SRPK2 to SGs and activation of IGF1/PI3K/mTOR/S6K1 pathway. Obesity-associated PDAC is selectively dependent on S6K1 for SG formation and tumor growth in an in vivo mouse model. | https://pubmed.ncbi.nlm.nih.gov/35674408/ |
| FXR1 granules | Fragile X syndrome (FXS) | FXR1(N202S) point mutation, associated with FXS, prevented assembly of the FXR1 mRNP granule network and associated signaling. | https://www.biorxiv.org/content/10.1101/2023.11.05.565677v3 |
| Viral replication condensate | Barley yellow striate mosaic virus (BYSMV) infection | BYSMV P-protein forms cytoplasmic granules in N. benthamiana leaves. | https://pubmed.ncbi.nlm.nih.gov/35191833/ |
| Glycolytic bodies (G-bodies) | Hepatitis; cirrhosis; hepatocellular carcinoma (HCC) | PFKL, AMPK, RPIA and PKM2 localize to G-bodies, whose size increase with liver disease progression, from chronic hepatitis, to active hepatitis, cirrhosis and HCC. | https://www.biorxiv.org/content/10.1101/2024.02.29.582877v1 |
| Stress granules | Porcine deltacoronavirus (PDCoV) infection | Infection with porcine deltacoronavirus (PDCoV) triggered induction of G3BP1-positive stress granules in LLC-PK1 cells, in a PERK-dependent manner. Overexpression of G3BP1 inhibited PDCoV replication, while G3BP1 knockdown promoted viral replication. G3BP1 phosphorylation enhanced PDCoV-induced NF-κB subunit p65 phosphorylation and inflammatory cytokine production. | https://pubmed.ncbi.nlm.nih.gov/38593624/ |
| Glycolytic bodies (G-bodies) | Malaria infection | The malaria parasite, Plasmodium falciparum, forms PFK9-containing G-body-like condensates as an adaptive mechanism to host blood sugar levels. | https://www.biorxiv.org/content/10.1101/2024.04.08.588540v3 |
| FMRpolyG / CGG repeat foci | Fragile X-related tremor/ataxia syndrome (FXTAS) | CGG repeat RNA-G quadruplexes colocalize with the RAN-translation product - FMRpolyG peptides in Neuro-2a cells and in vitro reconstituted droplets. FMRpolyG forms homotypic in vitro droplets that recover from photobleaching (FRAP) on the timescale of minutes; heterotypic FMRpolyG:(CGG)99 droplets have reduced mobile fracton compared to the FMRpolyG homotypic droplets. FMRpolyG forms a complex with RNA-binding proteins and exosomal proteins, mediated by RNA interactions in hypocampus of CGG-knock-in mice. FMRpolyG can propagate via exosomes, eliciting neuronal dysfunction, as measured by amplitude and frequency of spontaneous excitatory postsynaptic currents and firing frequency in primary neurons from CGG-knock-in mice. | https://pubmed.ncbi.nlm.nih.gov/33523882/ |
| FMRpolyG foci | Fragile X-related tremor/ataxia syndrome (FXTAS) | FMRpolyG foci are detected, co-localized with Ubiquitin, in hippocampal brain sections of Fragile X-related tremor/ataxia syndrome (FXTAS) patients, but not in age-mathced control individuals. Translation of expanded CGG repeats in mice leads to accumulation of FMRpolyG and Ubiquitin foci in the brain, and FXTAS-like pathogenicity as measured via behavioral tests (e.g., locomotor deficiency, increased falling from rotarod, decreased tracion ability, decreased grip strength and decreased number of rears in open field observation), development of obesity and early death. Mechanistically, FMRpolyG foci trap LAP2β and cause nuclear lamina disorganization in N2A cells, mouse hippocampus and patient brain sections. | https://pubmed.ncbi.nlm.nih.gov/28065649/ |
| CGG repeat foci | Fragile X-related tremor/ataxia syndrome (FXTAS) | Expanded CGG repeats form nuclear RNA foci in multiple cultured cell models. CGG repeat foci sequester Sam68, hnRNP-G and MBNL1 and inhibit Sam68-dependent alternative splicing in cultured cells. Alternative splicing is also altered in FXTAS patients. Sam68 co-localizes with CGG aggregates in brain sections from mouse models and FXTAS patients. Fluorescence recovery after photobleaching (FRAP) of Sam68 nuclear condensates in COS7 cells shows a decrease in the mobile fraction in cells that also express CGG repeats. Recovery occurs on the minutes timescale. | https://pubmed.ncbi.nlm.nih.gov/20186122/ |
| Viral replication condensate | Chandipura virus (CHPV) infection | Heterogenoeus inclusion bodies composed of CHPV viral proteins and host stress granule proteins to evade formation of antiviral stress granules and and promote viral replication. | https://www.biorxiv.org/content/10.1101/2023.12.13.571494v1 |
| NTRK-fusion condensates | Cancer | NRTK fusion oncoproteins form liquid-like cytoplasmic condensates in HEK293 cells stably expressing the respective fusion. NRTK fusion condensates fuse and recover from photobleaching on the 10s of seconds timeframe. NRTK fusion condensates, accelerate kinase reactions and promote downstream signaling, such as TRK activation. | https://pubmed.ncbi.nlm.nih.gov/37812694/ |
| ELM4-ALK condensates | Non-small cell lung cancer (NSCLC) | Endogenous expression of fusion oncogenes ELM4-ALK V1 and ELM4-ALK V3 in non-small cell lung cancer cell lines H3122 and H2228, respectcively, as well as overexpression in HEK293 and inducible Beas2B cells leads to formation of cytoplasmic condensates. Downstream signaling proteins, such as GRB2, SOS1, c-KIT, PI3K and PLCγ2 colocalize in the ELM-ALK cytoplasmic puncta in H3122, H2228 and inducible Beas2B cells. Foci formation is dependent on an active kinase conformation and is driven by dimerization, with a requirement for stoichiometric mismatch between ELM4 and ALK proteins. | https://pubmed.ncbi.nlm.nih.gov/34661367/ |
| CPSF6 foci | Colorectal cancer | CPSF6 is expressed at higher levels in colorectal cancer (CRC) cell lines SW480 and RKO, relative to the untransformed cell line NCM460. The CRC cell lines form reduced CPSF6 nuclear condensates, which is associated with the presence of shorter 3' UTRs. CPSF5 localizes to CPSF6 condensates in NCM460 and HEK293 cells. Recobinant GFP-CPSF6-IDR forms homotypic droplets in vitro, that fuse and recover from photobleaching (FRAP) on the tens of seconds timescale. Endogenously tagged and overexpressed GFP-CPSF6, as well as overexpressed GFP-CPSF6-IDR nuclear condensates recover from FRAP on the tens of seconds timescale. CLK2 modulates alternative polyadenylation and cell proliferation by regulating CPSF6 condensation. | https://pubmed.ncbi.nlm.nih.gov/37777964/ |
| Stress granules | Colorectal cancer (CRC)|Drug resistance | YTHDC2 expression is donwregulated in 5-FU resistant colorectal cancer (CRC) cell line SW480. Overexpression of YTHDC2 increased the sensitivity of HCT116 cells to 5-FU, and knockdown of YTHDC2 increased the IC50 value of 5-FU in SW480 cells. YTHDC2 overexpression increased the apoptosis rate and cell cycle arrest in G0/G1 of HCT116 cells treated with 5-FU, while knockdown of YTHDC2 significantly decreased the apoptosis rate of SW480 cells. YTHDC2 knockdown increased the proliferation of SW480 cells, and in vivo decreased apoptosis and increased proliferation in 5-FU treated xenograft mouse models with YTHDC2 knockdown. eIF2α phosphorylation promotes stress granule (SG) formation, and the LIMK1/eIF2α regulatory axis attenuates 5-FU-induced apoptosis, triggering chemoresistance in CRC cellular and mouse models. YTHDC2 regulates LIMK1 expression through m6A modification and LIMK1 reduces the 5-FU-induced apoptosis via activating ER stress. | https://pubmed.ncbi.nlm.nih.gov/37778684/ |
| Viral replication condensate | Tombusvirus (TBSV)|Carnation Italian ringspot virus | Core autophagy proteins (ATG8, ATG5, ATG101, ATG4 and ATG1a) are recruited into viral replication organelles (VROs), cosisting of clustered peroxisomes in N. benthamiana leaves upon infection with tombusvirus (TBSV) and carnation Italian ringspot virus (CIRV)and localize with viral replication proteins p33 and p36, respectively. Functionally, the recruitment of autophagy proteins to VROs promotes TBSV and CIRV viral replication and inhibits autophagic flux in plants. NBR1 and ATG8 in VROs recover from photobleaching (FRAP) on the minutes timescale. | https://pubmed.ncbi.nlm.nih.gov/38484009/ |
| Cajal body | Colorectal cancer (CRC)|Drug resistance | Cajal body number and morphology differ between a 5-FU resistant and a sensitive colorectal cancer (CRC) HCT-8 cell line. Coilin phosphorylation is increased in 5-FU resistant HCT-8/FU cells relative to HCT-8 cells. UMHK1 is selectively overexpressed in 5-FU resistant HCT-8 cell lines to promote coilin phosphorylation. Cajal bodies disassembled and decreased in numbers following 5-FU treatment in HCT-8 cells. Removal of 5-FU caused Cajal body reassembly in the surviving cells, and eventually recovered to normal counts as well as morphology. UHMK1 expression was correlated with both coilin phosphorylation level and Cajal body count per cell. | https://pubmed.ncbi.nlm.nih.gov/35151311/ |
| Stress granules | Myotonic dystrophy type 1 (DM1) | MBNL1 and CUGBP1 colocalize with stress granules (SGs) in human lens epithelial cells (HLECs) exposed to oxidative stress. SGs disperse more quickly in DM1 HLECs compared to SGs in control cells. SG formation and dispersal are altered by CUGexp RNA expression in an HeLa DM1 cellular model. SG dynamics are altered by reduction in expression of MBNL1 or CUGBP1 in HeLa DM1 cell model, as measured by FRAP. SGs in HeLa cells expressing CTG expansion contain less poly(A) RNA and show altered docking events with P bodies. | https://pubmed.ncbi.nlm.nih.gov/35642886/ |
| Stress granules | Porcine reproductive and respiratory syndrome virus (PRRSV) infection | Viral replicase Nsp1β is sorted into porcine reproductive and respiratory syndrome virus (PRRSV)-induced stress granules (SGs) during infection in MARC-145 cells, where it co-localizes with PKR. G3BP1 promotes the nsp1β-PKR interaction and contributes to nsp1β-mediated inhibition of PKR activity, to evade inflammatory host response. | https://pubmed.ncbi.nlm.nih.gov/35858300/ |
| Stress granules | Red-spotted grouper nervous necrosis virus (RGNNV) | Red-spotted grouper nervous necrosis virus (RGNNV) infection induced stress granule (SG) production in GS cells through a viral replication- and PERK pathway-dependent mechanism. induces assembly of stress granules. Overexpression of G3BP1 up-regulated interferon immune response and pro-inflammatory cytokines. | https://pubmed.ncbi.nlm.nih.gov/35935992/ |
| Stress granules | African swine fever (ASF) infection | Expression of Africal swine virus (ASF) protein MGF110-7L in HEK293T cells induces host ATF4 translation, phosphorylation of eIF2α, activation of integrated stress response (ISR) and host cell translation arrest. MSG110-7L triggest stress granules (SG) formation in a phosphorylated eIF2α-dependent manner in PK-15 cells. ASF virus expression induces SG formation through a eIF2α phosphorylation-dependent mechanism, by activating the PERK and PKR pathways. | https://pubmed.ncbi.nlm.nih.gov/36377947/ |
| Stress granules | Peste des petits ruminants virus (PPRV) infection | Peste des petits ruminants virus (PPRV) N protein interacts with host protein interferon-inducible double-stranded RNA-dependent protein kinase activator A (PACT) in infected HEK293T cells. PACT deficiency promotes PPRV replication in infected Vero cells. PPRV N protein induces stress granule (SG) formation in PPRV N-transfected and PPRV infeced Vero cells, and triggers PKR phosphorylation in a PACT-dependent manner. SG induction is dependent on viral replication; PPRV-induced SGs predominantly hijack host mRNA but not viral RNA that benefits PPRV multiplication. | https://pubmed.ncbi.nlm.nih.gov/36651745/ |
| Stress granules | Viral hemorrhagic septicemia virus (VHSV) | Viral Hemorrhagic Septicemia Virus (VHSV) infection of RTG2 and RTgill cells triggers stress granule formation through a PERK-, and active viral replication-dependent mechanism, and inhibit the ability of the cell to induce an antiviral response downstream of IFN production. SG induction is dependent on viral replication; G3BP1 regulates IFN signaling during VHSV infection and is required for viral protein production. | https://pubmed.ncbi.nlm.nih.gov/36851680/ |
| Double-stranded RNA-induced foci | Zika virus (ZIKV) infection; Dengue virus infection | Cytoplasmic puncta form in response to Zika and Dengue virus infection, or dsDNA and incorporate activated PKR, RNase L, and OAS3. | https://www.biorxiv.org/content/10.1101/2023.05.30.542827v1 |
| Stress granules | Respiratory syncytial virus (RSV) infection; parainfluenza virus infection | Stress granules assembly is induced by viral infections in cells containing high but not low levels of copy-back viral genomes. | https://pubmed.ncbi.nlm.nih.gov/37983241/ |
| Cajal body | Hereditary spastic paraplegia (HSP) | The kinase inactive VRK1-D263G mutant is found in HSP patients with neurologic phenotype and moderate intellectual disability. The mutation causes loss of kinase function in VRK1. When expressed in HeLa cells, the mutant inhibits assembly of Cajal bodies. | https://pubmed.ncbi.nlm.nih.gov/34504951/ |
| DNA damage foci | Hereditary spastic paraplegia (HSP) | The kinase inactive VRK1-D263G mutant is found in HSP patients with neurologic phenotype and moderate intellectual disability. The mutation causes loss of kinase function in VRK1. When expressed in HeLa cells, the mutant inhibits formation of DNA damage response bodies detected via 53BP1 foci. | https://pubmed.ncbi.nlm.nih.gov/34504951/ |
| LATS1 foci | Breast cancer | SNHG9 lncRNA is upregulated in xenografted breast tumors compared to cultured cancer cells, based on a GWAS analysis. SNHG9 expression correlates with lower survival rates for patients and YAP activation in advanced breast cancer. In colony forming assays, expression of SNHG9 promotes breast cancer cell anchorage-independent growth. SNHG9 lncRNA induces LATS1 LLPS, co-localizes with LATS1 and phosphatidic acids in cytoplasmic condensates and inhibits the kinase activity of LATS1. LATS1-GFP overexpression in HEK293A cells forms dynamic cytoplasmic puncta which recover from photobleaching on the timescale of seconds. LLPS is driven by the disordered prion-like domain (PrLD). LATS1-GFP phase separates in vitro, in droplets with FRAP recovery rates on the seconds-minutes time scale; LLPS is mediated by the PrLD domain. In vitro-transcribed SNHG9 co-localizes with LATS1 in in vitro droplets and promotes droplet formation. In xenograft mice injected with MDA-MB-231 cells, SNHG9 expression drives formation of LATS1 condensates, inhibits LATS1 kinase activity, promotes tumor growth, YAP1 expression and microvascularization index. | https://pubmed.ncbi.nlm.nih.gov/34267352/ |
| Stress granules | Orthoreovirus infection | Infection of A549 cells with reovirus induces stress granule (SG) formation, independent of PKR, HRI, PERK or GCN. Viral σ3 double-stranded RNA-binding protein inhibits SG formation and activation of PKR in infected A549 cells, promoting viral replication. A virus harboring the K287T mutation which inhibits σ3's dsRNA-binding ability, was less virulent compared to the wild-type, in vivo, in infected neonatal C57BL/6 mice,and prevented these animal from developing myocarditis. | https://pubmed.ncbi.nlm.nih.gov/34237110/ |
| Stress granules | SARS-CoV-2 | SARS-CoV-2 infection of ACE2 expressing HeLa cells inhibits stress granule (SG) formation in response to oxidative stress, to promote viral replication. SGs are formed in these cells when treated with sodium arsenite, poly(I:C) or SARS-CoV-2 RNA. G3BP1 and PKR are required for SG formation in response to SARS-CoV-2 RNA. Expression of the SARS-CoV-2 nucleocapsid protein is required for SG inhibition, through a mechanism that involves interaction with G3BP1 and PKR. This mechanism of SG suppression by the viral nucleocapsid is shared between SARS-CoV, SARS-CoV-2 and MERS-CoV viruses. | https://pubmed.ncbi.nlm.nih.gov/34035218/ |
| N-protein/IKK condensates | SARS-CoV-2 infection | SARS-CoV-2 nucleocapsid protein forms cytoplasmic condensates that recruit IKKb and TAK1 in SARS-CoV-2 infected Huh7 cells, and hyper-activate NF-kB signaling and inflammation. | https://pubmed.ncbi.nlm.nih.gov/33895773/ |
| DACT1 condensates | Bone metastasis | TGF-b mediated induction of DACT1 results in formation of cytoplasmic condensates and suppression of Wnt activation and promotes bone metastasis growth in mice. | https://pubmed.ncbi.nlm.nih.gov/33723425/ |
| Stress granules | Low-grade astrocytoma; glioblastoma (GBM) | G3BP2 and TIAR-positive tress granules (hypoxic stress responses) are activated in low grade astrocytoma and glioblastoma (GBM) tissues, and correlate with poor patient ourcomes. | https://pubmed.ncbi.nlm.nih.gov/33203845/ |
| Stress granules | Pseudorabies virus (PRV) | PRV infection inhibits oxidative stress-induced stress granule (SG) formation in infected Vero and PK-15 cells, inhibited eIF2a phosphorylation, and activated PKR and PERK kinases during the early infection stage, to facilitate virus replication. | https://pubmed.ncbi.nlm.nih.gov/32768230/ |
| Tax-speckle structures | Adult T-cell leukemia/lymphoma (ATL) | HTLV-1 viral oncoprotein Tax impairs DNA-damage response (DDR) through formation of Tax-speckle structures (TSS) containing RNF8 and K63-linked polyubiquitin chains, which sequester proteins involved in DDR (e.g., BRCA1, DNA-PK, MDC1), in HTLV-1 infected HeLa cells. | https://pubmed.ncbi.nlm.nih.gov/32453758/ |
| Tau tangles | Alzheimer's disease (AD); Pick's disease (PiD); progressive supranuclear palsy (PSP) | BRCA1 aberrantly localizes into tau cytoplasmic aggregates in Alzheimer's disease (AD), Pick's disease (PiD), and progressive supranuclear palsy (PSP) patient brains. | https://pubmed.ncbi.nlm.nih.gov/31861888/ |
| Cajal body | Spinal muscular atrophy (SMA)|pontocerebellar hypoplasia (PCH)|microcephaly|amyotrophic lateral sclerosis (ALS)|distal motor neuron dysfunctions | G135R rare mutation in VRK1 kinase destabilizes the kinase, reduces its half-life and prevents formation of Cajal bodies in HEK-293T cells. | https://pubmed.ncbi.nlm.nih.gov/31527692/ |
| 53BP1 foci | Spinal muscular atrophy (SMA)|pontocerebellar hypoplasia (PCH)|microcephaly|amyotrophic lateral sclerosis (ALS)|distal motor neuron dysfunctions | G135R rare mutation in VRK1 kinase prevents formation of 53BP1 foci induced in response to DNA-damage agents. VRK1-G135R mutant destabilizes the kinase and reduces its half-life in HEK-293T cells. | https://pubmed.ncbi.nlm.nih.gov/31527692/ |
| Stress granules | Human enterovirus D68 (EV-D68) infection | Stress granule (SG) formation is induced during early infection and inhibited in late stages EV-D68 infection in 293T cells. EV-D68 infection induces PKR and eIF2a phosphorylation-dependent SGs. Overexpression of SG proteins, G3BP1, TIA-1 or HUR, inhibit viral replication. SG proteins interact with 3' UTR of viral RNA to inhibit viral replication. Inhibition of stress SG assembly during late infection stages via cleavage of G3BP1 by 3Cpro protease. | https://pubmed.ncbi.nlm.nih.gov/31941779/ |
| Stress granules | Foot-and-mouth disease virus (FMDV) infection | FMDV infection of αvb6 cells leads to inhibition of stress granule formation (SG) under oxidative stress conditions, in a mechanism dependent on the catalytic activity of the viral proteases Lpro and 3Cpro, which cleave G3BP1 and G3BP2. | https://pubmed.ncbi.nlm.nih.gov/30404792/ |
| Stress granules | Enterovirus (EV) infection | Infection of HeLa cells with four different members of the enterovirus (EV) suppressed stress granule (SG) formation and activation of type I IFN-dependent gene transcription. These mechanisms are mediated by the EV 2Apro and require its protease activity. | https://pubmed.ncbi.nlm.nih.gov/30867299/ |
| Stress granules | Chronic myelogenous leukemia (CML) | BCR-ABL oncoprotein accumulates into cytoplasmic granules in a panel pf Ph-positive cell lines, as detected by immunofluorescence. Expression of BCR-ABL-EGFP causes formation of cytoplasmic granules in HeLa and Cos-1 cells. These cytoplasmic granules exhibit liquid-like behavior, as inferred from granules fusion observed using live imaging. In HeLa cells, BCR-ABL-EGFP and localizes to stress granules in a tyrosine kinase activity-dependent manner. BCR-ABL granule formation is require to promote Ba/F3 cell proliferation. | https://pubmed.ncbi.nlm.nih.gov/31735741/ |
| Cajal body | Distal hereditary motor neuropathies (dHMNs) | Compound heterogeneous mutations in VRK1 identified in sibling patients of Distal hereditary motor neuropathies (dHMNs). These mutations cause protein mislocalization from the nucleus to the cytoplasm and altered coilin expression in patients fibroblasts. VRK1 mutations lead to inhibition of Cajal body formation and impaired neurite length and branching in hiPSC-derived motor neurons. | https://pubmed.ncbi.nlm.nih.gov/31090908/ |
| Stress granules | Alzheimer's disease (AD) | 5XFAD AD mouse models subjected to anesthesia and surgery exhibit stress granule induction and localization of SYK and phosphorylated SYK within stress granules. | https://pubmed.ncbi.nlm.nih.gov/34512311/ |
| P bodies | Cancer | P body assembly is inhibited by highly phosphorylated EDC3 in tumor cells to promote tumor growth and invasion. | https://pubmed.ncbi.nlm.nih.gov/33586867/ |
| Stress granules | Breast cancer | Stress granules formation is accelerated in breast cancer patients with low levels of PTEN expression and is associated with poor progrnosis and antitumor immune response and drug resistance. | https://pubmed.ncbi.nlm.nih.gov/39080255/ |
| ORC1 bodies | Human immunodeficiency virus type 1 (HIV-1) | ORC1 is a host factor that associates with the HIV-1 promoter and host repressive epigenetic factors within nuclear condensates to maintain enriched repressive histone modifications (HDAC1, HDAC2) on the HIV-1 promoter, as a mechanism for maintaining HIV-1 latency in J.Lat cells. ORC1 regulates DNA methylation of the HIV-1 promoter, through interactions with DNMT1. ORC1 form nuclear condensates and co-localizes with HP1a, SUV39H1, EZH2, H3K9me3, H3K27me3 in ORC1-GFP transfected HEK293 cells. The ORC1 nuclear condensates recover from photobleaching (FRAP) on the timescale of tens of seconds, and exhibit fusion and fission, demonstrating liquid-like fluidity. GFP-ORC1-IDR forms condensates in vitro in a concentration-dependent fashion; the in vitro condensates dissolve with increase in buffer ionic strength. Depletion of ORC1 reactivated HIV-1 in primary CD4+ T-cells from people living with HIV-1. | https://pubmed.ncbi.nlm.nih.gov/39082875/ |
| FOXK1 condensates | Chronic kidney disease (CKD) | Aberrant, nuclear FOXK1 transcriptional condensates form in fibrotic renal tubular epithelial cells with upregulated levels of FOXK1. These condensates boost glycolysis through upregulation of transcription of glycolytic-related genes to promote relan fibrosis. | https://pubmed.ncbi.nlm.nih.gov/39083268/ |
| Chandipura virus inclusion bodies | Chandipura virus (CHPV) | CHPV proteins nucleocapsid (N), phosphoprotein (P) and large (L) form cytoplasmic inclusion bodies (CHPV-IBs) and recruit host stress granule proteins (TIA-1, PABP1, eIFη, G3BP1, Ago2) in CHPV infected Vero and N2A cells. CHPV-IB form independently of PKR and eIF2a phosphorylation, but recruit PKR to promote viral replication. | https://pubmed.ncbi.nlm.nih.gov/39066190/ |
| Paraspeckles | Karposi's Sarcoma herpresvirus (KSHV) infection | KSHV ORF11 protein induces SFPQ-positive, paraspeckle-like nuclear puncta during early stages of lytic replication, that contribute to viral and host RNA processing, promote genome instability and viral replication. | https://www.biorxiv.org/content/10.1101/2024.07.23.604779v1 |
| P bodies | Colorectal cancer (CRC) | YAP expression increases the number of P bodies in cancer cells, in a mechanism dependent on YAP/TEAD transcriptional activity, to promote cell proliferation and migration. | https://pubmed.ncbi.nlm.nih.gov/39046443/ |
| Nucleolus | Hendra virus (HeV) infection | HeV protein M accumulates in the nucleolar FC-DFC subcompartments during infection and inhibits host ribosomal RNA synthesis and nucleolar DNA damage repair pathway. | https://www.biorxiv.org/content/10.1101/2023.08.10.552741v2 |
| WTAP condensates | Inflammatory disease | WTAP expression is upregulated in patinets suffering from systemic lupus erythematosus, astma, sepsis, rheumatoid arthritis, psoriasis and Crohn's disease. WTAP levels are elevated in mouse lung tissue with LPS-induced lung injury, and THP-1 cells and macrophages exposed to multiple inflammatory stimuli. WTAP positively regulates proinflamatory responses in THP-1 and 293T cells, and aggravates LPS-induced sepsis in mice. WTAP undergoes phase separation to promote METTL3 rectuitment and expression of proinflammatory genes through m6A modification. WTAP forms condensates in vitro, that recover from photobleaching (FRAP) on the tens of seconds time scale. | https://pubmed.ncbi.nlm.nih.gov/39007267/ |
| NAT10 condensates | Gastric cancer | NAT10 expression is elevated in gastric patient samples and correlates with unfavorable prognosis, advanced state disease and metastasis. NAT10 forms condensates in gastric cancer cells to promote cancer progression, migration and invation, supported growth of patient-derived organoids, and accelerated tumor growth. The C-terminal disordered region-mediated LLPS if NAT10 was required for tumor promotic function in gastric cancer cells. Oncogenesis is mediated by a molecular mechanism where NAT10 LLPS stimulates acetylation of SRSF2, which modulates splicing of YTHDF1. | https://pubmed.ncbi.nlm.nih.gov/39024555/ |
| KAT6A condensates | Ovarian cancer; drug resistance | Increased number of KAT6A nuclear condensates is observed in PARPi resistant ovarian cancer cells, to inhibit PARPi-induced PARP1 bound to chromatin, enhance DNA damage repair under PARPi treatment and support tumor growth. | https://pubmed.ncbi.nlm.nih.gov/38973255/ |
| Transcriptional condensates | Lung adenocarcinoma (LUAD) | SP1 expression levels are higher in LUAD tissues compared to normal lung tissue. SP1 forms condensates at the superenhancer site that controls RGS20 transcription to promote cell proliferation and metastasis. | https://pubmed.ncbi.nlm.nih.gov/38976739/ |
| Stress granules | Zika virus (ZIKV) infection | Zika virus strain GUINEA-18 suppresses stress granule formation in response to osmitoc and oxidative stress, to promote lower viral replication rates, innate immune response and weaker cytotoxicity compared to a strain that does not suppress stress granule formation, in infected A549 cells. | https://pubmed.ncbi.nlm.nih.gov/38968296/ |
| Germ granules | Cancer | Cytoplasmic DDX4-positive, germ-like granules are observed in the cytoplasm of several cancer patient tissues, but not in the normal tissue. The aberrant condensates are further detected in xenograft tumors and spheroid models. DDX4 aberrant condensates are associated with transcriptional reprogramming, tumor growth, cancer invasiveness and poor patient survival. | https://pubmed.ncbi.nlm.nih.gov/38963760/ |
| Stress granules | Zika virus (ZIKV) | ZIKV protein NS2B promotes eIF2α dephosphorylation through a PP1α-dependent mechanism, to inhibit stress granule (SG) formation under oxidative stress, and evade the host antiviral response in infected HeLa cells. The NS2B-mediated mechanism of SG suppression is shared among multiple members of the flavivirus family: Dengue, West Nile, Yellow Fever, Zika and Japanese encephalitis virus. | https://pubmed.ncbi.nlm.nih.gov/38935808/ |
| Stress granules | Myocardial inflammation | Stress granule levels are reduced in vivo, in a diabetes ischemia-reperfusion rat model, and in a high glucose hypoxia/reoxygenation cell model and correlate with increased myocardial tissue damage, apoptosis and inflammatory factors. | https://pubmed.ncbi.nlm.nih.gov/38988216/ |
| Stress granules | Amyotrophic lateral sclerosis (ALS); Frontotemporal dementia (FTD) | Cytoplasmic TDP-43 inclusions co-localize with stress granules markers in cellular models and ALS and FTLD-U patient brain tissue. Cytoplasmic inclusions of TDP-43 disease associatred mutations correlate with increase in insoluble protein aggregates and cellular toxicity. | https://pubmed.ncbi.nlm.nih.gov/20948999/ |
| P bodies | Trypanosoma cruzi (T.cruzi) infection | T. cruzi infection induces P body assembly in HT1080 cell line, to supress the host immune response by inhibiting IL-1b production. | https://pubmed.ncbi.nlm.nih.gov/39122157/ |
| WNK bodies | Hypokalimia | WNK bodies assemble in the cytoplasm in response to hypokalemia and function as signal amplifiers that mediate tubular potassium responsiveness, nephron sexual dimorphism, and blood pressure salt-sensitivity | https://www.biorxiv.org/content/10.1101/2021.03.12.435046v2 |
| CRYAB bodies | Ischemic cardiomyopathy (ICM) | CRYAB phosphorylated at Ser59 accumulates in NP-40 resistance bodies in ICM patient heart tissue and in cardiac ischemia-reperfusion injury mouse models to cause desmin and actin mislocalization and left ventricular systolic dysfunction post-myocardial infarction | https://www.biorxiv.org/content/10.1101/2024.08.30.610556v1 |
| DAZAP1 nuclear condensates | Oral squamous cell carcinoma (OSCC) | Overexpression of DAZAP1 is associated with nuclear induction of DAZAP1 condensates, increase COX16 expression and mitochondrial metabolism, and promoted OSCC invasion, as well as poor patient prognosis. | https://pubmed.ncbi.nlm.nih.gov/39120588/ |
| Nucleolus | Ribosomopathy | Mutations in SNORD118, found in patients suffering from ribosomapaties with CNS-exclusive pathology, cause aberrant nucleolar morphology and impaired ribosome biogenesis in iPSC-derived neural progenitor cells and inhibit cerebral organoid growth. | https://pubmed.ncbi.nlm.nih.gov/39108718/ |
| Stress granules | Ataxia; Amyotrophic lateral sclerosis (ALS); Frontotemporal dementia (FTD) | Inhibition of stress granule formation exacerbates disease phenotypes in Drosophila models of neurodegeneration and in cultured primary neurons. | https://pubmed.ncbi.nlm.nih.gov/39106168/ |
| ARID1A nuclear condensates | Ewing's sarcoma; Osteosarcoma | ARID1A is overexpressed in Ewing's sarcoma cancer lines, forms nuclear condensates at EWS/FLI1 sites, upregulates expression of cancer-related genes, promotes tumor growth in xenograft mice and spheroid invasion. | https://pubmed.ncbi.nlm.nih.gov/39095374/ |
| P bodies | Acute myeloid leukemia (AML) | Elevated levels of P-bodies that sequester mRNA encoding for tumor suppressors promote leukemia progression in vivo in AML mouse models. | https://pubmed.ncbi.nlm.nih.gov/39169219/ |
| sfRNA granules|P bodies|RNase L-induced bodies | Zika virus (ZIKV)|West Nile virus (WNV)|Dengue virus (DENV) | Subgenomic flavivirus RNAs (sfRNAs) bodies associate with P bodies upon infection - viral RNA replication phase, where they are exposed to the host RNA decay machinery; activation of RNase L as part of the host antiviral response they relocalize to RNase L-induced bodies to drive viral RNA decay. | https://pubmed.ncbi.nlm.nih.gov/39196777/ |
| Nuclear speckles | Epileptic seizures | Nuclear speckles in dentate gyrus neurons - monitored via SRSF2 immunofluorescence, undergo morphological changes, including size and share distribution, upon neuronal activation in rat models of epileptic seizures. These morphological changes correlate with changes in transcriptional and splicing programming and protein composition within the nuclear speckles. | https://pubmed.ncbi.nlm.nih.gov/39197592/ |
| Stress granules | Cancer | Environmental exposure to toxic heavy metal Cr(VI) induces stress granule formation to promote malignant properties, such as proliferation rate, sphere formation and malignant markers. | https://pubmed.ncbi.nlm.nih.gov/39306064/ |
| phospho-HDAC6 condensates | Triple-negative breast cancer (TNBC) | Nuclear phospho-HDAC6 is overexpressed in TNBC cells, forms nuclear condensates to alter chromatin structure, promote tumor growth in xenograft models and correlates with the tumor stage. | https://pubmed.ncbi.nlm.nih.gov/39198689/ |
| YTHDF2 condensates | Diabetic cardiac fibrosis | Expression of YTHDF2 is increased in mouse fibrotic heart tissue, diabetic model of cardiac fibroblasts, and fibrotic heart tissue from diabetic cardiomyopathy. YTHDF2 abnormally accumulates into condensates, leading to increase in expression of fibrotic markers, enhanced mitochondrial function and impairment of heart function, such as reduced fractional shortening and decreased ejection fraction. | https://pubmed.ncbi.nlm.nih.gov/39342271/ |
| Stress granules | Neurodegeneration | Dipeptide repeats poly-PR and poly-GR, formed through translation of C9ORF72 repeat expansion, induce formation of and localize to stress granules, to impair translation. | https://pubmed.ncbi.nlm.nih.gov/27768896/ |
| NUP98-HOXA9 condensates | Acute myeloid leukemia (AML) | NUP98-HOXA9 form nuclear puncta in HEK293T cells under overexpression and in primary mouse hematopietic stem cells. Expression of NUP98-HOXA9 in primary hematopoietic stem cells causes aberrrant expression of HOX family genes and oncogenic transformation. NUP98-HOXA9 nuclear condensates formed by overexpression in HEK293T cells recover after photobleaching (FRAP) in seconds. In vitro formed NUP98-HOXA9 condensates age rapidly. LLPS behavior is regulated by FG-motif interactions and HOXA9 domain-DNA interactions. | https://pubmed.ncbi.nlm.nih.gov/34903620/ |
| Viral replication condensate | Gastroenteritis; Norovirus infection | Human norovirus (HuNoV) RNA-dependent-RNA polymerase (RdRp) form condensates with enzymatically active RdRp. Pandemic GII.4 infected HIE cells show RdRp containing phase separated condensate replication factories. Liquid-to-solid transition triggers the formation of fibrils and inhibits HuNoV pandemic GII.4 RdRp activity. | https://www.biorxiv.org/content/10.1101/2023.08.24.554692v4 |
| TET2 condensates | Leukaemia | Confocal microscopy and FRAP of gfp-tagged TET2 in HEK293T cells show that TET2 undergoes phase separation via its LCI domain. Deletion of the LCI within the endogenous Tet2 locus in mice results in perturbation of endogenous TET2 condensation and confers a suppressive effect on leukaemia cell proliferation. Additionally, PCA of 5hmC signals in mock MOLM-13-cells show TET2 loss of condensation causes chromatin topological changes. | https://pubmed.ncbi.nlm.nih.gov/39251719/ |
| PDE11A condensates | Age-Related Cognitive Decline | COS1 cells transfected with mPDE11A4 fused with EmGFP show mPDE11A4 droplets form in a concentration dependent manner. Treatment with PDE11A4 inhibitors show reduction of LLPS-like clustering of mPDE11A4 in HT22 cells, with clusters reforming following washout of PDE11A inhibitors. Oral dosing of inhibitor SMQ-03-20 in old NIA C57BL6 mice reverse age-related clustering of PDE11A4. | https://www.biorxiv.org/content/10.1101/2024.10.01.616004v1 |
| Stress granules | Amyotrophic lateral sclerosis (ALS) | Upon treatment with sodium arsenite, immunocytochemistry for PABP in iPSC-derived hMNs from ATXN2-patients show stress granule (SG) defects. ATXN2-ALShMNs cells contain larger stress granules during the recovery phase. Additionally, immunohistochemistry for TUJ1 and CHAT on E13 DIV7 primary motor neuron cultures from ATXN2-Q33 TDP-43Tg-ALS mice show altered SG dynamics. | https://pubmed.ncbi.nlm.nih.gov/39209824/ |
| Stress granules | Pancreatic ductal adenocarcinoma (PDAC), Drug Resistance | Immunofluorescence assays of MIA PaC-2 cells show G3BP2 promotes PDAC gemcitabine resistance in a stress granule (SG) dependent manner. TCGA dataset show that G3BP1 and G3BP2 are significantly upregulated in PDAC, with high expression of G3BP2 associated with poor overall survival. mRNA stability assays show SG in G3BP2 knockdown cells under stress have diminished levels of of PDIA3 mRNA. Knockdown of G3BP2 inhibited the proliferation and invasion of PANC‐1 and CFPAC-1 cells in vitro and in vivo. DKC1 was identified as a target of G3BP2, and PDIA3 mRNA stability increased through recruitment to stress granules by G3BP2. | https://pubmed.ncbi.nlm.nih.gov/39289547/ |
| YTHDF2 condensates | Skin Cancer | YTHDF2 protein phase separates in keratinocyte models (HaCaT) during arsenite-induced oxidative stress, promoting the malignant phenotype of keratinocytes. YTHDF2 phase separation is driven by binding of YTHDF2 to multiple m6A sites on PTEN mRNA, recruiting EIF2AK1 and inhibiting PTEN translation initiation and reducing PTEN protein levels. Downregulation of PTEN by YTHDF2 phase separation activates the pro-survival AKT signal. | https://pubmed.ncbi.nlm.nih.gov/39490166/ |
| Stress granules | Amyotrophic lateral sclerosis (ALS) | Acute sublethal sodium arsenite treatment (ARS), as a positive control of oxidative stress, induced TDP-43 mislocalization from the nucleus to the cytoplasm, and cytoplasmic stress granule (SG) formation in human neuroblastoma SK-N-BE cells visualized via confocal microscopy. Semi-quantitative RT-PCR show loss of splicing activity upon chronic ARS treatment in human neuroblastoma cells, which induced features of TDP-43 loss and gain of function associated with TDP-43 proteinopathy. Rapamycin was able to significantly diminish formation of cytoplasmic phosphorylated TDP-43 SG in ALS patient-derived primary fibroblasts. Rapamycin also significantly decreased the percentage of patient-derived mC9orf72 iPSC-MNs with cytoplasmic phosphorylated TDP-43 aggregates from 34.3 % to 23.8 % and the percentage of SGs-forming cells from 12.7% to 4.2%. | https://pubmed.ncbi.nlm.nih.gov/39536963/ |
| Stress granules | Coxsackievirus B3 (CVB3) infection | Immunofluorescence (IF) showed reader proteins YTHDF1 and YTHDF3 colocalize and interact with HuR in stress granules during the early phase of infection. IF showed knockdown of M6A machinery proteins METTL3 or YTHDF2 significantly reduced the number of SGs formed during CVB3 infection. CVB3 3D and dsRNA visualized via IF of RNA-binding protein HuR are stored in SGs and colocalize with HuR at the early infection phase. | https://pubmed.ncbi.nlm.nih.gov/39597541/ |
| Stress granules | Traumatic brain injury (TBI) | Traumatic brain injury (TBI) in Drosophila causes induction of stress granules (SGs). These TBI-induced SGs are persistent, stain positively for TDP-43, p62 and ubiquitin. The degree of SG induction correlates with the level of trauma and the level of mortality. | https://pubmed.ncbi.nlm.nih.gov/29432563/ |
| Stress granules | Amyotrophic lateral sclerosis (ALS); Frontotemporal dementia (FTD) | Patient-derived fibroblasts expressing mutant TDP-43 or C9ORF72 repeat expansion formed more stress granules (SGs) compared to healthy controls upon exposure to sub-lethal levels of oxidative stress. Under chronic oxidative stress conditions, SG form at a lower level in disease versus healthy fibroblasts. TDP-43 co-localizes with SG only under chronic, but not acute stress conditions. Cells under chronic stress form phospho-TDP-43 fibrils and impair the autophagy pathway. | https://pubmed.ncbi.nlm.nih.gov/32827688/ |
| Stress granules | Perioperative neurocognitive disorders (PND) | Co-staining for hnRNPA2/B1 and TIA1 show hnRNPA2/B1 localizes to stress granules (SGs) in aged mild cognitive impairment (MCI) rats after surgery with diverse anaesthetics. Confocal imagery show on the second postoperative day of MCI rats, hnRNPA2/B1 in the hippocampus of rats in the Scramble-shRNA group translocated to the nucleus and form hnRNPA2/B1-SGs in the perinuclear area. FRAP of rat primary hippocampal neurons transfected with RUVBL2-siRNA or NC-siRNA and mCherry-hnRNPA2/B1 shows that RUVBL2 mediates the transition of hnRNPA2/B1 SGs from the hydrogel phase to the liquid phase. Deffective clearance of SGs formed after sevoflurane exposure requires RUVBL2 knockdown. | https://pubmed.ncbi.nlm.nih.gov/39610020/ |
| Stress granules | Dilated cardiomyopathy (DCM) | Immunofluorescence staining of RBM20 mutations in H9c2 cells show that unphosphorylatable amino acid substitutes S638A, S640G, and S638/S640G facilitate export of RBM20 to the cytoplasm and assembly of ribonucleoprotein (RNP) granules. Phosphomimetic mutations S638D and S640D did not restore the nuclear localization. The non-phosphorylation site mutation P641L promoted RBM20 relocalization and the assembly of RNP granules. Transcriptome profiling with deep RNA-seq in RBM20 S637A KI male mice show altered gene expression associated with cardiac dysfunction and RNP granule assembly. | https://pubmed.ncbi.nlm.nih.gov/35394688/ |
| CPEB4 condensates | Idiopathic autism spectrum disorder (ASD) | Fluorescence microscopy imaging of live ex vivo mEGFP–CPEB4 knock-in mouse model primary striatal neurons show a granular distribution of CPEB4 in the cytoplasm. Mouse N2a cells show neuron-specific and SRRM4-dependent regulation of CPEB4 alternative splicing, with neuronal differentiation induced by retinoic acid. FRAP shows that overexpression of full-length GFP-tagged nCPEB4 in N2s cells led to the formation of condensates equivalent to foci formed by endogenous CPEB4 in neurons. Depolarization of ex vivo neurons using KCL or NMDA led to dissolution of the foci. N2a cells were depolarizated by KCL but not NMDA. | https://www.nature.com/articles/s41586-024-08289-w |
| CRTC2 condensates | Autosomal Dominant Polycystic Kidney Disease (ADPKD) | Immunofluorescence dispersed CRTC2 cytoplasmic staining in normal kidneys but clear nuclear staining with prominent punctate structures in the cyst-lining cells in ADPKD mouse kidneys and kidneys from ADPKD patients. CRTC2 and CycT1 display distinct cellular distribution in WT kidneys but colocalize in the nucleus and exhibited discrete puncta staining in ADPKD kidneys. TY1-WT-CRTC2 exhibit nuclear puncta by immunofluorescence staining, whereas TY1-CRTC2-R>A mutant showed diffuse staining, supporting the idea that R>Amutation in CRTC2 disrupts its ability to form condensates in ADPKD cells. ADPKD mouse model show that deletion of Crtc2 caused decreases in kidney size, ratio of kidney weight to body weight, cystic index, and BUN. | https://pubmed.ncbi.nlm.nih.gov/35037420/ |
| MRV condensates | Mammalian reovirus (MRV) | Low-magnification cryoEM images of MRV-infected LLC-MK2 derivative cell lamellae at 6h show multiple small viral condensates in dispersed spots near host cellular machineries, vs 48h where one large condensate appears as a para-chrystalline array of viral particles. Tomograms resolve multiple cellular structures around or within condensates. Analysis of tomograms reveal viral assembly intermediates within the condensate. High-resolution structures of the assembly intermediates were obtained through cryoEM on cellular extracts containing viral particles at 6h and 48h. CryoET of MRV-infected LLC-MK2 derivative cells show that viral factories are membrane-containing cellular condensates introduced by and used for viral replication. CryoEM maps and the atomic model show the functional advantage of conformational changes in MRV-infected cellular condensates in viral replication. | https://pubmed.ncbi.nlm.nih.gov/39639006/ |
| Stress granules | Pulmonary Hypertension | LINC00599 co-localizes with G3BP1 in stress granules, and is elevated in hypoxic PASMCs and pulmonary arteries of SuHx mice. Overexpression of LINC00599 with mutated m6A sites does not alter the enhanced effects observed with LINC00599 shRNA in SuHx mice. LINC00599 facilitates pulmonary hypertension via proliferation of artery smooth muscle cells by hypoxia-induced LLPS through m6A modification. | https://www.biorxiv.org/content/10.1101/2024.12.20.629439v1 |
| Stress granules | African swine fever (ASF) infection | African swine fever virus (ASFV) inhibits stress granule formation in Porcine pulmonary alveolar macrophages (PAMs) despite SG induction via Ars. ASFV increases intracellular GSH by upregulating mitochondrial pAIFM1, and inhibition of ferroptosis pathway factors. GSH inhibitor BSO and activator NAC promotes SG formation in mock-infected PAMs. | https://pubmed.ncbi.nlm.nih.gov/39707514/ |
| Transcriptional condensates | Inflammatory colitis | TFEB formed condensates under nutrition deprivation, and the N terminus CC domain is required for LLPS. Deletion of this domain inhibited TFEB-mediated target gene expression. TFEB condensates co-localize with EP300 and MED1 forming a transcriptional hub. TFEB inhibitors suppress TFEB puncta formation and abolish IF-mediated amelioration of DSS colitis. | TFEB formed condensates under nutrition deprivation, and the N terminus CC domain is required for LLPS. Deletion of this domain inhibited TFEB-mediated target gene expression. TFEB condensates co-localize with EP300 and MED1 forming a transcriptional hub. TFEB inhibitor suppress TFEB puncta formation and abolish IF-mediated amelioration of DSS colitis. |
| Ebola viral factories | Ebola virus infection | Infected Huh7 cells undergo dramatic morphological changes concomitant with nucleocapsid (NC) assembly, and have viral factory (VF) condensates which grow over time, associating with vimentin. Loosley coiled NCs were curved, indicating flexibility, and upon interaction with viral components, condense to thick-walled rigid cylinders in cytoplasmic VFs and a reduction in condensate fluidity. | https://pubmed.ncbi.nlm.nih.gov/39742805/ |
| TRIM8 condensates | Neuro-renal syndrome | TRIM8 truncations and disease variants disrupt condensate formation and lead to abberant protein quality control in immortalized human podocytes. TRIM8 IDR supports condensation in a condensate-defective TRIM1 construct and TRIM8 IDR function can be rescued partially by the alternative FUS IDR. TRIM8 condensation is required for TAK1/NFκB signaling and this signaling is disrupted by TRIM8 disease variants. Patient variant TRIM8 levels were not increased by MG132, and not decreased by cyclohexamide. TRIM8 mutant mice exhibit increased matrix deposition but not changes in glomerular matrix, constent with TRIM8 syndrome phenotype in humans. | https://www.biorxiv.org/content/10.1101/2025.01.02.630836v1 |
| DeaD condensates | Antibiotic-tolerant persistance | Upon serine deprivation the (cognate amino acid for seryl-tRNA synthetase inE. coli KLY cells), SerRSt aberrantly translocates into DeaD condensates via selective recruitment through LLPS. Aberrant SerRSt recruitment impaired localized translation and perturbed translation networks in DeaD condensates, precipitating growth arrest and dormancy in SerSt strains, triggering persistance. | Upon serine deprivation the (cognate amino acid for seryl-tRNA synthetase inE. coli KLY cells), SerRSt aberrantly translocates into DeaD condensates via selective recruitment through LLPS. Aberrant SerRSt recruitment impaired localized translation and perturbed translation networks in DeaD condensates, precipitating growth arrest and dormancy in SerSt strains, triggering persistence. |
| Stress granules | C9orf72-linked Amyotrophic Lateral Sclerosis (C9-ALS); Frontotemporal dementia (FTD) | Arginine-rich dipeptide repeat proteins (R-DPR) binds to N-terminal fragments of FUS to promote phase separation. Kapβ2 prevents and reverses the aggregation and LLPS of wild-type FUS in the presence of arginine-rich dipeptide repeats in vitro. NLS-binding deficient Kapβ2W460A:W730A is more efficient in preventing poly(GR) recruitment into stress granules, reduces (GR)100 aggregation and counteracts neuronal toxicity of poly(GR) in primary cortical neurons without affecting normal FUS phase separation. | https://pubmed.ncbi.nlm.nih.gov/39747573/ |
| Stress granules | Ovarian cancer | "Site-specific MARylation of RACK1 is required for stress granule assembly, and depletion of TARG1 enhances stress granule assembly by increasing RACK1 MARylation. RACK1 mutant inhibits the formation of G3BP1 foci in OVCAR3 cells, and both knockdown and chemical inhibition of PAPR14 reduces RACK1 MARylation and reduces stress granule assembly, showing PARP14-mediated, site-specific MARylation of RACK1 drives stress granule assembly. Loss of RACK1 MARylation sensitizes OVCAR3 cells and OVCAR3 xenograft tumors grown in immunodeficient mice to stress and inhibits their growth. " | https://pubmed.ncbi.nlm.nih.gov/39760726/ |
| Stress granules | Myocardial Ischemia-Reperfusion (MIR) Injury | Sephin1 increased G3BP1 and SGs in cardiomyocytes, but inhibited the expression levels of GADD34 in mice and H9c2 cells with ischemia-reperfusion. Sephin1 prolonged integrated stress response and suppresses global protein synthesis and reduces infarct area in the heart of MIR mice. | Sephin1 increased G3BP1 and SGs in cardiomyocytes, but inhibited the expression levels of GADD34 in mice and H9c2 cells with ischemia-reperfusion. Sephin1 prolonged integrated stress response and suppresses global protein synthesis and reduces infarct area in the heart of MIR mice. |
| P bodies | Multiple sclerosis (MS) | PSMB8 undergoes alternative splicing to i2R-PSMB8 in MS white matter lesions (WMLs). Quantification of EDC4 indicate induced P-body formation in primary human cortical astrocytes, leading to phosphorylation of UPF1 and thus increased nonsense-mediated decay (NMD). P-body load is increased in postmortem MS lesions within astrocytes, potentially indicating a dysregulation in splicing of many genes within the MS white matter lesions. | https://pubmed.ncbi.nlm.nih.gov/38812791/ |
| Stress granules | Kanamycin-induced ototoxicity | METTL3 expression increases kanamycin-induced SG formation and attentuates apoptosis and ROS generation in kanamycin-treated HEI-OC1 cells. Kanamycin treatment in hair cells and HEI-OC1 cells cause decreases in METTL3 expression, and cells isolated from cochlear explants of P3 C57BL/6 mice also show decreased METTL3 expression. Administration of aminoglycosides to mice induce SG formation and decrease expression of METTL3 in the cochlea. | https://pubmed.ncbi.nlm.nih.gov/39193335/ |
| PARP1-DNA condensates | Genotoxic stress | ANGPTL8 interacts with PARP1 in a PARylation-independent manner, reducing fluidity of PARP1-DNA condensates, sensitizing cells to genotoxins in a PARP1-dependent manner. ANGPTL8 also exacerbates the accumulation of PARP1 and PAR chains on chromatin, inhibiting the fluidity of PARP1-related LLPS condensates in HepG2 cells and stabilizes the PARP1-DNA condensates in vitro. Angptl8 deficiency in mice alleviates hepatic I/R-induced acute liver injury. | https://pubmed.ncbi.nlm.nih.gov/39592710/ |
| Stress granules | Retinal degeneration; Retinopathy | SGs were detected in retinal ganglion cells in rat models, and exhibited heightened SG formation in response to prolonged light exposure. | https://pubmed.ncbi.nlm.nih.gov/39813056/ |
| Paraspeckles | Type 2 diabetes (T2D) | Paraspeckle dynamics regulate gluconeogenesis in hepatocytes. Diabetes-relevant proline levels inhibit formation of paraspeckles in hepatocytes via suppression of Neat1 expression by NONO binding, and promotes hepatic gluconeogenesis through paraspeckle mRNA retention axis in mice. Proline selectively regulated paraspeckle mRNA retention of metabolism-related mRNA and expression of PGC1α and FOXO1 hepatocytes, with Ppargc1a and Foxo1 mRNAs colocalizing with Neat1 in the nucleus. Modulating paraspeckle dynamics ameliorates hyperglycaemia in diabetic mice. | https://pubmed.ncbi.nlm.nih.gov/39820557/ |
| Stress granules | Dilated cardiomyopathy (DCM) | Immunofluorescent staining in paraffin-embedded tissues and isolated single cardiomyocyte show an RBM20 S639G mislocalizes from nucleus to the sarcoplasm, with formation of protein condensates in the sarcoplasm. Differential expression and alternative splicing analyses show that components of stress granules were dysregulated in RBM20S639G mice when compared to WT, and genes associated with translational regulation were also differentially expressed in RBM20S639G mice. In vivo cardiac functional assessments show increased heart to body weight ratios in mutant two-month-old homozygous mice, but not in heterozygous mice. | https://pubmed.ncbi.nlm.nih.gov/35041844/ |
| RdRp condensates | Norovirus infection | RdRp of pandemic GII.4 HuNoV forms condensates in infected HIE cells, and is ablated by knockdown of the NTD region in vitro. Uninfected HIEs did not form condensates. Colocalization of dsRNA and VPg indicate condensates as sites for viral replication. Concentration of RdRp is ~20 times higher in condensates than in the dilute phase, and is enzymatically active. | https://pubmed.ncbi.nlm.nih.gov/39705355/ |
| YTHDF2 condensates | Lung cancer | CircDNA2 Promotes YTHDF2 LLPS by binding to the YTHDF2 LC domain in HBE cells. circDNA2 OEX significantly increased YTHDF2 puncta formation, whereas circDNA2 KD inhibited YTHDF2 puncta formation in HBE cells. MS2-tagged pull-down assays show ircDNA2 inhibits GADD45A expression by interacting with the LC Domain of YTHDF2. CircDNA2 Promotes PM2.5 (particulate matter 2.5)-induced malignant transformation by inhibiting GADD45A expression in xenograft murine models. Overall, abnormally upregulated circDNA2 inhibited GADD45A mRNA in an m6A-dependent manner, mediated by YTHDF2 after PM2.5 exposure. | https://pubmed.ncbi.nlm.nih.gov/39823477/ |
| Stress granules | DDX3X syndrome; Autism Spectrum Disorder | DDX3X missense mutations cause formation of stress granules in primary neural progenitors and neurons. The DDX3X missense granules show differential protein composition and physical properties, with C-terminal missense mutations exhibiting reduced molecular exchange with the cytoplasm. Clinically severe DDX3X missense mutations induce DNA damage in neurons and accumulation of R-loops in the cytoplasm. | DDX3X missense mutations cause formation of stress granules in primary neural progenitors and neurons. The DDX3X missense granules show differential protein composition and physical properties, with C-terminal missense mutations exhibiting reduced molecular exchange with the cytoplasm. Clinically severe DDX3X missense mutations induce DNA damage in neurons and accumulation of R-loops in the cytoplasm. |
| HIV-1 membraneless organelles (HIV-1-MLOs) | Human immunodeficiency virus type 1 (HIV-1) infection | HIV-1 membraneless organelles (HIV-1-MLOs) persist in macrophage-like cells (THP-1) in the presence of nevirapine, and build during in vivo infection of mice. Immune RNA-FISH and the MCP-MS2 system show that CPSF6 condensates co-exist with active proviruses and shield viral RNA genome. CPSF6 condensate disassembly impedes nuclear viral reverse transcription after nevirapine removal in THP-1 cells. HIV-1 MLOs shield the final reverse transcription product from host immune detection. | https://pubmed.ncbi.nlm.nih.gov/39623137/ |
| GCRV viroplasm | Grass carp reovirus (GCRV) | LLPS of NS80 drives formation of GCRV viroplasms, and an NS80 mutant and a reagent inhibts LLPS. Viroplasms (VL) form membraneless organelles which coalesce within the cytoplasm of GCRV-infected cells where liquid-like properties recruit viral dsRNA, viral RdRp, and viral proteins. The liquid-like properties of VLs are essential for viral replication, assembly, and immune evasion, and LLPS is essential for VLs to recruit viral proteins for virion assembly. GCO cells transfected with NS38-mCherry plasmid and infected with GCRV showed that most viral dsRNA was concentrated within VLs in the presence of GCRV. In the absence of GCRV infection, EGFP-tagged viral proteins distribute uniformly in the cytoplasm or throughout cells, except for NS80-EGFP which formed VLs. However, in transfected cells infected with GCRV, all EGFP-tagged viral proteins co-aggregate within NS38-mCherry VLs. | https://pubmed.ncbi.nlm.nih.gov/39194247/ |
| IBDV viral factories | Infectious bursal disease virus (IBDV) | The infectious bursal disease virus (IBDV) VP3 C terminal IDR promotes the formation of cytoplasmic puncta and influences their size and shape in DF-1 cells, where puncta formed in the presence of wt VP3 had significantly larger average area and perimeter, greater mean fluorescence intensity, and greater single-puncta intensity than puncta formed in VP3ΔC mutants lacking the intrinsically disordered region (IDR). The VP3 C terminal IDR significantly contributes to the mobile fraction of the molecules within the cytoplasmic puncta, and puncta formed are sensitive to aliphatic diol treatment. | https://www.biorxiv.org/content/10.1101/2025.01.17.633518v1 |
| Aβ42 condensates | Alzheimer's Disease (AD) | Aβ42 undergoes LLPS on the membrane surface of human neuroblastoma SH-SY5Y cells, and matures into aggregates over time. Under conditions where LLPS was observed, aggregates appeared within the condensates earlier than under non-LLPS conditions in the absence of a lipid membrane. Aggregates appeared within the condensates earlier than in non-LLPS conditions, and LLPS opens a more energetically favorable aggregation pathway for Aβ42. | https://pubmed.ncbi.nlm.nih.gov/39854227/ |
| ARAF condensates | RAS-related cancer drug resistance | ARAF, but not isoforms BRAF and CRAF forms punctate structures at the plasma membrane (PM) in SKBR3 (HER2-amplified human breast cancer) cells. PM fractionation showed that endogenous ARAF displayed a higher PM/cytosol ratio than BRAF and CRAF. The disordered N-terminal sequence of ARAF drives self assembly, forming ARAF-RAS condensates tethered to the PM. Condensates concentrate active RAS locally, impeding NF1-mediated negative regulation of RAS, thus fostering receptor tyrosine kinase (RTK)-triggered RAS activation. ARAF N terminus loss boosts effect of RAF dimer inhibitors in A549 and MIA PaCa-2 cell lines. Heregulin-1β (HRG) treatment in MCF7 cells activated HER3 signaling, induced a gradual increase in membrane condensation of ARAF, and caused downregulation of ER expression, which attenuated the growth inhibition in cells treated with fulvestrant. ARAF knockdown curbed cell proliferation and re-sensitized the cells to fulvestrant treatment. | https://pubmed.ncbi.nlm.nih.gov/39870764/ |
| Nucleolus | Lung adenocarcinoma | USP39 is a deubiquitylating enzyme that undergoes LLPS in the nucleolus through the N-terminal disordered region. USP39 silencing leads to growth and migration inhibition in lung adenocarcinoma A549 and H1299 cell lines. MTT, colony formation and transwell assays show that absence of the USP39 IDR region and the C-terminal domain damages the oncogenic activities of USP39 in lung adenocarcinoma. Transcriptomic analysis revealed that phase-separated USP39 was associated with GLI1 expression. Western blots of lung cancer and adjacent normal tissue pairs show elevated protein levels of USP39 and GLI1. Analysis of the association between patient survival and mRNA levels show high expression of GLI1 were associated with poorer prognosis. USP39 knockdown suppresses GLI1 expression, and USP39 knockdown effectively hinders xenograft growth in athymic nude mice. | USP39 is a deubiquitylating enzyme that undergoes LLPS in the nucleolus through the N-terminal disordered region. USP39 silencing leads to growth and migration inhibition in lung adenocarcinoma A549 and H1299 cell lines. MTT, colony formation and transwell assays show that absence of the USP39 IDR region and the C-terminal domain damages the oncogenic activities of USP39 in lung adenocarcinoma. Transcriptomic analysis revealed that phase-separated USP39 was associated with GLI1 expression. Western blots of lung cancer and adjacent normal tissue pairs show elevated protein levels of USP39 and GLI1. Analysis of the association between patient survival and mRNA levels show high expression of GLI1 were associated with poorer prognosis. USP39 knockdown suppresses GLI1 expression, and USP39 knockdown effectively hinders xenograft growth in athymic nude mice. |
| Lactate bodies | Hepatic fibrosis | SORBS3 lactylation promotes phase separation in C2C12 myocytes. The K479R SORBS3 mutant did not. Upon 1,6-hexanediol treatment, there was a diminished FBXO2-FLOT1 interaction in C2C12 cells and less FBXO2 content in small extracellular vesicles (SEVs). LLPS of SORBS3 enhanced its interaction with flotillin 1 and selectively facilitates the sorting of FBXO2 into SEVs, referred to as "lactate bodies." During overtraining in mice, Sorbs3 KO reduced the proportion of lactate bodies in the culture suspension of muscle tissues. Overtrained Sorbs3-MKO mice exhibited an alleviation of liver injury, collagen accumulation, and expression of fibrosis-related genes compared with overtrained WT mice. Lactate bodies induce liver fibrosis via an MCL1-BAX/BAK-dependent pathway, and treatment with salidroside inhibited LDH activity and reduced lactate levels in muscle tissues of mice. | https://pubmed.ncbi.nlm.nih.gov/39879982/ |
| EEF1E1 condensates | Hepatocellular carcinoma | EEF1E1-GFP forms condensates in HCCLM3 cells which are dissolved by 1,6 hexanadiol 1,6-HD). EEF1E1 silencing via 1,6-HD treatment led to reduced expression of stem cell markers CD133, EpCAM, and SOX2, quantified via Western blot analysis. Overexpression of EEF1E1 caused inactivation of PTEN and activation of p-AKT which were significantly reversed after adding 1,6-HD. EEF1E1 silencing in xenograft tumor mice models showed weakened tumor growh, reduction in expression of tumor cell proliferation marker Ki67, decreased expression of CD133, EpCAM, and SOX2, and activation of PTEN and reduction of p-AKT. | https://pubmed.ncbi.nlm.nih.gov/39884379/ |
| Stress granules | Alzheimer's Disease (AD) | Aβ42 causes SG formation in a cell type-specific manner and depends on eIF2α phosphorylation. Exposure to 20 μM Aβ42 led to significant SG formation in neuroblastoma-derived (SH-SY5Y) and glioma-derived (U87) cell lines, but not non-neuronal cell lines. Also, familial mutants Dutch E22Q and Flemish A21G display distinct SG-inducing characteristics. Monomeric and oligomeric forms of Aβ42 exhibit a 4–5 times stronger ability to induce SG formation compared to fibrillar forms. eIF2α kinases knockout (KO) cell lines show that Aβ-induced SG formation is dependent on Protein Kinase R (PKR). Proximity ligation assay (PLA) analysis show a close proximity of PACT and PKR in both Aβ-treated SH-SY5Y and U87 cells, and in Alzheimer's Disease (AD) mouse hippocampus indicating Aβ42 exposure facilitates PACT-PKR interaction. | Aβ42 causes SG formation in a cell type-specific manner and depends on eIF2α phosphorylation. Exposure to 20 μM Aβ42 led to significant SG formation in neuroblastoma-derived (SH-SY5Y) and glioma-derived (U87) cell lines, but not non-neuronal cell lines. Also, familial mutants Dutch E22Q and Flemish A21G display distinct SG-inducing characteristics. Monomeric and oligomeric forms of Aβ42 exhibit a 4–5 times stronger ability to induce SG formation compared to fibrillar forms. eIF2α kinases knockout (KO) cell lines show that Aβ-induced SG formation is dependent on Protein Kinase R (PKR). Proximity ligation assay (PLA) analysis show a close proximity of PACT and PKR in both Aβ-treated SH-SY5Y and U87 cells, and in Alzheimer's Disease (AD) mouse hippocampus indicating Aβ42 exposure facilitates PACT-PKR interaction. |
| Stress granules | Nerve injury | Corticospinal axons in injured spinal cord contain more G3BP1-TIA1 granules in mice with a rostralized progenitor graft, which does not support regeneration, than those with a caudalized progenitor graft. Immunofluorescence of axons performed at 20 days after spinal cord injury/peripheral nerve graft (PNG) placement showed more axons extending further distances from the spinal cord/PNG apposition in the G3BP1 B-domain-BFP expressing mice compared to the G3BP1-BFP and GFP expressing mice. Additionally, immunofluorescence of retinal ganglion cells (RGCs) from an optic nerve injury mouse model showed that G3BP1 B-domain expression in RGCs greatly increases axon regeneration. G3BP1 B-domain expression in reticulospinal neurons and RGCs resulted in smaller and fewer G3BP1 granules in the axons compared to G3BP1 and control GFP expression, and transduction with AAV2-G3BP1 B-domain-BFP increased postinjury RGC survival compared to controls. Also, injection of a cell-permeable G3BP1 B-domain peptide increases nerve regeneration in mouse mid-thigh sciatic nerve crush injuries. G3BP1 192-205 (EP7) CPP significantly reduces both the density and size of axonal G3BP1 granules in embryonic rat cortical neurons. | https://pubmed.ncbi.nlm.nih.gov/40014573/ |
| PABPN1 condensates | Colon cancer | SW480 cell lines stably expressing Cas9 and PABPN1-BiFC shows higher percentage of PABPN1-BiFC within condensates in cells with SNRPD2 knockdown than in control cells, confirming that SNRPD2 can repress the LLPS of PABPN1. Colorectal cancer tumor suppressor QKI-6 promotes PABPN1 LLPS in SW480 and SW620 cells, with QKI-6-G118R and QKI-6-ΔKH mutants losing the ability to promote LLPS. CCK-8 and Transwell assays show increased cell proliferation and migration rates in knockdown of QKI in SW480 cells. Overexpression of QKI-6 in SW480 and SW620 significantly inhibits both cell proliferation and migration. SW480 and SW620 cells co-transfected with QKI-6 and PABPN1 WT or LLPS-deficient mutant show inhibitory effects of QKI-6 on proliferation and migration in the LLPS-deficient mutant compared to WT. Additionally, IF imaging analysis reveals QKI-6-nuclear localization signal exhibit a significantly higher percentage of PABPN1 in LLPS droplets compared to those overexpressing QKI-6-WT. | https://pubmed.ncbi.nlm.nih.gov/40052530/ |
| Nuclear speckles | Breast cancer | ZNF32 H179A, H183A, and H179A, H183A mutations, but not WT, form nuclear speckles (NS) in ZR-75-30, MCF-7, and MDA-MB-231 breast cancer cells. MTT assay, chrystal violet staining, and EdU staining show ZNF32 single-site mutations (H179A, H183A) but not ZNF32 H179A, H183A and WT promote the proliferation of cells. Subcutaneous xenograft tumor model in nude mice show higher tumor volumes and tumor formation rates in H179A and H183A single-site mutations, but not ZNF32 H179A, H183A double mutations or WT group. Transcriptome sequencing data show ZNF32 H179A and H183A differentially upregulate ISY1-RAB43 and UPK3BL1 as well as downregulating SNX22 expressions. | https://pubmed.ncbi.nlm.nih.gov/40046230/ |
| Stress granules | Hepatocellular carcinoma | Missense mutp53 conferred better overall survival to hepatocellular carcinoma (HCC) patients compared to p53null patients. Knockdown of mutp53 in Huh7 (p53Y220C), SNU449 (p53A161T), and HepG2 (p53wt) HCC cells or overexpressed p53R175H-3XFLAG in p53null Hep3B HCC cells treated with sorafenib (SOR), vincristine (VCR), doxorubicin (DRB), or cisplatin (CDDP) reveal mutp53-expressing Huh7, SNU449, and Hep3B cells showed increased apoptosis compared to p53-knockdown or -null cells following SOR and VCR treatment, but not DRB or CDDP treatment. Additionally, immunofluorescence for CC3 and colony formation assays show mutp53-expressing KHOS/NP cells had more apoptotic cells with reduced colony formation than mutp53-knockdown cells treated with ER-stress inducer, thapsigargin (TG). Overexpression of p53R175H in p53null Hep3B HCC cells and OSC19 head and neck cancer cells resulted in increased apoptosis following TG treatment. Knockdown or knockout of mutp53 significantly increased the number of SG-positive cells following TG treatment in KHOS/NP, PLC/PRF/5 (p53R249S, HCC), and SNU449 cells. Knockdown of mutp53 in SNU449 and Huh7 HCC cells resulted in a consistent and significant increase in eIF2α phosphorylation (p-eIF2α) and increase in p-PERK following TG treatment. Increased PABP- or eIF4E SG formation due to mutp53 knockdown was reduced by simultaneous G3BP1 knockdown in both SNU449 and KHOS/NP cells following SOR and TG treatment. Hep3B cells expressing FL p53R175H and deletion mutants treated with SOR reveal that the OD domain in mutp53 is required for the mutp53-G3BP1 interaction. Immunocompromised mice bearing control and mutp53-knockdown Huh7 tumors show that mutp53-expressing control tumors responded to SOR, while mutp53-knockdown tumors show reduced sensitivity to SOR. Immunohistochemistry for p53 and CC3, as well as immunofluorescence for PABP and G3BP1 show increased CC3 positivity and a reduced frequency of SG formation in SOR-treated control tumors compared to mutp53-knockdown tumors. | https://pubmed.ncbi.nlm.nih.gov/40064891/ |
| PML bodies | Ocular melanoma | Anti-m1A dot-blot assay show ocular melanoma samples present attenuated m1A levels and increased ALKBH3 protein levels. Kaplan–Meier curves of tumor recurrence reveal poor outcomes with elevated ALKBH3 levels. ALKBH3-deficient ocular melanoma cells (92.1, OMM2.3 and CRMM1) show an increase of m1A levels, and and attenuation of cell growth, colony forming capacity, and decreased migration capacity. Injection of luciferase-labeled control and ALKBH3-silenced 92.1 melanoma cells into nude mice show a weaker signal intensity in ALKBH3-deficient ocular melanoma cells than in control cells. Correlation analysis of the relative protein expression of ALKBH3 and H3K18la level in ocular melanoma cells and normal melanocytes, and CUT&Tag and ChIP-seq of H3K18la analysis reveal histone lactylation enhances ALKBH3 expression. Western blot, RNA-seq, and qPCR reveal decreased RNA expression level of SP100A in ocular melanoma cells, and immunofluorescence of clinical samples show that SP100A was significantly decreased in the ocular melanoma samples. Overexpression of SP100A result in increased PML protein levels and an increase of PML bodies in ocular melanoma cells. Also, ALKBH3-deficient ocular melanoma cells exhibit an elevation in PML bodies. | https://pubmed.ncbi.nlm.nih.gov/38118002/ |
| Stress granules | Cancer | Immunoflourescence microscopy of U2OS cells show that cells expressing the p53 DBD-depleted mutants have reduced percentages of cells harboring puncta as compared to those expressing p53 WT. Immunoblot analysis of H1299 cells expressing WT or site-specifically lactylated p53 show that the site-specifically lactylated p53 proteins failed to bind p53RE-DNA or condense into discrete nuclear puncta in response to stimuli. To examine the consequences of p53 lactylation, p53-/- HCT116 cells expressing either the empty vector (p53null), wild-type (p53WT), or the K120&K139-lactylated p53 (p53DualLac) were inoculated in mice and monitored for tumor growth. p53Dual-Lac was more tumorigenic than the p53WT, with levels of p53 K120 and K139 lactylation correlated with tumor volume. Additionally, p53-WT, but not the K120E and K139E disease variants, inhibit cellular proliferation, clone formation, and xenograft tumor growth in HCT116 cells. | p53 |
| SRSF9 condensates | Oral cancer | HEK293 cells transfected with mutant SRSF9 delta-arginine-serine-rich (RS) domain show reduced puncta area as compared to WT SRSF9 and truncated variants (SRSF9-△RRM1, SRSF9-△RRM2). SLC37A4 exon 7 minigene reporter system reveals that overexpression of SRSF9-WT, SRSF9-△RRM1, and SRSF9-△RRM2 in knockdown SRSF9 cells significantly rescued knockdown SRSF9-induced SLC37A4 exon 7 inclusion, whereas overexpression of SRSF9-△RS showed no rescue effects. Additionally, SRSF9-WT, SRSF9-△RRM1, and SRSF9-△RRM2 successfully reversed the inhibition of oral cancer cell proliferation and migration due to SRSF9 knockdown, but SRSF9-△RS did not. | https://pubmed.ncbi.nlm.nih.gov/40064440/ |
| FOXP1 condensates | Small cell lung cancer (SCLC) | Immunofluorescence of patient-derived xenograft (PDX) chemotherapy-sensitive or resistant samples reveal that FOXP1 undergoes LLPS in resistant samples. In H69AR cells, PDX2-resistant cells, and primary cells from chemoresistant PDX tissues, endogenous FOXP1’s punctate structures colocalized with endogenous BRD4, and histone markers H3K27Ac and H3K4me1 within the nucleus. Proximity ligation assay confirms FOXP1 combines with other transcription proteins. Treatment with BRD4 inhibitors showed global and concentration dependent inhibition of transcription levels, decreased H3K27Ac and H3K4me1, and reduced protein fluidity detected via FRAP, confirming disrusption of condensates. A patient-derived xenograft model with a control group, single-drug ARV-771 group, single-drug Talazoparib group, and double-drug combination group shows that the combination treatment exhibited the strongest tumor inhibition effect. Immunohistochemical analysis of subcutaneous tumors confirms that FOXP1 protein levels remained unaffected by BRD4 inhibitors, whereas SP8 expression was inhibited. | https://pubmed.ncbi.nlm.nih.gov/40082538/ |
| Perineuronal nets (PNNs) | Neuropathic pain | The density of PNNs labelled with the lectin Wisteria Floribunda Agglutinin (WFA) increases in the contralateral somatosensory cortex (SSC), medial prefrontal cortex (mPFC), reticular thalamic nucleus (RTN), and insular cortex of mice in response to unilateral chronic constriction injury of the sciatic nerve. Immunoblot assay shows that in the SSC and mPFC, increased WFA PNNs are associated with an upregulation of brevican and neurocan. Enzymatic degradation of PNNs via chondroitinase ABC in the contralateral SSC or RTN enhanced both mechanical and thermal pain thresholds in chronic constriction injury mice. siRNA-induced knock-down of the PNN-degrading enzyme MMP-9 in the SSC or RTN lowered pain thresholds in sham-operated mice. | https://pubmed.ncbi.nlm.nih.gov/40112162/ |
| Stress granules | Head and neck cancer (HNC) | Western blotting analysis and immunofluorescence staining of tumor and adjacent non-tumor head and neck cancer (HNC) tissue samples show that protein levels of stress granule (SG) markers G3BP1 and PABP were higher in tumor (T) samples than non-tumor (N) samples, with distinct puncta in the cytosol of T samples only. Immunofluorescence microscopy of FaDu cells show that HSP90 inhibitor 17AGG decreased TBK1 activity and the number of cells displaying SGs in cells experiencing proteotoxic stress induced by MG132, but not in the absence of stress stimuli. Additionally, treatment with chemotherapeutic drug cisplatin significantly induced SG formation, while TBK1 knockdown reduced SG-positive cells by approximately 46%. Additionally, FaDu cells transfected with NC, TBK1, ATG5, or G3BP1 siRNAs treated with cisplatin showed cleaved CASP3 levels were higher in siTBK1- and siG3BP1-treated cells compared to siATG5-treated cells. Use of novel TBK1 inhibitor, GSK8612 suppresses xenograft tumor growth in vivo. p-TBK1 expression in tumors was reduced in GSK8612-treated mice compared to control mice, and immunofluorescence of tumor sections revealed that SGs were significantly reduced in GSK8612-treated tumors compared to control tumors. | https://pubmed.ncbi.nlm.nih.gov/40114316/ |
| LARP7 condensates | Human immunodeficiency virus type 1 (HIV-1) | In Jurkat cells, HIV-1 infection, but not Herpes simplex virus nor Vesicular stomatitis virus induced the formation of LARP7 condensates. Immunofluorescence show HEXIM1 and Cyclin T1 co-concentrate in LARP7 puncta after HIV-1 infection in Jurkat cells. Additionally, Tat foci were observed to co-localize with LARP7 upon HIV-1 infection. In a dual luciferase reporter assay with Tat and a Renilla luciferase reporter plasmid in Tzm-bl cells, transient knockdown of LARP7 led to increased luciferase activity, indicating that LARP7 represses Tat-mediated transcription. This was rescued by expression of wild-type LARP7, but not by the LLPS-deficient 49KA and 30KA mutants. | https://pubmed.ncbi.nlm.nih.gov/40113991/ |
| Transcriptional condensates | Asthma | Immunohistochemical staining reveal ovalbumin (OVA)-induced asthma mice models show colocalization of SUMO1 and TFEB. Immunoprecipitation (IP) of endogenous TFEB SUMOylation in BEAS-2B cells treated with OVA show increased levels of SUMO1 and TFEB. TFEB in AEC cells show increased levels of SUMOylated TFEB after OVA stimulation, suggesting SUMOylation of TFEB is related to allergic airway inflammation. In BEAS-2B cells, TFEB-K361R-EGFP (SUMOylation site mutant) formed increased number of puncta than TFEB-WT-EGFP specify cell line. TFEB-K361R-EGFP also forms increased number of puncta in nuclei after starvation or OVA-stimulation for 6 h, while TFEB-WT-EGFP shows lowered number of puncta formation. Also, nuclear TFEB-K361R forms distinct puncta that colocalize with LAMP1 mRNAs of target genes during the formation of TFEB condensates, suggesting TFEB puncta are involved in transcription. | https://pubmed.ncbi.nlm.nih.gov/39998815/ |
| HNRNPH1 condensates | Colorectal cancer (CRC) | Protein and mRNA expression levels of PTK6 were significantly higher in colorectal cancer human (CRC) tissues compared to normal tissues. Western blot analysis, mCherry-EGFP-LC3 dual fluorescence reporter assay, and flow cytometry reveals that PTK6 activates autophagy and inhibits apoptosis through HNRNPH1. Confocal microscopy images of HCT116 cells transfected with different HNRNPH1-expressing plasmids show that PTK6 regulates the LLPS capability of HNRNPH1 by phosphorylating Y210. HNRNPH1-RNA-seq analysis further show that PTK6 promotes HNRNPH1 LLPS, which leads to NBR1 exon retention, resulting in activation of autophagy and inhibition of apoptosis. LLPS of HNRNPH1 was significantly inhibited in HCT116 and DLD1 cells following tilfrinib treatment. mCherry-EGFP-LC3 dual fluorescence assays demonstrated that tilfrinib significantly inhibits PTK6-mediated promotion of autophagy. PTK6 significantly promotes patient-derived organoid formation, whereas tilfrinib substantially inhibits PTK6-induced tumor proliferation. Additionally, LLPS of HNRNPH1 was significantly inhibited in HCT116 and DLD1 cells following tilfrinib treatment. | https://pubmed.ncbi.nlm.nih.gov/40103198/ |
| PABPC1 condensates | Chronic myeloid leukaemia (CML) | PABPC1 deletion suppresses chronic myeloid leukemia (CML) blast crisis (BC) disease progression in mouse models. Immunofluorescence of primary human cells reveal PABPC1 colocalizes with eIF4G and forms granules throughout the cytoplasm, with more condensates forming in patients with CML-BC. Luciferase reporter assays show that PABPC1 preferentially promoted the translation of luciferase mRNAs with long and highly structured 5′UTRs instead of short and not structured 5′UTRs, and that deletion of IDR3 reduces this promotion, supporting a functional requirement of PABPC1 phase separation to boost target mRNA translation and CML-BC progression. Small molecules 1,10-Phen and ML324 bind to PABPC1 and inhibit its RNA binding activity and prevent condensate formation with poly(A) RNA in vitro, and suppress proliferation in K562 and MEG-01 cells. Additionally, a patient-derived xenograft (PDX) mouse model shows that 1,10-Phen or ML324 treatment prevents disease progression and leads to a decreased percentage of various leukaemia stem progenitor cells in bone marrow, with combination therapies being most effective. | https://pubmed.ncbi.nlm.nih.gov/40102686/ |
| Stress granules | Pancreatic cancer | In human pancreatic cancer cell lines MiaPaCa-2, HPAC, Panc-1, and SU.86.86, individual cancer cells within cell populations show a high degree of heterogeneity in SG levels under various stress stimuli. Single cell analysis across cell-cycle phases in MiaPaCa-2 cells using a Fluorescent Ubiquitination-based Cell-cycle Indicator (FUCCI) system reveal that compared to cells marked as G1- and S113 phase, cells marked as G2-phase had ~6x higher levels of SGs, and S-phase showed a ~1.5x increase in SG levels compared to G1-phase cells. Also, SG levels were higher in G2-phase cells, G1 and S-phase cells showing low levels of SGs. Levels of 15d-PGJ2 were ~10-20x higher in G2-phase cells compared to asynchronous cells, and supplementation of asynchronous cells with increasing concentrations of exogenous 15d-PGJ2 enhanced SG levels up to the levels of G2-phase cells (but supplementation of G2-phase cells with the same concentrations of exogenous 15d-PGJ2 had no impact on SG formation capacity). Treatment of MiaPaCa-2 and HPAC cells with L-PGDS specific inhibitor AT-56 reduced SG formation in ~9x in G2-phase and ~3x in asynchronous cells. Additionally, cPLA2 is specifically required for the enhanced SG formation in G2- 266 phase, and knockdown of cPLA2 by doxycycline-inducible shRNA impaired SG formation in pancreatic cancer cells synchronized in G2-phase, but not in asynchronous cells. In pancreatic cancer cell tissue samples from orthotopic mouse models of pancreatic ductal adenocarcinoma, SG levels were ~3x higher in Geminin positive (S/G2) cells than in Geminin negative (G1) cells indicating that SG formation is influenced by cell-cycle state in vivo. KPC-luciferase cells orthotopically implanted in mice treated with oxaliplatin, cPLA2 inhibitor PYR, or both show an increased SG index in tumors that received oxaliplatin treatment alone, correlating with a higher percentage of G2 phase cells and their ability to promote SG formation. PYR inhibited SG formation in Geminin-positive (S/G2) cells, but not Geminin-negative cells. | https://pubmed.ncbi.nlm.nih.gov/40099649/ |
| FOXM1+ASPM condensates | Hepatocellular carcinoma (HCC) | Genome-wide screen is used to identify intranuclear proteins that promote FOXM1 transcription activity via liquid–liquid phase separation (LLPS). Endogenous FOXM1 and ASPM form condensates and colocalize in HepG2 cells. Knockdown of either FOXM1 or ASPM disrupt condensate formation in HepG2 cells, and knockdown of ASPM reduced the protein level of FOXM1 in SNU-739 and HepG2 cells. Treatment with MG132 thwarted the effect of ASPM silencing on regulation of FOXM1 at the protein level (visualized via Western blot), suggesting ASPM promotes accumulation of FOXM1 by inhibiting its proteasome-mediated degradation. CCK-8 assays show cell viability was reduced with ASPM silencing in SNU-73 cells, but reversed by re-constitutive expression of FOXM1. Overexpression of ASPM in MHCC-LM3 cells enhanced proliferation and clone formation. Additionally, subcutaneous injection of SNU-739 in xenograft mouse models show ASPM-deficient tumors had lower levels in tumor volume, tumor weight, and tumor growth. | https://pubmed.ncbi.nlm.nih.gov/40122889/ |
| TDP-43 inclusions | Amyotrophic lateral sclerosis (ALS) | Sonicated fibril fragments (seeds) were introduced by liposome-based transfection into U2OS cells that expresses fluorescently tagged TDP-43 with a disrupted nuclear localization signal (TDP-43mNLS-Clover) in a doxycycline-inducible manner. In normal conditions, TDP-43mNLS-Clover forms reversible condensates. 1 day after seed exposure, fibril-treated cells accumulated TDP-43mNLS-Clover inclusions positive for the phosphorylated form of TDP-43 at serine 409/410 (a key feature of pathological inclusions found in ALS/FTD patients). Quantification of cells with phosphorylated TDP-43 inclusions and size of inclusions reveal a dose-dependent effect with up to 31% ± 3.7% of the cells accumulating phosphorylated TDP-43mNLS-Clover aggregates. Time-lapse fluorescence microscopy show seeds induce TDP-43 inclusions with distinct morphologies and biophysical properties. | https://pubmed.ncbi.nlm.nih.gov/40157356/ |
| Stress granules | Pseudorabies virus (PRV) | Pseudorabies virus (PRV) infection inhibits SG formation in PK15 and H1299 cells. PK-15 cells infected with PRV show PRV infection promotes G3BP1 nuclear translocation in infected host cells. Confocal microscopy and statistical analyses of SG formation showed that expression of IE180 significantly inhibited poly(I:C)- or arsenite-induced SG formation as compared with US1. Co-immunoprecipitation (co-IP) assays report the interaction between PRV IE180 and endogenous G3BP1 in PRV-infected PK-15 cells, and immunofluorescence assays showed that both GFP-G3BP1 and GFP-G3BP2 colocalized with IE180 protein in the nucleus. H1299 G3BP knockout cell lines show that viral gB protein expression levels were substantially increased in PRV-infected H1299 G3BP1−/− cells compared to WT, and the viral titer in PRV-infected H1299 G3BP1−/− cells was significantly higher than WT. | https://pubmed.ncbi.nlm.nih.gov/40145738/ |
| PRC2 bodies | Triple-negative breast cancer (TNBC) | High-resolution microscopy shows that endogenous PRC2 core subunits accumulate in micron-sized nuclear bodies in bone entrained MDA-MB-231 cells (BoM-1833 sub-population), but not EZH2 and SUZ12 cells. PHF19 is identifed through spatial proteomics as a required component of the PRC2 bodies. Immunostaining of endogenous PHF19 show that 40% of BoM-1833 sub-population cells display an accumulation of PHF19 in distinct nuclear bodies whereas MDA-MB-231 show diffuse localization in the nuclei. Transient PHF19 knockdown demonstrate that PHF19 is critical for maintaining the spatial organization of PRC2 bodies in BoM-1833 cells. Western blot analysis reveals PHF19 knockdown over four days had no significant effect on total H3K27me3 levels, but EZH2 depletion over the same period caused a pronounced reduction in H3K27me3 levels in BoM-1833 and MDA-MB-436 cells. Analysis of a triple-negative breast cancer (TNBC) dataset show elevated PHF19 expression in most tumor samples, with a statistically significant upregulation in TNBC patients. | https://www.biorxiv.org/content/10.1101/2025.03.13.642950v1 |
| P bodies | Glioblastoma multiforme (GBM) | RNA sequencing analysis in U87MG human donor cells reveals that praja2 plays a role in the regulation of the transcriptional landscape in Glioblastoma multiforme (GBM). Proteomic analysis identified the RNA helicase DDX6 as a novel target of praja2 and in situ immunostaining analyis of U87MG cell lines confirm that a portion of praja2 signal partially colocalizes with DDX6 at P-bodies. Co-immunoprecipitation show praja2 co-purified with other components of P-bodies involved in translational repression, mRNA decapping, and RNA degradation. In U87MG and patient derived GBM cells, activation of cAMP stimulated the assembly of P-bodies in wild-type cells, whereas the number of P-bodies was reduced in praja2 KO cells, demonstrating that activation of GPCR-cAMP-PKA-praja2 axis promotes the assembly of P-bodies. Wild-type or praja2KO GBM cells injected into CD1 nude mice shows that tumor volume was markedly reduced in mice injected with praja2 KO cells compared to controls. Immunohistochemistry confirmed a marked reduction of the proliferative marker Ki67 in praja2 KO tumor samples compared to wild-type. Additionally, overexpression of DDX6 all K/R mutant markedly impaired condensation of P-bodies in Fsk-treated U87MG cells, and expression of the DDX6 ubiquitylation mutant strongly accelerated tumor growth in mice compared to wild-type DDX6. | https://pubmed.ncbi.nlm.nih.gov/40148504/ |
| Stress granules | Amyotrophic Lateral Sclerosis (ALS) | In hFUS transgenic mice, the spinal cord at the end stage of the disease (39–41 days) is marked by a significant increase in the expression of hnRNP A2/B1 isoforms lacking exon 9 (A2b and B1b), downregulation of exon 9-containing isoforms (A2 and B1), and skipping of hnRNP A2/B1 exon 9 (31–36 days) occurs concomitantly with the appearance of disease symptoms. Immunofluorescence analysis on mouse primary cortical neurons shows Exon 9 skipping causes redistribution of hnRNP A2/B1 from the nucleus to the cytoplasm, and Exon 9 skipping increases hnRNP A2/B1 recruitment to SGs. | https://pubmed.ncbi.nlm.nih.gov/40157939/ |
| PSD95 condensates | Fragile X Syndrome (FXS) | In CHO cells, docosahexaenoic acid (DHA) leads to increased PSD95 protein scaffold condensation independent of canonical fatty acid receptor 4 signaling, facilitating re-localization of cytosolic “full-droplet” condensates composed of the key synaptic elements PSD95 and Kv1.2 to the plasma membrane as “half-droplets.” Juvenile wild-type and Fmr1 KO mice, modeling human fragile X syndrome (FXS), under DHA-enriched or -depleted diets show that DHA reverses the inhibitory overtone by promoting the re-localization of presynaptic PSD95-Kv1.2 condensates to interneuron terminal membranes and corrects morpho-functional synaptic defects and stereotypic behaviors. | https://pubmed.ncbi.nlm.nih.gov/40232934/ |
| Stress granules | Dilated cardiomyopathy (DCM) | Genetic variants in the nuclear localization signal of RBM20 hinder its nuclear import and formation of promote cytoplasmic pathogenic RNP granules. Reducing the level of RBM20 expression through treatment with antisense oligonucleotides (ASOs) significantly decreased cytoplasmic RNP granules within RBM20S639G cardiomyocytes. In RBM20S639G (serine-to-glycine substitution in the nuclear localization signal of RBM20) mice, ASO treatment reversed cardiac dysfunction and remodeling as shown by improving ejection fraction, decreasing severity of left ventricular chamber dilation, mitigating left ventricular hypertrophic remodeling, and improving ECG parameters. | https://pubmed.ncbi.nlm.nih.gov/40242865/ |
| PrPC condensates | Chronic kidney disease (CKD) | PrPC condensates are required to activate TBK1-IRF3 signaling in NRK-52E cells. In PTECs, PrPC promoted a profibrotic response via activation of TANK binding kinase 1 (TBK1)–interferon regulatory factor 3 (IRF3) signaling via LLPS. Functional enrichment analysis using DAVID (the Database for Annotation, Visualization, and Integrated Discovery) show that PrPC provokes a profibrotic response in renal tubular cells via the TBK1-IRF3 pathway. Immunohistochemical staining of human renal biopsies show PrPC was up-regulated in fibrotic renal lesions, with renal expression of PrPC positively correlated with the severity of renal failure and the decline in estimated glomerular filtration rate from patients with chronic kidney disease (CKD). In mice, tubular-specific deletion of PrPC mitigated renal fibrosis induced by unilateral ureteral obstruction (UUO) or unilateral ischemia-reperfusion injury (UIRI). Treating mice with TBK1 inhibitor amlexanox mitigated worsening of renal fibrosis and renal function. | https://pubmed.ncbi.nlm.nih.gov/40238918/ |
| TFE3-NONO condensates | Translocation renal cell carcinoma (tRCC) | TFE3 fusion oncoproteins (FOs) form biomolecular condensates in U-2 OS cells with the coiled-coil domains (CCDs) of NONO and SFPQ essential for condensate formation. Comparing GPNMB mRNA levels in U-2 OS cells expressing wild-type TFE3, TFE3 FOs, and various domain deletion and CCD mutants of TFE3 FOs show TFE3 FOs bind to new chromatin regions, alter chromatin accessibility, and form new enhancers and super-enhancers at pro-growth gene loci, and disruption of condensate formation via CCD modification abolishes genome-wide changes. Prominent nuclear puncta/condensates also formed in mouse xenograft tumor models and in patient-derived translocation renal cell carcinoma (tRCC) cell lines, with TFE3 FO condensates mediating cell proliferation, migration, and oncogenic trancriptional programs in tRCC cell lines. | https://pubmed.ncbi.nlm.nih.gov/40222010/ |
| TFE3-SFPQ condensates | Translocation renal cell carcinoma (tRCC) | TFE3 fusion oncoproteins (FOs) form biomolecular condensates in U-2 OS cells with the coiled-coil domains (CCDs) of NONO and SFPQ essential for condensate formation. Comparing GPNMB mRNA levels in U-2 OS cells expressing wild-type TFE3, TFE3 FOs, and various domain deletion and CCD mutants of TFE3 FOs show TFE3 FOs bind to new chromatin regions, alter chromatin accessibility, and form new enhancers and super-enhancers at pro-growth gene loci, and disruption of condensate formation via CCD modification abolishes genome-wide changes. Prominent nuclear puncta/condensates also formed in mouse xenograft tumor models and in patient-derived translocation renal cell carcinoma (tRCC) cell lines, with TFE3 FO condensates mediating cell proliferation, migration, and oncogenic trancriptional programs in tRCC cell lines. | https://pubmed.ncbi.nlm.nih.gov/40222010/ |
| Stress granules | Metabolic Dysfunction-Associated Fatty Liver Disease (MAFLD) | A gene knock-down model of G3BP1, a core SG molecule, in mice and HepG2 cells was constructed to explore the role of SGs in metabolic dysfunction-associated fatty liver disease (MAFLD) induced in vivo by a high-fat diet or in cellulo by palmitic acid (PA). G3BP1 and TIA1 expression were upregulated in both PA-induced HepG2 cells and high-fat diet-fed mouse liver tissues, and showed increased colocalization in the livers of obese but not lean mice. G3BP1 deficiency aggravated liver lipid deposition and insulin resistance in obese mice, confirmed via PA-induced hepatocytes. RNA-seq show G3BP1 slowed down MAFLD progression by inhibiting APOC3. | https://pubmed.ncbi.nlm.nih.gov/40230220/ |
| Stress granules | Transmissible gastroenteritis virus (TGEV) | In ST cells, SG formation enhances antiviral innate immunity mediated through the retinoic acid-inducible gene I (RIG-I) signaling pathway, thereby inhibiting TGEV replication. TGEV counteracts SG formation by reducing protein levels of G3BP1 to promote its own replication, with NSP1 exhibiting the strongest inhibitory effect on SGs formation. | https://pubmed.ncbi.nlm.nih.gov/40239441/ |
| P bodies | Drug resistance | Cisplatin induces processing bodies (PB) disassembly and SG-like granule formation in A549 cells, but not in A549/DDP cells, a drug resistant NSCLC cell line, with cisplatin-induced PB disassembly occuring prior to SG-like granule formation. Also, cisplatin effectively killed A549 cells but was less toxic to A549/DDP cells, suggesting that the development of cisplatin resistance was closely related to the regulation of PBs and SG-like granules within the cancer cells. Cycloheximide (CHX) pre-treatment significantly increased the cell death of cisplatin-treated A549/DDP cells. Additionally, combination treatment with pyrvinium overcomes cisplatin resistance via mediating PB disassembly and SG-like granule assembly in cisplatin-resistant cells. | https://pubmed.ncbi.nlm.nih.gov/40247038/ |
| SEMA6A condensates | Vascular disorders | The C-terminal intrinsically disorder region (IDR) of the transmembrane semaphorin 6A (SEMA6A) forms condensates to recruit RHOA and P300, facilitating P300 phosphorylation and histone lactylation cycle. Joint CUT&Tag and scRNA-seq analyses identified Prmt5 as a target of H3K9la and H3K18la in isolated retinal endothelial cells (ECs). EC-specific deletion of Prmt5 in the early stage of revascularization suppresses a positive feedback loop of lactate production and histone lactylation, inhibiting neovascular tuft formation. Deletion of endothelial Sema6A reduced H3K9la and H3K18la at the promoter of PRMT5 and diminished its expression. The induction of histone lactylation by SEMA6A-IDR and its pro-angiogenic effect were abrogated by deletion of Prmt5. | https://pubmed.ncbi.nlm.nih.gov/40244673/ |
| CsrA foci | Bacterial infection | CsrA forms foci in enteropathogenic Escherichia coli (EPEC), with foci size increasing as cellular density progresses. Foci contain csrA, components of the RNA-degrading complex (degradosome), regulatory small RNAs, and selected mRNAs, with CsrA-RNA interactions driving foci formation. Formation of the compartment is associated with a switch between promoting and repressing virulence gene expression by CsrA. Additionally, CsrB is essential for the formation of CsrA foci in the early logarithmic growth phase, with foci formation diminished in ΔcsrB mutants. CsrA and CsrB cooperate in ler repression, and monitoring the production of the LEE1-encoded Ler and the LEE4-encoded EspD showed CsrA switches from supporting to repressing T3SS expression. | https://pubmed.ncbi.nlm.nih.gov/40268935/ |
| FUS-CREB3L2 condensates | Cancer | IDR-rich transmembrane fusion proteins (MFPs) generated by chromosomal translocations can remodel their targeted endomembrane in U2OS cells. FUS-CREB3L2 (FC), prevalent in low-grade fibromyxoid sarcoma (LGFMS), as a representative of MFPs show de novo remodeling of its targeted membranous organelles, depending on its LLPS propensity, posttranslational modification (PTM) state, and membrane topology in U2Os cells. Super-resolution long-time imaging of endoplasmic reticulum (ER) remodeling dynamics by FUS-CREB3L2 show spontaneous ER stress that hijacks the X-box-binding protein 1 (XBP1) pathway. DNA binding prevents FC liquid condensates from transiting into a gel state, leading to activation of LGFMS-specific genes. In U2OS cells, FC expression causes spontaneous ER stress and autonomous nuclear translocation, with FC-N condensates hijacking the XBP1 pathway, rewiring protein interaction networks to promote cell proliferation and migratory ability. | https://pubmed.ncbi.nlm.nih.gov/40273917/ |
| Stress granules | Prolactinoma | Cholesterol enhanced resistance of MMQ cell lines to cabergoline, and in vivo experiments revealed that a high-cholesterol diet for six weeks resulted in elevated levels of cholesterol and triglycerides in nude mice. Immunoprecipitation combined with mass spectrometry show cholesterol increased binding affinity between DRD2 and stress granule (SG) protein G3BP1. Fluorescence staining of SGs using TASG in MMQ cells, immunohistochemical staining of G3BP1 in mice tumor xenografts, and immunohistochemical staining of G3BP1 in sensitive and resistant human prolactinomas show that cholesterol promoted the formation of stress granules. Interfering with the formation of SGs by overexpressing of USP10 and using the small molecule Integrated Stress Response Inhibitor (ISRIB) reversed cholesterol’s effect on DRD2 cellular distribution and DA resistance in MMQ cells. | https://pubmed.ncbi.nlm.nih.gov/40281543/ |
| SUZ12 puncta | Duchenne muscular dystrophy (DMD) | Overexpression of lncRNA-MEG3 in C2C12 myoblasts enhanced the formation of SUZ12 puncta, whereas knockdown reduced formation. Functional assays show SUZ12 constructs lacking the IDR failed to induce H3K27me3 modification at the Fhl3 and Rnf128 promoter and couldn’t regulate their expression. Overexpression of SUZ12 rescued the ability of lncRNA-MEG3 knockdown to inhibit H3K27me3 modification, and upregulation of FHL3 and Rnf128, an effect that was abolished when the SUZ12 IDR was deleted. LncRNA-MEG3 is essential for muscle mass maintenance and regeneration in human skeletal muscle development, aging, and the mouse Duchenne muscular dystrophy (MDX) model mice, where depletion leads to muscle atrophy, mitochondrial dysfunction, and impaired regenerative capacity, while overexpression enhances muscle mass, increases oxidative muscle fiber content, and improves endurance. lncRNA-MEG3 overexpression in MDX model mice significantly alleviates musclewasting and adipose tissue infiltration. | https://pubmed.ncbi.nlm.nih.gov/40285575/ |
| PRCC fusion condensates | Translocation Renal cell carcinoma (tRCC) | Immunofluorescence of patient-derived xenografts (PDXs) harboring WT TFE3 or PRCC-TFE3 show TFE3 fusions enhance condensate formation relative to the replaced WT sequence. Measurement of reporter gene expression also reveal enhanced gene activation relative to WT TFE3. In UTSW-XP121 cells, knockdown of PRCC-TFE3 impairs gene activation and proliferation in UTSW-XP121 cells. In vitro reconstitution of condensates by adding purified PRCC-TFE3 into a nuclear extract show condensates of fusion partners partition RNA Pol II, and ChIP-seq UTSW-XP121 cell lines show PRCC TFE3 is associated with higher RNA Pol II recruitment relative to WT TFE3, and loss of PRCC-TFE3 leads to loss of RNA Pol II recruitment to target gene promoters. Fusion partner condensates interact with RNA Pol II CTD, and shared enrichment in pi and pi- interacting residues and depletion of aliphatic residues in fusions is necessary for RNA Pol II CTD partitioning and gene activation. Through review of a compiled database of oncofusions, it was found that other oncofusions from diverse cancers have the same molecular signature. Introduction of fusion-enriched amino acids in PRCC-TFE3 shRNA cells lead to gain of proliferation, migration, and invasion. | https://pubmed.ncbi.nlm.nih.gov/40286793/ |
| ASPL fusion condensates | Translocation Renal cell carcinoma (tRCC) | Immunofluorescence of patient-derived xenografts (PDXs) harboring WT TFE3 or ASPL-TFE3 fusions enhanced condensate formation relative to the replaced WT sequence. Measurement of reporter gene expression also reveal ASPL-TFE3 enhance gene activation relative to WT TFE3, and immunofluorescence of FU-UR-1 cells show ASPL-TFE3 recruit more RNA Pol II to target genes than WT TFE3. In vitro reconstitution of condensates by adding purified ASPL-TFE3 into a nuclear extract show condensates of fusion partners partition RNA Pol II. Through review of a compiled database of oncofusions, it was found that other oncofusions from diverse cancers have the same molecular signature. | https://pubmed.ncbi.nlm.nih.gov/40286793/ |
| SFPQ fusion condensates | Translocation Renal cell carcinoma (tRCC) | Immunofluorescence of patient-derived xenografts (PDXs) harboring WT TFE3 or SFPQ-TFE3 fusions enhanced condensate formation relative to the replaced WT sequence. Measurement of reporter gene expression also reveal SFPQ-TFE3 enhance gene activation relative to WT TFE3. In vitro reconstitution of condensates by adding purified SFPQ-TFE3 into a nuclear extract show condensates of fusion partners partition RNA Pol II. Through review of a compiled database of oncofusions, it was found that other oncofusions from diverse cancers have the same molecular signature. | https://pubmed.ncbi.nlm.nih.gov/40286793/ |
| A body | Neurological disorders | Image-based high-throughput screening of MCF-7 cells show that the phosphatidylinositol 3-kinase (PI3K)/AKT signaling axis meditates A-body formation during stress exposure, with AKT activation repressing glycogen synthase kinase-3 (GSK3)-mediated degradation of c-Myc. qPCR showed that inhibitors of PI3K (Pilaralisib, Torin1, Ipatasertib), mTORC1/2, and AKT reduced heat-mediated induction of rIGS16RNA without affecting GAPDH mRNA levels, observed in siRNA-mediated depletion of AKT1 as well. c-Myc accumulates in the nucleolus during heat shock to induce rIGS16RNA expression. In MCF-7 and U-87 MG cells, depletion of c-Myc using siRNA reduced both A-body formation and levels of rIGS16RNA expression under heat shock conditions without affecting GAPDH, suggesting that c-Myc activity is involved in this amyloid aggregation pathway. | https://pubmed.ncbi.nlm.nih.gov/40272983/ |
| Nucleolus | Neurological disorders | In MCF-7 cells, c-Myc accumulates in the nucleolus during heat shock to induce rIGS16RNA expression. qPCR shows this occurs prior to the heat-induced expression of rIGS16RNA, highlighting that nucleolar localization is not mediated by rIGSRNA itself. | https://pubmed.ncbi.nlm.nih.gov/40272983/ |
| Stress granules | Neurodegeneration | Dipeptide repeats poly-PR and poly-GR, formed through translation of C9ORF72 repeat expansion, induce formation of and localize to stress granules, to impair translation. Expression of GR and PR peptides in HeLa and U2-OS cells promoted spontaneous assembly of stress granules. FRAP experiments and live cell imaging demonstrate that the GR and PR-containing stress granules exhibit decreased dynamics compared to SGs induced by oxidative stress. poly-GR and poly-PR peptides promoted formation of hnRNPA1 and TIA-1 droplets in vitro. | https://pubmed.ncbi.nlm.nih.gov/27768896/ |
| Stress granules | Traumatic brain injury (TBI)|Amyotrophic lateral sclerosis (ALS) | Traumatic brain injury (TBI) in Drosphilla induces formation of stress granules (SGs) that co-localize with phosphorylated Ref(2)P and transiently recruit TBPH. Traumatic injury in Drosophila models of ALS (expressing FUS mutants or C9ORF72 repeat expansions) increase levels of Ref(2)P (p62 homologue), enhance animal mortality and locomotive dysfunction. | https://pubmed.ncbi.nlm.nih.gov/29432563/ |
| Transcriptional condensates | Cancer | ZHX2 forms nuclear condensates in MCF7 cells, T47D cells, and tumors from MDA-MB-231 xenograft tumor-bearing mice and breast cancer patients, with the proline-rich ZHX2 IDR mediating hypoxia-induced LLPS. Fluorescence images, co-localization analysis, and immunoblots of lysates of MDA-MB-231 cells show that ZHX2 forms transcriptionally active condensates in hypoxia. Integration of data from ChIP-seq, RNA-seq, and an assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) reveal hypoxia-induced LLPS of ZHX2 activates a cluster of oncogene transcription that is enriched by metastatic genes distinct from the targets of hypoxia-inducible factor (HIF). LLPS-enhanced phosphorylation of ZHX2 at S625 and S628 partitions CTCF into condensates in MDA-MB-231 cells, and non-invasive bioluminescence imaging in mice and ex vivo imaging of lung tissue showed that ZHX2 depletion inhibited breast cancer lung colonization and outgrowth. | https://pubmed.ncbi.nlm.nih.gov/40185097/ |
| ca-circSCN8A/FUS/EP300 condensates | Pulmonary hypertension (PH) | ca-circSCN8A was identified via bioinformatics, Sanger sequencing, and RNase R digestion and confirmed to be upregulated in pulmonary hypertension (PH) via functional gain and loss assays. Its overexpression promoted hypoxia-induced ferroptosis in human pulmonary arterial smooth muscle cells and a PH mouse model. Under hypoxic conditions, ca-circSCN8A recruited EP300 to facilitate the lactylation of FUS, triggering the formation of a ca-circSCN8A/FUS/EP300 complex via LLPS. Molecular mechanisms were probed via FRAP, ChIRP-qPCR, malondialdehyde, reduced glutathione, and glutathione, and showed that LLPS maintained the stability of the R-loop formed by ca-circSCN8A and ferroptosis-related gene SLC7A11 promoter that inhibits its transcription, further result in the disruption of the redox homeostasis and causing ferroptosis in human pulmonary arterial smooth muscle cells. | https://pubmed.ncbi.nlm.nih.gov/40384439/ |
| Stress granules | Dengue virus (DENV) | G3BP1 formed cytoplasmic SGs in uninfected arsenite-treated HEK 293T cells, but no SGs were detected in DENV-2-infected cells. DENV-2 dsRNA, DDX17, and G3BP1 were found to co-localize in infected cells, forming perinuclear aggregates. Western blot results confirmed that DDX17 interacts with G3BP1. Furthermore, DENV-2 mRNA levels were significantly increased in the DDX17 and G3BP1 co-transfection group at 24 hours post-infection, and plaque assays of the culture supernatant confirmed that viral titer was higher in the co-transfection group compared to the control. | https://pubmed.ncbi.nlm.nih.gov/40383034/ |
| RXRγ condensates | Small-cell lung cancer (SCLC) | Retinoid X receptor γ (RXRγ) is uniquely overexpressed in chemo-resistant Small-cell lung cancer (SCLC) tumors, serving as a factor driving chemoresistance in SCLC, and RXRγ overexpression promoted the growth and metastasis of H146 xenograft tumors in mice. RXRγ forms droplets with LSD1 in the nucleus which enhances RXRγ-mediated gene transcription activity and reprograms gene expression, promoting tumor stemness and metastasis, and eventually driving SCLC chemoresistance. Additionally, more and larger nuclear puncta of LSD1-RXRγ were observed in in chemo-resistant SCLC tumors. RXRγ antagonist HX-531 disrupts RXRγ–LSD1 interaction, reduces binding to the target gene locus, and suppresses the expression of the RXRγ target gene network. In preclinical SCLC models, RXRγ antagonists strongly suppress tumor growth and metastasis and restore SCLC vulnerability to chemotherapy. | https://pubmed.ncbi.nlm.nih.gov/40392852/ |
| YAP condensates | Hepatocellular carcinoma (HCC) | Cancer-associated fibroblasts (CAFs) isolated from tumor tissue samples of Hepatocellular carcinoma (HCC patients) were found to promote cancer stem cell (CSC) stemness capacity via extracellular vesicles (EVs). Additionally, transwell migration and cloning formation assays reveal NEAT1 overexpression significantly increased the migration and invasion of MHCC-97 H and SNU-398 cells, and si-NEAT1 treated CAFs and si-NEAT1 CAF-EVs exhibit a decrease in migration. RNA pull-down and RNA immunoprecipitation (RIP) assays confirm an interaction between NEAT1 and YAP. YAP-GFP in cellular condensates recovers from photobleaching (FRAP) within minutes. Investigation of the function of YAP LLPS found that PEG400 stimulation enhanced cell proliferation, invation and migration of MHCC-97 H and SNU398 cells, whereas Hex-1,6 had the opposite effect. EVs derived from CAFs facilitated the phase separation of YAP, and formation of YAP droplets was dependent on the concentration of CAFs-EVs and were no longer observed upon silencing of NEAT1. Injection of MHCC-97 H, and SNU-398 cells into male BALB/c nude mice found the mean volume of tumor nodules in the sh-NEAT1 group was significantly lower, and immunohistochemical staining and Western blot showed interfering the expression of NEAT1 reduced the expression of Nanog and YAP, indicating NEAT1 regulates tumor stemness capacity in HCC in vivo. Overall, NEAT1 promoted HCC carcinogenesis and CSC induction by facilitating the LLPS of the YAP protein. | https://pubmed.ncbi.nlm.nih.gov/40413530/ |
| Stress granules | Amyotrophic lateral sclerosis (ALS)|Frontotemporal dementia (FTD) | In a sporadic type B frontotemporal dementia (FTD) patient, mildly affected neurons retained nuclear TDP-43 but exhibited small cytoplasmic puncta displaying phosphorylated TDP-43 and intra-condensate demixing within stress granules. Severely affected neurons lost nuclear TDP-43 and displayed large cytoplasmic phosphorylated aggregates interacting with stress granules and showing demixing. In cells showing arsenite-induced demixing, FRAP reveals that TDP-43 dynamics gradually slowed while remaining dynamic under puromycin, indicating TDP-43 hardening during demixing. These demixed puncta were co-stained with HSPB1 and exhibited ubiquitination, linking demixing to TDP-43 aggregation in iPS-motor neurons. In rNLS8 mice, TDP-43 aggregation was absent without hTDP-43ΔNLS induction or in unaffected neurons with induction, and showed either strong colocalization of TDP-43 with stress granules or a distinct demixing pattern. In control postmortem brain samples TDP-43 was predominantly nuclear with no phosphorylated aggregates. | https://pubmed.ncbi.nlm.nih.gov/40412392/ |
| p62 bodies | Liver damage | p62 knock-in mice with diminished KEAP1 binding or entirely lacking KEAP1 retention were crossbred with Atg7flox/flox;Alb-Cre mice. Hematoxylin, eosin, and CK19 staining show that the level of KEAP1 retention in p62 bodies determines the severity of liver pathology in the hepatocyte-specific Atg7-knockout mice with different p62 mutations. Immunofluorescence of primary cultured hepatocytes--accumulating p62 bodies that sequester KEAP1, reduced KEAP1 retention in p62 bodies, and accumulating p62 bodies with minimal KEAP1 retention, subsequently revealing that KEAP1 is sequestered into p62 bodies based on its binding affinity to p62. Immunoblot analysis of these mouse models show autophagy degrades KEAP1 incorporated into p62 bodies but not than cytosolic KEAP1. Furthermore, quantitative real-time PCR analysis confirmed that NRF2 target genes were upregulated in Atg7-KO mouse livers but suppressed in a stepwise manner when wild-type p62 was replaced with p62S351A or p62T352A, with Sqstm1 expression upregulated in Atg7-KO livers and reversed by substituting wild-type p62 with either mutant. Immunoblot analysis showed significant increases in NRF2 target proteins, overall demonstrating transcriptomic changes caused by Atg7 deficiency involving the NRF2 pathway are dependent on the binding affinity of KEAP1 to p62 bodies, with cytoplasmic proteome profiling of the mice livers confirming proteome-wide changes driven by KEAP1-p62 binding. | https://pubmed.ncbi.nlm.nih.gov/40437287/ |
| Nucleolus | Ribosomopathy | HNRNPK protein levels were found to be elevated in bone marrow samples of patients with ribosomopathies that cause bone marrow failure. HnrnpkTg mice had quicker exhaustion rates, showed bone marrow failure, lifespan reduction and an aging phenotype, partially rescued by p53-haploinsufficiency. HNRNPK-overexpressing mouse models showed a significant enrichment of a set of downregulated genes and proteins involved in ribosome biogenesis (RiBi) and rRNA processing, with nucleolin immunolabeling and electron microscopy analysis showing an increased number of nucleoli, as well as smaller nucleoli. Additionally, nucleolar alterations including segregation of components at the nucleolar periphery, abnormal accumulations of the granular component, nucleolar lobulation and gramentation, and formation of prominent masses of repressive heterochromatin. HNRNPK overexpression disrupted normal ribosomal biogenesis, leading to altered rRNA processing and aberrant rRNA precursor levels, and nucleolar stress derived from HNRNPK overexpression promoted cell cycle arrest and senescence via p53. | https://pubmed.ncbi.nlm.nih.gov/40338663/ |
| αSyn inclusions | Parkinson's disease (PD) | Knockdown of TAX1BP1 or ADAMTS19 through small interfering RNAs (siRNAs) resulted in a significant increase in the number and decrease in the size of αSyn inclusions in M17D cell lines. FRAP experiments show that αSyn inclusions are partially dynamic, and knockdown of ADAMTS19, but not TAX1BP1 result in a decrease of the immobile fraction (Fi) of αSyn inclusions. Treatment with oleic acid in resulted in enlarged αSyn inclusions with a “Swiss-cheese” appearance, but knockdown of TAX1BP1 resulted in large lipid droplets (LDs) surrounded by a thin rim of αSyn--“hollow” inclusions. In the complete absence of αSyn, knockdown of TAX1BP1 and ADAMTS19 had no effect on LDs. The mutations in the 3K (artificial amplification of the E46K mutation) αSyn protein increase its propensity to phase separate and associate with phospholipids in vitro and in M17D cells. Additionally, MD simulations confirm the importance of the lysines at residues 35, 46, and 61 in 3K αSyn for its conformation and interaction with lipids. 3K M17D cells treated with Chloroquine showed a significant decrease in the number of αSyn inclusions per cell, including after knocking down TAX1BP1 or ADAMTS19. Oleic acid treatment prevented the removal of Swiss-cheese lipid-rich αSyn inclusions but not of the solid inclusions. Weighted gene co-expression network analyses on gene expression data show TAX1BP1 and ADAMTS19 integrate within genetic networks carrying an increased burden of rare variants in patients with PD. | https://pubmed.ncbi.nlm.nih.gov/40317721/ |
| ViPR bodies | Adenovirus | In adenovirus-infected U2OS cells, NPM1 relocalizes from the nucleolus to nuclear viral protein VII to viral-induced post replication (ViPR) bodies to promote viral genome packing. NPM1 is highly mobile in ViPRs, with FRAP recovery times on the order of seconds. Protein VII forms heterotypic LLPS with NPM1 in vitro. | https://pubmed.ncbi.nlm.nih.gov/39162498/ |
| Stress granules | SARS-CoV-2 | SARS-CoV-2 infection leads to increased expression of HDAC6 in infected VERO, A549 and Calu-3 cell lines, and promoted viral replication. SARS-CoV-2 infection promotes interaction between G3BP1 and HDAC6, and between SARS-CoV-2 N protein and HDAC6, as measured by immunoprecipitation. Interaction of HDAC6 with the N protein stabilizes HDAC6, as a mechanism of increased expression. N protein expression inhibits formation stress granule induction in response to oxidative stress. | https://pubmed.ncbi.nlm.nih.gov/39135075/ |
| USP49 condensates | Radioresistance|Esophageal squamous cell carcinoma (ESCC) | Elevated USP49 expression was related to a poorer prognosis in esophageal squamous cell carcinoma (ESCC) patients receiving chemoradiotherapy, and found to facilitate the proliferation and radioresistance of ESCC cell lines. The ataxia telangiectasia mutated (ATM) promotes LLPS of USP49 and its recruitment at DNA double -strand breaks (DSBs) upon ionizing radiation (IR) exposure, coordinating the modulation of homologous recombination (HR) repair. Olaparib induced more pronounced LLPS of USP49 compared to untreated cells. Additionally, USP49 interacts with the DNA-binding domain B of RPA70 and catalyzes its deubiquitination through its USP domain, thereby enhancing HR and radioresistance. USP49 deubiquitinates and recruits RPA70 to DSBs along with Rad51, thereby promoting HR repair. Furthermore, USP49 is transcriptionally upregulated by p53 following IR exposure. | https://pubmed.ncbi.nlm.nih.gov/40460957/ |
| PMMoV CP condensates | Pepper mild mottle virus (PMMoV) | Pepper mild mottle virus (PMMoV) CP promoted LLPS by interacting with I3QHX5 to facilitate viral replication, and silencing of I3QHX5 attenuates the infection of PMMoV in N. benthamiana. ABPP technology, Western blot, and microscale thermophoresis show that PMMoV CP was the target protein of purine morpholine nucleoside analogue C1, with confocal microscopy and RT-qPCR further revealing that Tyr13 is the key amino acid site for C1 action and could severely inhibit virus infection. Additionally, bimolecular fluorescence complementation demonstrates that PMMoV CP Y13A does not exhibit LLPS. Overall, PMMoV CP may drive LLPS by interacting with I3QHX5 to facilitate viral replication, while PMMoV CPY13A does not exhibit LLPS. C1 may specifically targeted PMMoV CPY13A to inhibit LLPS, thereby suppressing viral replication. | https://pubmed.ncbi.nlm.nih.gov/40460937/ |
| Stress granules | Hepatocellular carcinoma (HCC) | Atypical serine/threonine kinase RIOK1 is highly expressed in hepatocellular carcinoma (HCC), linked to poor prognosis, and transcriptionally activated by NRF2 under various stress conditions, and RIOK1-positive stress granules (SGs) are found in donafenib-resistant tumors from patients with HCC. In vitro and in HCCLM3 cells, RIOK1 undergoes LLPS and drives SG assembly via IGF2BP1–G3BP1 interactions, and RIOK1 LLPS inhibits PTEN translation by hijacking PTEN mRNA within SGs. Additionally, RIOK1-induced LLPS impairs HCC sensitivity to tyrosine kinase inhibitors (TKIs). Silencing RIOK1 inhibits pentose phosphate pathway and tumor growth in liver tumor mouse models. Small-molecule inhibitor chidamide downregulates RIOK1 and enhances TKI efficacy. | https://pubmed.ncbi.nlm.nih.gov/40467995/ |
| PKA Riβ condensates | Neurodegeneration | In HEK293T cells, RIβ mutant L50R linked to neuronal loss and parkinsonism driven by a PKA mutation (NLPD-PKA) or RIβ mutant R335W linked to Marbach-Schaaf neurodevelopmental syndrome (MASNS) produce aberrant condensates that trap the PKA catalytic subunit within a gel-like matrix or cAMP-insensitive holoenzyme complex, respectively. Measurements of dendritic arbors of mixed primary hippocampal and cortical neurons from mice reveal neurite outgrowth was significantly decreased in cells expressing RIβ-L50R. RIβ immunofluorescence staining shows formation of puncta in PC12 cells, and purified RIβ protein also forms assemblies in vitro at concentrations as low as 4 μM. FRAP measurements in HEK293T cells further reveal that RIβ LLPS require both the D/D domain and N3A-A motif. Furthermore, the presence of Cα recovers liquid-like condensate formation in RIβ-ΔLinker, and smAKAP recruitment inhibits RIβ condensates. | https://pubmed.ncbi.nlm.nih.gov/40512625/ |
| TRIM24 condensates | T-cell lymphoma (TCL) | TRIM24 is overexpressed in peripheral T-cell lymphoma (TCL) samples and correlates with suppressed glucocorticoid receptor (GR) signaling and poor therapeutic response. Transcriptomic and chromatin profiling show TRIM24 deficiency delays TCL progression in murine models, suppresses cell proliferation, and enhances glucocorticoid sensitivity by restoring GR transcriptional activity. TRIM24 interacts with FUS and promotes its LLPS with GR, leading to impaired GR activity and heightened GC resistance. These findings support a “double-check” mechanism in which TRIM24 regulates nuclear GR function through FUS-facilitated LLPS. | https://pubmed.ncbi.nlm.nih.gov/40513312/ |
| FfSR condensates | Rice bakanae disease | Deletion of FfSR results in reduced ergosterol levels, increasing the susceptibility of F. fujikuroi to azole fungicides. Azole-induced ergosterol depletion promotes FfSR phase separation, facilitating its binding to cis-elements at target promoters, subsequently activating the expression of ergosterol biosynthesis genes. High levels of ergosterol bind to FfSR, inhibiting phase separation and transcriptional activation. Additionally, natamycin was found to be a direct inhibitor of FfSR, suppressing its phase separation and transcriptional capability. | https://pubmed.ncbi.nlm.nih.gov/40514608/ |
| WNK body condensates | Hypokalemia | WNK bodies were present in human kidney samples, with an inverse correlation between decreasing serum potassium and increasing WNK body abundance, where WNK bodies accumulate as plasma potassium falls below 4.0 mmol/L. In male mice kidneys, WNK body size increased exponentially as blood potassium levels progressively decreased below 4 mmol/L, and WNK bodies were not visible in male mice with a plasma potassium greater than 4.0 mmol/L. During potassium deficiency, kidney specific (KS)-WNK1 drives WNK body formation, amplifies NaCl cotransporter (NCC) phosphoactivation, and increases WNK4-SPAK/OSR1 pathway expression. Additionally, KS-WNK1 5Q mutation alters WNK body formation and function during K+ restriction, resulting in mislocalization of the WNKSPAK/OSR1 pathway and low NCC activity. Genetically modified mice with targeted condensate disruption trap the WNK pathway, causing renal salt wasting that is more pronounced in females. | https://pubmed.ncbi.nlm.nih.gov/40493421/ |
| Stress granules | Doxorubicin-induced cardiotoxicity | In H9c2 cardiomyocytes DEAD-box RNA helicase 3 X-linked (DDX3X), mitochondrial antiviral signaling (MAVS) and stress granules (SGs) were present at lower levels in H9c2 cardiomyocytes treated with doxorubicin (Dox) compared to untreated control cells. Pretreatment with arsenite, a SG inducer, alleviated myocardial injury caused by Dox. Anisomycin, an SG inhibitor, increased cardiomyocyte apoptosis induced by Dox. In addition, both DDX3X knockdown and pretreatment with RK-33 (a DDX3X pharmacological inhibitor) decreased SG expression, whereas DDX3X overexpression promoted SG generation. MAVS knockdown inhibited SG assembly and reduced the expression of the anti-apoptotic inhibitor Bcl2, and MAVS was influenced by DDX3X, thereby serving as a connector between DDX3X and SGs. | https://pubmed.ncbi.nlm.nih.gov/40576139/ |
| β-catenin condensates | Liver cancer | Abroquinone (AQ) treatment inhibited the hyperactivated Wnt/β-catenin pathway in hepatoma carcinoma cell (HCC) mouse models and stimulated T-cell-mediated immunity. scRNA-seq analysis of a cluster of tumor cells show downregulated expression of β-catenin, Cyclin D1, and Wnt5a, and upregulation in the proportion of CD8 T cells. Single-sample geneset enrichment analysis reveals suppression of the Wnt/β-catenin signaling and cell cycle pathways. In Hep3B HCC cells, Rosmarinol quinone (RQ) treatments induced the formation of spherical β-catenin condensates whereas under control conditions, β-catenin exhibits diffuse intracellular localization. Condensate number increased with 5μM RQ treatment compared to 2.5μM RQ treatment. Quantification of β-catenin distribution via FRAP show treatment with 5μM RQ led to a reduction in intranuclear β-catenin percentage from 53.7 to 20.5%, and treatment with 10μM RQ significantly suppressed the nuclear translocation of β-catenin, forcing β-catenin into cytoplasmic condensates. Decreased protein levels of β-catenin, active β-catenin, c-myc and Cyclin D1 were observed in Wnt-hyperactivated HCC cell lines Hep3B, HepG2, and Hepa1-6 following RQ treatment. Additionally, cell viability assays show dose-dependent growth inhibitions induced by RQ in ten HCC cell lines while showing minimal cytotoxicity towards normal liver cell lines, primary stem cells, and immune cell lines. | https://pubmed.ncbi.nlm.nih.gov/40593772/ |
| P bodies | Myotonic dystrophy type 1 (DM1) | MBNL1 and CUGBP1 colocalize with P bodies in Myotonic dystrophy patient-derived human lens epithelial cells (HLECs) and HeLa cell models of DM1. P body-localized MBNL1 has a slower recovery time in FRAP in DM1 HLECs compared to control HLECs. Stress granules (SGs) in HeLa cells expressing CTG expansion contain less poly(A) RNA and show altered docking events with P bodies. | https://pubmed.ncbi.nlm.nih.gov/35642886/ |
| ACK1 condensates | Lung squamous cell carcinoma (LUSC) | Analysis of the TCGA PanCancer Atlas found ACK1 is amplified and highly expressed in lung squamous cell carcinoma (LUSC). FRAP showed ACK1 undergoes LLPS with fluid-like, dynamic characteristics, and photobleaching analysis demonstrated rapid fluorescence recovery within minutes. Exogenous ACK1-GFP localizes to condensates in H2170 cells, and subcellular localization analysis in HeLa cells show mutants lacking the IDR1 region (ΔIDR1) have a diffuse distribution, whereas mutants lacking IDR2, IDR3, IDR4, or IDR5 showed a punctate distribution similar to WT. ACK1 overexpression enhanced phosphorylation of exogenous STAT5 in H2170 cells, and only constitutively active (CA) mutants induced phosphorylation of STAT5, whereas kinase-dead (KD) mutants did not. Additionally, treatment with ACK1 kinase activity inhibitor AIM-1000 abolished p-STAT5 signal. An ACK1 IDRmut (ΔIDR1-5) defective in condensate formation failed to promote both endogenous and exogeneous p-STAT5 signaling, but exhibited Tyr284 autophosphorylation levels comparable to ACK1, revealing loss of p-STAT5 signaling was not due to a lack of ACK1 kinase activity. Immunofluoresence showed that only ACK1 WT, and not IDRmut promoted the nuclear localization of STAT5, and RNA-seq found that a STAT5 target gene set was upregulated in cells overexpressing ACK1 WT, but not IDRmut. Colony formation assays and scratch wound assays show ACK1 deletion significantly reduced the number of colonies and markedly suppressed cell migration, respectively. | https://pubmed.ncbi.nlm.nih.gov/40581662/ |
| Stress granules | Amyotrophic Lateral Sclerosis (ALS) | Amyotrophic Lateral Sclerosis (ALS) | https://pubmed.ncbi.nlm.nih.gov/40622676/ |
| WDR3 condensates | Osteosarcoma | In transfected U2-OS cells and xenograft mice, the downregulation of WDR3 significantly inhibited the malignant progression of osteosarcoma. WDR3 forms droplets in U2-OS cells, and and restores the fluorescence intensity of WDR3 condensates with liquid-like behavior after photobleaching within 60 seconds. Additionally, a mutation in its IDR impairs the phase separation ability of WDR3, whereas the fusion with hnRNAP1 IDR rescued the phase separation abnormality. Treatment with Nilotinib improves the progression of osteosarcoma while inhibiting the production of WDR3 condensates. | https://pubmed.ncbi.nlm.nih.gov/40646517/ |
| Stress granules | Allergic rhinitis | Immunofluorescence imaging of G3BP1 in the nasal mucosa from OVA-induced allergic rhinitis (AR) mice shows stress granules (SGs) are assembled in the macrophages of the nasal mucosa during AR. G3bp1f/f and G3bp1mac-/- mice administered ovalbumin (OVA) or house dust mites (HDM) to trigger AR symptoms reveal SG assembly in macrophages promotes AR progression, and transcription analysis of isolated BMDMs show that SG assembly in macrophages impairs efferocytosis. RNA-seq of BMDMs post HDM stimulation and G3BP1 RNA immunoprecipitation sequencing in RAW 264.7 macrophages transfected with Flag-tagged G3BP1 found SG assembly in macrophages sequesters Lrp1 mRNA to suppress its translation. Polysome profiling and protein degradation assays in macrophages further show internal m7G-modified Lrp1 mRNA is shuttled by QKI7 into SGs, thus repressing its translation. Lrp1mac-/- mice administered with OVA displayed more severe AR symptoms with increased sneezing and scratching counts compared to G3bp1mac-/-. Efferocytosis assays determined that LRP1 deficiency disturbed macrophage phagocytotic ability to clear apoptotic cells. BMDMs treated with G3BP1 inhibitor G3Ia followed by administration of HDM showed significantly lowered SG assembly in a dose dependent manner. Additionally, oxidative stress caused more severe AR symptoms, while G3Ia treatment effectively alleviated AR progression, symptoms, and mucosa damage, and rescued the defect of macrophage efferocytosis under stress and restored LRP1 expression at the protein level. Furthermore, SGs were assembled in the macrophages of nasal mucosa from AR patients, but not in the healthy control. | https://pubmed.ncbi.nlm.nih.gov/40595582/ |
| YTHDC1 condensates | Renal cell carcinoma (RCC) | Human renal cell carcinoma (RCC) tissues and cells showed elevated global Kla levels which promoted RCC malignancy. Lactylome analysis of human RCC cells under hypoxia-mimicking conditions revealed YTHDC1 is modified by Kla at K82. YTHDC1-K82la, mediated by p300 under hypoxia, promoted RCC malignancy and increased YTHDC1 phase separation, leading to the expansion of nuclear condensates and safeguarding oncogenic transcripts BCL2 and E2F2 from degradation by the PAXT-exosome complex in human RCC cells. | https://pubmed.ncbi.nlm.nih.gov/40680722/ |
| FGFBP1 condensates | Acute liver injury (ALI) | Integrated analysis of RNA-seq and ATAC-seq following acute liver injury (ALI) in mice found that ALI induces glucocorticoid (GC)-mediated transrepression of Fgf6 in skeletal muscle. Skeletal muscle glucocorticoid receptor (GR) activation after ALI induces muscle loss and is beneficial to ALI repair, whereas knockdown exacerbates ALI and suppresses liver regeneration. FGF6 acts downstream of GCs to mediate the muscle metabolic response to ALI and influence liver regeneration--FGF6 overexpression in the skeletal muscle decelerated liver regeneration after partial (2/3) hepatectomy (PHx), and FGF6 knockout protects from PHx and acetaminophen (APAP)-induced ALI and enhances liver regeneration. Intramuscular injection of FGF6-neutralizing antibody (FGF6Ab) rescues the detrimental effects induced by GR knockdown. RNA-seq data of muscle with FGF6 overexpression reveals that FGF6 regulates FGFBP1 expression through extracellular signal regulated kinase-activating transcription factor 3 signaling. Luciferase and ChIP assays confirmed the activation of ATF3 to Fgfbp1 transcription, while these effects were inhibited by rFGF6. In AML12 cells, immunofluorescence assays showed FGFBP1 was distributed in a punctate pattern and colocalized with FGF5 in areas with APAP-induced damage. Fluorescence signals recovered 3 mins after photobleaching, and quantification of FRAP micrographs confirms LLPS. Purified protein colocalization assays on AML12 cell surfaces via confocal microscopy showed FGF5 and FGF5s compete for binding to FGFBP1, and the addition of rFGF5s diminished FGF5-FGFBP1 condensation in a dose-dependent manner. FGFBP1, FGF5, and FGF5s combination therapy on liver regeneration of ALI mice revealed that rFGFBP1 + rFGF5 treatment largely promoted liver regeneration, whereas these benefits were blunted upon treatment with rFGF5s. Clinical studies show patients with high serum FGFBP1 recover faster after ALI, with a close correlation between serum FGFBP1 with the decline rate of ALT and AST serum liver enzyme levels. | https://pubmed.ncbi.nlm.nih.gov/40685360/ |
| LINC00599 condensates | Pulmonary hypertension | The expression of LINC00599 is upregulated in the medial layer of pulmonary arteries in a murine pulmonary hypertension (PH) model and hypoxic pulmonary arterial smooth muscle cells (PASMCs). Administration of lentivirus-mediated shRNA targeting LINC00599 reverses hypoxic PH in murine models. LINC00599 promotes PASMC proliferation by modulating stress granule formation through m6A modification and facilitating LLPS with MYH9. Furthermore, its expression is driven by a super-enhancer mediated by the transcription factor ZNF263. | https://pubmed.ncbi.nlm.nih.gov/40693377/ |
| Stress granules | Atherosclerosis | Western blot, PCR and immunofluorescence staining of the aorta of ApoE−/− mice fed with a high-fat diet (HFD) for 12w shows expression levels of stress granule (SG) core proteins G3BP1/2 and HDAC11 were increased, and endothelial-to-mesenchymal transition (EndMT) occurred. Additionally, treatment of HUVECs with H2O2 resulted in SG formation, HDAC11 upregulation, and EndMT occurrence. Knockdown of HDAC11 by siRNA significantly attenuated SG formation and EndMT activation in HUVECs induced by H2O2, while silencing of HDAC11 suppressed H2O2-induced EndMT activation in HUVECs. Suppression of SG formation facilitated the expression of endothelial markers, but decreased the levels of mesenchymal cell markers. Overall, HDAC11 may regulate SG formation to promote EndMT in atherosclerosis. | https://pubmed.ncbi.nlm.nih.gov/40692017/ |
| ATRX condensates | Alpha-thalassemia X-linked intellectual disability (ATR-X) syndrome | Immunostaining of in human neural progenitor cells (hNPCs) reveals that ATRX droplets compartmentalize and concentrate transcriptional activators P300, CHD7, BRD4, and MED1. Cleavage under targets and release using nuclease (CUT&RUN) assays found that ATRX binds to enhancers and regulates neural differentiation genes. FRAP experiments on ATRX-KO HEK293T cells expressing various EGFP-ATRX constructs confirmed that EGFP-ATRX, EGFP-IDR2full and EGFP-IDR2Cterm exhibited properties of liquid-like condensates, with apparent diffusion coefficients of ~0.11 ± 0.03 μm2/s, whereas EGFP-IDR2Nterm did not. To assess whether phase separation mediates ATRX localization to SEs, CUT&RUN assays were conducted in 1,6-Hex-treated hNPCs and found that although 1,6-Hex treatment reduced ATRX occupancy in ATRX-binding regions in a concentration-dependent manner and altered its genome distribution, the change in ATRX binding was locus-specific. Transcriptome analysis of 1,6-Hex-treated hNPCs revealed significant downregulation of the “brain development” pathway as one of the most affected biological processes, based on comprehensive GO analysis of the differentially expressed genes. LLPS-resistant mutants of ATRX (ΔEx15) cause dysregulated neural differentiation in hiPSCs. Cerebral organoids derived from ΔEx15 hiPSCs displayed distinct morphological abnormalities, including poorly defined tubular structures and a smaller size relative to WT organoids. | https://pubmed.ncbi.nlm.nih.gov/40659667/ |
| Nucleolus | Castration-resistant prostate cancer (CRPC)| neuro-endocrine prostate cancer (NEPC) | Immunohistochemistry (IHC) staining of NPM1 and FBL in prostate cancer (PCa) patient specimens reveal that the expression of NPM1 and FBL was enhanced in aggressive castration-resistant PCa (CRPC) and neuro-endocrine PCa (NEPC) patient specimens compared to hormone-naïve PCa (HNPC) patient specimens. Additionally, the expression of NPM1 was enhanced in high-Gleason score PCa compared to low-Gleason score PCa. Immunofluorescence of LNCaP, PC3, castration-resistant LNCaP C4-2, 22RV1, noncancerous BPH-1 and noncancerous PNT1B cells show NPM1 and FBL localize in the nucleolus.Additionally, in LNCaP C4-2, 22Rv1, LNCaP, and noncancerous PNT1B cells, NPM1 silencing fragmented the nucleoli and FBL silencing condensed the nucleoli. Cell migration and invasion assays show silencing NPM1 or FBL significantly reduced the proliferation, migration, and invasion of LNCaP C4-2, 22Rv1, and LNCaP cells, but not noncancerous PNT1B cells. | https://pubmed.ncbi.nlm.nih.gov/40705480/ |
| ZHX2 condensates | Diffuse large B-cell lymphoma (DLBCL) | Immunohistochemistry (IHC) staining of patient tonsil tissue showed elevated expression of ZHX2 in both germinal center B-cell (GCB) diffuse large B-cell lymphoma (DLBCL), and non-GCB DLBCL. Additionally, RT-qPCR and Western blot show elevated mRNA and protein levels, respectively. Knockdown (KD) of ZHX2 in SU-DHL-4, DB, and U2932 cells significantly inhibited proliferation by CCK-8 and colony formation assays, while overexpression enhanced proliferation. Additionally, DLBCL xenograft mouse models show reduced tumor size and volume, and reduction of tumor infiltration and proliferation upon KD of ZHX2. FRAP revealed ZHX2 forms droplets in HEK293T cells, with fluorescence recovery reached up to ~80% within 60s. Additionally, endogenous ZHX2 particles were observed in the DLBCL primary cells, SU-DHL-4, DB and U2932 cell lines. ZHX2ΔHD3 resulted in loss of LLPS and did not promote growth of SU-DHL-4, DB, and U2932 cells, and significantly inhibited the promotive effect of ZHX2 on tumor growth in mice. ChIP-seq, RNA-seq, and dual-luciferase assays further show that the LLPS of ZHX2 repressed ferroptosis and cell proliferation by activating SLC3A2. Furthermore, nanoparticle tumor-targeted delivery of ZHX2-siRNA attenuates DLBCL progress in SU-DHL-4 xenograft mouse models via downregulation of ZHX2 and SLC3A2 expression and promotion of ferroptosis. | https://pubmed.ncbi.nlm.nih.gov/40730912/ |
| CgSCE3 condensates | Anthracnose | CgSCE3 phase separation was shown to be essential for thermotolerance and pathogenicity of Colletotrichum gloeosporioides, a major pathogen responsible for anthracnose, which causes significant economic losses in rubber tree and other tropical crops. CgSCE3 was also found to participate in the regulation of CgPab1-mediated stress granule formation during heat shock. | https://pubmed.ncbi.nlm.nih.gov/40716138/ |
| GPS2-LATS1 condensates | Colorectal cancer (CRC) | GPS2 is overexpressed in colorectal cancer (CRC) and is correlated with aggressive phenotypes. Additionally, GPS2 depletion inhibits tumor growth in cellulo and in vivo, and its overexpression accelerates malignancy. GPS2 undergoes liquid-liquid phase separation (LLPS) in CRC cells, and interacts with the C-terminal kinase domain of LATS1, inducing LLPS of LATS1 and suppressing its activity. LATS1 inactivation releases YAP, amplifying SREBP1/PPARα-driven lipid metabolism. Rescue experiments show YAP reconstitution restores SREBP1 nuclear translocation and PPARα transcription upon GPS2 loss. | https://pubmed.ncbi.nlm.nih.gov/40715488/ |
| Stress granules | Ovarian aging | NCOA7 deletion accelerates oxidative stress-related cellular senescence, ovarian aging and fecundity decline in mice. NCOA7 partitions into the stress granule containing G3BP1–V-ATPase and facilitates autophagic degradation of stress granules to relieve stress. Boosting granulophagy with rapamycin or lipid nanoparticle-based mRNA delivery of NCOA7 accelerates stress granule clearance, alleviating cellular senescence in human granulosa cells and delaying ovarian aging in mice. | https://pubmed.ncbi.nlm.nih.gov/40745099/ |
| 53BP1 Mitotic Stress Body (MSBs) | Cancer | Western blot analysis and detection of cleaved caspase-3 as marker of apoptosis by automated high-content microscopy in MCF7 and U2OS cells show CNOT1 suppresses p53-p21 signaling and limits apoptosis in a 53BP1-dependent manner. Additionally, colony formation assays reveal CNOT1 depletion impairs proliferation, induces apoptosis, and causes cancer cell death. Interaction analysis by co-immunoprecipitation, Western blot analysis, high-content microscopy-derived analysis of chromatin-bound 53BP1, and gene expression analysis by real-time qPCR found that CNOT1 interacts with 53BP1 and impacts its nuclear dynamics, and CNOT1 loss upregulates p53 target gene expression. optoDroplets assays reveal the mobility of 53BP1 droplets appeared reduced, and more pre-formed condensates were observed upon CNOT1 depletion. CNOT1 loss suppresses cytoplasmic aggregation of mutant p53, restoring nuclear localization and functionality of p53 mutants frequently found in cancer. | https://pubmed.ncbi.nlm.nih.gov/40742806/ |
| Stress granules | Peri-implantitis (PI) | Immunofluorescence staining of G3BP and HuR proteins in gingival tissues show stress granules (SGs) aggregation in peri-implantitis (PI) gingival tissues but rarely in normal gingival tissues. Western blot analysis of healthy human gingival fibroblasts (hGFs) stimulated with P. gingivalis-lipopolysaccharide (LPS) or hypoxic conditions via 1% O2 stimulation show upregulated expression of HuR and G3BP, and the phosphorylation level of IRE1. Additionally, LPS or 1% O2 stimulation increased eIF2α phosphorylation and decreased 4EBP1 phosphorylation, with the combination more significant. Immunofluorescence reveals LPS or 1% O2 treatment induced SG formation, with the combination being more significant. In hGFs stimulated by LPS and 1% O2 co-treatment, PKR or 4EBP1 knockdown inhibited SGs aggregation and enhanced TNF-α and IL-1β expression. | https://pubmed.ncbi.nlm.nih.gov/40787261/ |
| Stress granules | Non-obstructive azoospermia (NOA) | Testicular biopsy samples from non-obstructive azoospermia (NOA) patients, but not testicular torsion controls exhibit robust phospho-MLKL and stress granule (SG) markers in all cases examined. In L929 mouse fibroblast cells, environmental stress triggers eIF2α kinase activation and SG assembly, which recruits ZBP1 and RIPK3 to initiate necroptosis. This assembly triggers RIPK3 activation, MLKL phosphorylation, and necroptotic death of mouse spermatogonia (PIWIL4) and Sertoli (SOX9) cells. Genetic ablation of Zbp1 or Ripk3 protects mice from heat-induced testicular degeneration, and activation of this pathway is also observed in aged human testes. | https://pubmed.ncbi.nlm.nih.gov/40811463/ |
| Stress granules | Drug resistance|Gastric cancer | UBAP2L expression in primary tumor samples from gastric cancer (GC) patients who underwent oxaliplatin-based chemotherapy indicate that UBAP2L upregulation is associated with poor chemotherapy responses. Knockdown of UBAP2L had no impact on cell proliferation under normal growth conditions, but impaired resistance of different GC cell lines to oxaliplatin and impeded the migration and invasion abilities of GC cell lines. Oxaliplatin-sensitive GC cells (parental cells) with or without UBAP2L overexpression implanted into nude mice show that UBAP2L overexpression results in tumor progression compared to controls under oxaliplatin treatment, and vehicle-treated tumors showed no significant difference between groups. Additionally, UBAP2L overexpression led to increased stress granule (SG) formation, along with a significant decrease in CC3-positive or TUNEL-positive area under oxaliplatin treatment. Co-immunoprecipitation (CoIP) and immunofluorescence (IF) assays in SGC-7901 cells show that in response to oxaliplatin, G3BP1 and RACK1, but not TIA1, co-immunoprecipitated with UBAP2L and was recruited to SGs. Furthermore IP followed by western blotting reveal transcriptional upregulation of UBAP2L is enhanced by oxaliplatin-induced phosphorylation and activation of HSF1 via AKT. Inhibiting either SG via ISRIB or HSF1 via DTHIB significantly overcomes oxaliplatin resistance in TDOXR mouse models, with the combination of oxaliplatin with ISRIB or DTHIB resulting in reduced levels of SGs and increased CC3-positive or TUNEL-positive areas. | https://pubmed.ncbi.nlm.nih.gov/40804300/ |
| Galectin-3-integrin α5β1 condensates | Diabetic wound healing | Immunohistochemical staining of Gal-3 in human diabetic (DM) and non-diabetic (Non-DM) skin sections show Gal-3 promotes diabetic wound healing and angiogenesis by binding to integrin α5β1 and potentiating its activation. In insulin-resistant HMEC-1 cells, cell migration and tube formation assays show Gal-3 promotes angiogenesis by binding to integrin α5β1 and potentiating its activation. Additionally, blockage of the integrin α5β1-FAK pathway inhibits the beneficial effect of Gal-3 on diabetic wound healing in streptozotocin (STZ)-induced diabetic rats. Confocal and fluorescence microscopy of HMEC-1 cells reveals Gal-3 binds to integrin α5β1 to induce liquid-liquid phase separation (LLPS) that promotes angiogenesis. Additionally, in vitro FRAP of Gal-3/integrin β1, Gal-3/integrin α5β1, and Gal-3/CD146 condensates recovered in ~90 s, ~60 s and ~80 s, respectively. In CHO-K1 cells, FRAP experiments illustrated a rapid recovery of the fluorescence of Gal-3/integrin α5 condensates in cells after photobleaching, and phosphorylation of FAK induced by Gal-3 was entirely blocked by lactose or 1,6-HD, highlighting the critical role of phase separation in Gal-3-mediated integrin α5β1 downstream activation. Upon addition of GFP-Gal-3 to cells, Gal-3, integrin α5 and CD146 formed co-phase separated droplets on the cell membrane and were disrupted by lactose treatment but not sucrose. Topical treatment of recombinant galectin-3 in hydrogels promoted diabetic wound healing in STZ-induced diabetic rats without causing systemic insulin resistance and synergizes with insulin. | https://pubmed.ncbi.nlm.nih.gov/40775187/ |
| Postsynaptic density | Phelan-McDermid syndrome | Reconstituted postsynaptic densities (PSDs) condensates form a soft glass material without signs of irreversible amyloid structure formation. PSD condensate formation is based on percolation of the PSD protein network via specific and multivalent interactions among scaffold proteins. Disruption of Shank3 SAM domain-mediated oligomerization, one type of SHANK3 mutation observed in Phelan-McDermid syndrome patients, softened the PSD condensate by weakening its network percolation, impaired synaptic transmission and plasticity, and caused autistic-like behavior in mice. | https://pubmed.ncbi.nlm.nih.gov/40848728/ |
| Stress granules | Osteosarcoma (OS) | High levels of AEP and truncated G3BP1 were found in osteosarcoma (OS) and glioma patient tumor tissues, and were strongly associated with poor prognosis. In an OS mouse model, inhibition of AEP through RR-11a promoted the tumor-suppressing effect of cisplatin, whereas AEP-cleaved G3BP1 rescue reversed the effect in both OS and glioma models. Chemotherapeutic drugs cisplatin, doxorubicin, etoposide, and methotrexate induced SG assembly and triggered AEP to specifically cleave G3BP1 at N258/N309 in U2OS, 143B, U87-MG and A549 cells. tG3BP1-Ns modulated SG dynamics by competitively binding to full-length G3BP1. In HeLa cells, tG3BP1-Cs sequestered mRNAs of ribosomal proteins and oxidative phosphorylation genes in the nucleoli and mitochondria to repress translation and oxidative stress. | https://pubmed.ncbi.nlm.nih.gov/40858563/ |
| SRSF9 condensates | Ovarian cancer | SRSF9 formed tumor-promoting phase-separated condensates within the nucleus, which were also important in splicing function. Specifically, NUMB mRNA m6A modification antagonized SRSF9-mediated alternative splicing. Additionally, loss of SRSF9 or antisense oligonucleotides-mediated isoform switch of NUMB mRNA inhibited OC growth in cells. SRSF9 was aberrantly upregulated in ovarian cancer (OC), correlating with poor patient prognosis. Loss of SRSF9 or antisense oligonucleotides-mediated isoform switch of NUMB mRNA inhibited OC growth in vivo. | https://pubmed.ncbi.nlm.nih.gov/40773673/ |
| RPS27L condensates | Myogenesis | Muscle-specific Rps27l knock-in (M─KI) mice exhibit significantly increased muscle mass, enlarged myofiber size, a higher proportion of fast-twitch myofibers, and enhanced muscle regeneration capabilities compared to WT controls. Overexpression of Rps27l promotes myoblast proliferation while inhibiting differentiation in skeletal muscle cells. Additionally, expression of Rps27l is negatively regulated by SIX4. The N-terminal intrinsically disordered region of RPS27L facilitates liquid-liquid phase separation (LLPS) and interacts with IGF1 to collaboratively regulate myogenesis. Additionally, droplets of recombinant GFP-RPS27L were observed in vitro via confocal microscopy, and exhibited concentration and salt-dependance. | https://pubmed.ncbi.nlm.nih.gov/40886325/ |
| EML4-ALK condensates | Cancer | Immunofluorescence and live cell imaging of H3122 cells show that EML4-ALK spontaneously forms biomolecular condensates. Furthermore, comparison of localization patterns in BEAS-2B cells reveals that EML4-ALK full-length (FL) forms cytoplasmic condensates, while a EML4-ALK ΔTD mutant and a EML4-ALK ΔHELP mutant exhibited a diffuse cellular distribution with no observable condensates. Dissolution of EML4-ALK condensates occurred in the presence of a TD-interfering peptide (TIP) in H3122 cells. Immunoblotting demonstrates a drastic decrease in the phosphorylated form of EML4-ALK (pEML4-ALK) in TIP-expressing cells, cell viability assays showed that TIP-expressing cells exhibited significantly slower growth, and colony forming assays show TIP-expressing cells produced significantly fewer colonies. Furthermore, TIP expression in EML4-ALK-negative cell lines (human lung epithelial BEAS-2B cells and non-small lung cancer A549 cells) had no effect on cell proliferation or clonogenicity. | https://pubmed.ncbi.nlm.nih.gov/40875402/ |
| Stress granules | Amyotrophic lateral sclerosis (ALS) | Immunocytochemistry shows that stress granule (SG) component G3BP1 exhibits enhanced interaction with Nup107 in motor neurons derived from patient iPSCs carrying C9orf72 Amyotrophic lateral sclerosis (ALS) mutations, where C9-1 and C9-2 mutant lines exhibited an increased number of SGs and slower SG disassembly following arsenite treatment compared to controls. Co-immunoprecipitation of G3BP1 in iPSC-derived motor neurons (iMNs) revealed increased interactions of G3BP1 with 40 SG components. Neurodegeneration-relevant proteins HSPH1, UBA1, SEP9, and DDX6 were among detected proteins, and immunostaining experiments confirm DDX6 foci colocalize with G3BP1 in SGs. Moreover, Nup107 colocalizes with SGs and aggregates in C9-ALS iMNs. Knockdown of npp-5, the C. elegans ortholog of Nup107, alleviates ALS-associated phenotypes in worm models, including dipeptide repeats production via repeat-associated non-AUG translation, progressive motility decline, and reduced lifespan. Knockdown of npp-5 ameliorated the short lifespan phenotype of C9ubi ALS worms, whereas knockdown of a different nucleoporin, npp-10 (Nup98 ortholog), did not extend the lifespan of C9ubi ALS worms. | https://pubmed.ncbi.nlm.nih.gov/40891053/ |
| ALYREF condensates | Cardiotoxicity|Drug-induced cardiotoxicity | Western blot analysis reveals that ALYREF expression was significantly reduced in doxorubicin (DOX)-treated mouse hearts, mouse cardiomyocytes (CMs), and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). In cardiac-specific ALYREF knockout mice (ALYREF CKO), echocardiography revealed that ALYREF CKO mice exhibited significant cardiac dysfunction, elevated creatine kinase-MB (CK-MB) and cardiac troponin T (cTnT), and enlarged left ventricular chambers and pathological cardiac hypertrophy. Cardiac-specific overexpression of ALYREF attenuates DOX-induced cardiotoxic injury in mice; following DOX treatment, mice with ALYREF overexpression exhibited improved survival and preserved body weight compared with controls. Echocardiographic analysis demonstrated that cardiac-specific overexpression of ALYREF significantly preserved left ventricular function, lowered serum levels of CK-MB and cTnT, and mitigation of DOX-induced myocardial atrophy and fibrosis compared to controls. Immunostaining in CMs show that following DOX exposure, ALYREF lost punctate nuclear organization and became diffusely distributed throughout the nucleoplasm with no nuclear translocation. Live-cell imaging of CMs transfected with EGFP-ALYREF confirmed that EGFP-ALYREF formed discrete nuclear condensates, with FRAP recovery occurring within 5 minutes of photobleaching. DOX induced DNA damage by binding to D171 and disrupted ALYREF condensates, indicated by increased γH2AX foci, greater comet tail DNA percentage, and higher nuclear fragmentation. DOX treatment of CMs additionally resulted in decreased binding of ALYREF to RBMX, TOP1, and CDC5L, and mutation of the D171 residue disrupted NARC1 complex assembly, resulting in DNA damage and apoptosis. | https://pubmed.ncbi.nlm.nih.gov/40899520/ |
| Stress granules | Acute lung injury (ALI) | Huashi baidu granules (HBG) decreased lung inflammation, mitigated fibrotic progression, and achieved near-complete structural restoration in a lipopolysaccharide (LPS)-induced acute lung injury (ALI) rat model. Furthermore, protein chip analysis revealed LPS-induced upregulation of pro-inflammatory cytokines IL-1β, IL-2, and TNF-α in rat lung tissues which were significantly attenuated by HBG treatment. Cell viability assays showed that HBG promoted A549 alveolar epithelial cell proliferation without cytotoxicity. In THP-1-derived M1-Mφs macrophages, ELISA analysis revealed that HBG suppressed LPS-induced IL-1β and TNF-α secretion by 72.1% and 63.1%, respectively. Flow cytometry analysis demonstrated that A549 cells exposed to conditioned media from treated M1-Mφs macrophages for 24 hours had reduced early apoptosis. RT-qPCR and western blot analyses revealed LPS-induced upregulation of G3BP1 and TIA1 mRNA/protein, with HBG normalizing expression levels. Immunofluorescence colocalization analysis confirmed elevated G3BP1/TIA1 fluorescence and SGs formation in an LPS-induced ALI rat model. HBG restored SGs colocalization rates to near-physiological levels. | https://pubmed.ncbi.nlm.nih.gov/40930288/ |
| Perineuronal nets | The behavior of male mice assigned to group-housed, isolated, or isolated/regrouped conditions from weaning age until end of adolescence (postnatal day 21 to 67; N = 8-10/group) were assessed in the open field, novel object recognition, Y-maze, and social interaction tests. Quantification of the number of parvalbumin-expressing (PV) cells and perineuronal nets (PNNs) were quantified in the prefrontal cortex (PFC), dorsal and ventral hippocampus. Results showed that early isolation led to hypersociability, which was rescued by regrouping; however regrouping also increased anxiety. Isolation reduced the proportion of PV cells surrounded by PNNs in the PFC, and regrouping reversed this effect and increased the total PNN count in PFC while it decreased PV/PNN count in the ventral hippocampus. | The behavior of male mice assigned to group-housed, isolated, or isolated/regrouped conditions from weaning age until end of adolescence (postnatal day 21 to 67; N = 8-10/group) were assessed in the open field, novel object recognition, Y-maze, and social interaction tests. Quantification of the number of parvalbumin-expressing (PV) cells and perineuronal nets (PNNs) were quantified in the prefrontal cortex (PFC), dorsal and ventral hippocampus. Results showed that early isolation led to hypersociability, which was rescued by regrouping; however regrouping also increased anxiety. Isolation reduced the proportion of PV cells surrounded by PNNs in the PFC, and regrouping reversed this effect and increased the total PNN count in PFC while it decreased PV/PNN count in the ventral hippocampus. | https://pubmed.ncbi.nlm.nih.gov/40975513/ |
| Nucleolus | Acute myeloid leukaemia (AML) | Growth competition assays in AML cell lines reveal that FBL knockdown resulted in increased cell cycle arrest (G0/G1 phase arrest), myeloid differentiation (CD11b+) and cell death, while flow cytometry show diminished leukaemia stem and progenitor cells (LSPCs; Lin−CD34+CD38−) and leukaemia stem cells (LSCs; Lin−CD34+CD38−CD99+Tim3+) after FBL depletion. Furthermore, primary AML patient cells confirm growth defects, myeloid differentiation, increased apoptosis and diminished colony formation upon FBL silencing. In immunodeficient non-obese diabetic (NOD)/severe combined immunodeficiency (SCID) mice transplanted with AML cells, knockdown of FBL led to slower leukaemia progression and extended survival, few leukaemic blast cells in the bone marrow, spleen, and liver, and a significant reduction in leukaemia cell frequencies in peripheral blood, bone marrow, and spleen. In HL-60 cells, deletion of the intrinsically disordered GAR domain and a chimeric FBL mutant, H2B-MD, with the GAR domain substituted with a non-IDR sequence from H2B failed to recover AML characteristics. Full restoration was observed using the MD-GAR mutant containing GAR and MD in revised order and the chimeric RGG-MD mutant with the FBL IDR replaced with a similar glycine- and arginine-rich sequence from NOLA1. Live-cell imaging of AML cells show GAR-defective mutants (MD, H2B-MD and PLD-MD) generated smaller puncta, failing to fully separate from fibrillar centre proteins and forming an incomplete hollow structure while RBD-defective mutants (GAR and GAR-Helix) exhibited fewer and larger condensates that dispersed across the nucleus yet still excluded fibrillar centre proteins. FRAP revealed GAR-defective mutants displayed higher mobility, as evidenced by faster fluorescence recovery after bleaching, and RBD-defective mutants (GAR and GAR-Helix) exhibited limited dynamics. | https://pubmed.ncbi.nlm.nih.gov/38745030/ |
| Stress granules | Clear renal cell carcinoma (ccRCC) | Patient-derived xenograft (PDX) and patient-derived organoid (PDO) models from samples obtained from a surgically resected clear renal cell carcinoma (ccRCC) patient treated with sunitinib examined by Western blot analysis show increased expression of UBAP2L. Additionally, in RCC cell lines 786-O and Caki-1, sunitinib-resistant cell lines exhibited increased UBAP2L protein expression as compared to parenteral cell lines. ccRCC patients with high UNAP2L expression also showed significantly higher resistance to sunitinib as compared with patients with ccRCC with low UBAP2L expression. In RCC cells lines, UBAP2L knockdown resulted in diminished cell viability, decreased maximal inhibitory concentration (IC50) values, and a reduction in both the number and size of colonies in ccRCC cells following sunitinib treatment, whereas overexpression elicited an opposing response. Furthermore, UBAP2L knockdown significantly increased the sensitivity of 786-O-R and Caki-1-R cells to sunitinib and promoted apoptosis, while overexpression decreased apoptosis and contributed to sunitinib resistance. In orthotopic xenograft model mice, sunitinib combined with UBAP2L knockdown impressively mitigated tumor growth compared with the control group, whereas UBAP2L knockdown or sunitinib alone exhibited a partial inhibitory effect. UBAP2L colocalized with stress granule (SG) marker protein G3BP1 following sunitinib treatment, and knockdown of UBAP2L disrupted SG organization--following sunitinib treatment, UBAP2L knockdown cells exhibited significantly lower levels of G3BP1 accumulation in the SG components. Additionally, immunoprecipitation and immunoblot found that S305 is the principal O-GlcNAcylation site of UBAP2L. UBAP2L S305A mutants exhibited reduced cell viability, diminished colony size and number, reduced angiogenic capacity, and disrupted SG formation in ccRCC cells. UBAP2L S305A mutant group showed slower tumor growth, smaller tumor size, and lower tumor weight in mice. Manipulation of O-GlcNAcylation via OSMI-1 treatment in tandem with sunitinib treatment shows inhibited cell growth and disrupted SG formation in ccRCC cells, slower tumor growth and enhanced survival in mouse models, and significant growth inhibition in PDOs. | https://pubmed.ncbi.nlm.nih.gov/41029457/ |
| Plectin condensates | Osteoporosis | Plectin (Plec) enhances osteoblast differentiation and bone formation via its intrinsically disordered region (IDR)-mediated phase separation. Plectin promotes osteoblast differentiation by sequestering Anxa2 through phase separation. Furthermore, the functional amino acid region (aa 1967–2185) of plectin exhibited therapeutic effect in ovariectomized and hindlimb unloading (HLU) mice. | https://pubmed.ncbi.nlm.nih.gov/41047030/ |
| Stress granules | Glioblastoma (GBM) | Lobeline impairs glioblastoma (GBM) cell survival by impairing SG disassembly after hypoxia in immortalized U251 and primary human U3085 GBM cells. After a hypoxic challenge, lobeline "locked" cells in a stressed state, even after re-exposure to normoxia. This is characterized by retained SGs, elevated levels of phosphorylated eIF2α and a sustained reduction in global protein translation. The disruption of the canonical stress response induced by lobeline resulted in increased cell death in both GBM cell lines. | https://pubmed.ncbi.nlm.nih.gov/41053069/ |
| α-synuclein condensates | Polyhalogenated carbazoles neuronal toxicity | Polyhalogenated carbazoles neuronal toxicity | https://pubmed.ncbi.nlm.nih.gov/41064882/ |
| CEBPZ condensates | Liver fibrosis | Baicalein (BA) significantly attenuated liver injury (AST and AST decreasing), inflammation (interleukin-6, interleukin-8 and tumor necrosis factor-alpha decreasing, and collagen deposition in carbon tetrachloride (CCl4)-induced fibrotic mice. In transforming growth factor-β1-activated LX-2 human hepatic stellate cells (HSC), BA suppressed cellular activation while inducing senescence through G1-phase cell cycle arrest and glycolysis inhibition (lactate decreasing). BA directly bound to CEBPZ, suppressing its LLPS and subsequently enhancing TP53 transcription. This activation of the p53/HK2 axis drove HSCs senescence and attenuated fibrosis. Pharmacological inhibition of p53 with pifithrin-α hydrobromide abolished BA's anti-fibrotic effects. BA's anti-fibrotic efficacy was entirely lost in CEBPZ-knockdown models. | https://pubmed.ncbi.nlm.nih.gov/41075517/ |
| Paraspeckles | Metabolic dysfunction-associated steatohepatitis (MASH)|Hepatocellular carcinoma (HCC) | Palmitic acid (PA) and oleic acid (OA) applied to tumor-tissue derived organoids and hepatocellular carcinoma (HCC) cell lines (HepG2, HCCLM3, Huh7, Hep3B) were employed to imitate pathological lipid overload in metabolic dysfunction-associated steatohepatitis (MASH). Organoids reveal m6A-modified circTACC3 is up-regulated in MASH-related HCC. Additionally, circTACC3 functionally facilitates the growth, lipid accumulation, and adaptive survival of organoids and HCC cells under lipid overload condition in a manner depending on its expression and location. circTACC3 interacted directly with NONO/p54nrb to assemble intranuclear paraspeckles. Fluorescence lifetime imaging microscopy-Forster resonance energy transfer (FLIM-FRET) assays further validated a direct interaction between circTACC3 and NONO/p54nrb. This process was dependent on the m6A-modification sites of circTACC3 and facilitated its nuclear retention. DRIP-ChIRP-sequencing demonstrated that circTACC3-containing paraspeckles were recruited to DNA double-strand break (DSB) foci to form R loops (DSB-circTACC3-R loops). DSB-circTACC3-R loops further facilitated the contact and fusion of topologically associated domains (TADs) and selectively activated genes related to the malignant phenotype of MASH-related HCC. circTACC3-R loops exerted positive feedback control over the assembly of circTACC3 paraspeckle and clustering of TADs. | https://pubmed.ncbi.nlm.nih.gov/41103024/ |
| α-synuclein condensates | Chronic intermittent hypoxia (CIH)-induced neuron damage | In a chronic intermittent hypoxia (CIH) mouse model (exposed to intermittent hypoxia for 10 weeks), contextual memory tests and cued memory tests show that CIH exposure resulted in a significant impairment of fear memory. Immunohistochemical analysis showed a decrease in TH-positive neurons in the ventral tegmental area (VTA), and transmission electron microscopy (TEM) revealed cytoplasmic vacuolization, severe nuclear deformation, increased lipofuscin accumulation, and mitochondrial damage in CIH-exposed mice. Golgi staining also showed a decreased dendritic spine number, reduced to about half, and lower density in the CIH group. Immunofluorescence and western blot analysis confirmed decreased TH expression and vital synaptic proteins PSD-95 and SYN, along with increased α-syn expression in the VTA. Furthermore, microelectrode Array (MEA) showed that SH-SY5Y cells exposed to IH had significantly reduced action potential number, frequency, and amplitude. Additionally, immunofluorescence staining revealed that within the IH group, a marked reduction in fluorescence intensity of TH, PSD-95, and SYN occurred, while fluorescence of α-syn was enhanced. Interfering with α-syn expression by siSNCA alleviated the decrease in electrophysiological activity, improved function, and had higher levels of TH, PSD-95, and SYN, with increases of approximately 2-fold, 2-fold, and 10-fold, respectively in SH-SY5Y cells. FRAP experiments show that IH induced abnormal phase separation of α-syn, leading to solid-like phase transitions and aggregation, which impeded fluorescence recovery. Confocal microscopy revealed that, with prolonged incubation times, the number and volume of α-syn droplets increased following H₂O₂ treatment in SH-SY5Y cells. DCFH-DA fluorescent probe was utilized to reveal that under IH conditions in SH-SY5Y cells and CIH exposure in mice, ROS fluorescence intensity increased in SH-SY5Y cells and ROS content in the CIH group was approximately 3-fold higher, respectively. In SH-SY5Y cells, treatment with Danshensu methyl ester (DME) reduced ROS levels and inhibited α-syn abnormal phase separation. FRAP experiments demonstrated that the fluorescence intensity of α-syn droplets in SH-SY5Y cells recovered to approximately 80% of the initial value within 5 min post-bleaching when treated with DME, indicating high fluidity of these droplets. In mice, treatment with DME increased neuronal action potential number, frequency, and amplitude. Additionally, western blot and Immunofluorescence staining showed α-syn aggregation was decreased, while expression of TH, PSD-95, and SYN proteins was increased. DME also alleviated the dopaminergic neuron damage and fear memory impairment caused by CIH. | https://pubmed.ncbi.nlm.nih.gov/41162854/ |