{"gene":"VIL1","run_date":"2026-06-14T21:23:28+00:00","timeline":{"discoveries":[{"year":2017,"finding":"VIL1 (villin-1) and gelsolin (GSN) are required for dephosphorylation of EIF2A (eukaryotic translation initiation factor 2 subunit alpha) and recovery from cell stress in intestinal epithelial cells (IECs). Under acute stress, EIF2A signaling reduces VIL1 and GSN expression; during prolonged stress, continued downregulation of VIL1 and GSN leads to constitutive EIF2A phosphorylation, IRGM1 overexpression, and necroptotic cell death. VIL1/GSN double-knockout mice develop spontaneous ileitis resembling Crohn's disease.","method":"Villin-1/gelsolin double-knockout mice, histology, immunoblots, phalloidin staining, immunohistochemistry, electron microscopy, flow cytometry, lentiviral GFP-EIF2A constructs, human CD patient tissue analysis","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO mouse model with defined cellular phenotype, multiple orthogonal methods (IHC, immunoblots, flow cytometry, electron microscopy), corroborated in human CD tissue","pmids":["29274870"],"is_preprint":false},{"year":2024,"finding":"VIL1 knockout in colorectal cancer (CRC) cells activates ferroptosis and inhibits migration, while VIL1 overexpression inhibits ferroptosis and promotes tumor growth. Mechanistically, VIL1 binds to NF-κB p105 and controls NF-κB expression; in vivo, VIL1 overexpression induces NF-κB and lipocalin-2 (LCN2) expression, identifying the VIL1/NF-κB axis as a regulator of CRC progression through ferroptosis modulation.","method":"VIL1 knockout and overexpression in CRC cell lines, in vivo tumor xenograft, transcriptomics, immunoblotting, cell proliferation and migration assays, apoptosis assays, ferroptosis assays","journal":"The FEBS journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO/OE with defined phenotypic readouts and mechanistic pathway placement (NF-κB binding), single lab, multiple orthogonal methods","pmids":["39658274"],"is_preprint":false},{"year":2025,"finding":"The enterotoxin Euphorin G from Euphorbia fischeriana covalently modifies Cys624 of Villin-1, causing F-actin disassembly and intestinal barrier failure. Chebulinic acid from Terminalia chebula protects Villin-1 by non-covalently binding it and shielding Cys624 from covalent attack, while allosterically enhancing Villin-1's F-actin binding affinity, thereby preserving intestinal barrier integrity.","method":"Intestinal organoid models, chemical biology, proteomics, Villin-1 knockout and Cys624 mutagenesis, in vitro F-actin binding assays, in vivo validation of barrier integrity","journal":"Journal of ethnopharmacology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — site-directed mutagenesis of active-site cysteine (Cys624), in vitro reconstitution of F-actin binding, genetic KO validation, multiple orthogonal methods in one study","pmids":["40835194"],"is_preprint":false},{"year":2014,"finding":"In euryhaline medaka fish, the villin-1 homolog VILL is localized to the apical region of gill ionocytes and is required for formation of apical microvilli. VILL expression is induced by hypoosmotic conditions (freshwater acclimation), and morpholino knockdown of VILL eliminates apical protrusions of ionocytes and pavement cells, establishing a direct role for this villin-1 ortholog in actin-based microvillus formation in absorptive cells.","method":"Immunofluorescence localization, morpholino knockdown, scanning electron microscopy, quantitative Western blot and RT-PCR across salinity conditions","journal":"Frontiers in zoology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — morpholino loss-of-function with defined structural phenotype, immunofluorescence localization, multiple methods; ortholog of mammalian VIL1 in a fish model, single lab","pmids":["24410933"],"is_preprint":false},{"year":2017,"finding":"Plasmatic villin-1 levels correspond with the severity of kidney injury during renal ischemia-reperfusion, and its release into plasma is associated with redistribution of villin-1 from proximal tubular brush-border cells. Treatment with necrostatin-1 (a necroptosis inhibitor) decreased plasmatic villin-1 levels earlier than other markers, linking villin-1 release to necroptotic cell death in proximal tubular cells.","method":"Immunohistochemistry on kidney sections, Western blotting of plasma from rat ischemia-reperfusion model and pig renal transplantation model, and in liver transplant patients developing AKI; necrostatin-1 pharmacological intervention","journal":"Transplantation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — immunohistochemistry plus Western blot across multiple species and models, pharmacological intervention linking necroptosis to VIL1 release, single lab","pmids":["28704336"],"is_preprint":false},{"year":2026,"finding":"In villin-1/gelsolin double-knockout mice, chronic integrated stress response (ISR) activation and RIPK3-mediated necroptosis converge to drive epithelial injury, Paneth cell expansion, and barrier dysfunction. Pharmacologic inhibition of ISR or RIPK3 (using ISR inhibitor, necrostatin-1, pazopanib, or ponatinib) restored villus architecture, epithelial survival, regeneration, and transepithelial electrical resistance, placing VIL1 loss upstream of ISR/necroptosis-mediated epithelial dysfunction.","method":"Villin-1/gelsolin double-knockout mice, Tnf mice, CD patient-derived enteroids, ISR inhibitor and RIPK3 inhibitor treatments, transepithelial electrical resistance measurement, enteroid formation/budding assays","journal":"Gastro hep advances","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO mouse and patient-derived organoid models, pharmacological rescue, multiple functional readouts; single lab extending prior findings","pmids":["42254860"],"is_preprint":false}],"current_model":"VIL1 (villin-1) is an actin-regulatory protein of intestinal epithelial brush borders whose Cys624 residue directly mediates F-actin binding and is subject to covalent modification; in intestinal epithelial cells it is required for dephosphorylation of EIF2A and recovery from cell stress, such that its chronic loss leads to constitutive ISR activation, RIPK3-mediated necroptosis, and intestinal inflammation, while in colorectal cancer it suppresses ferroptosis by binding NF-κB and driving LCN2 expression, and its release into plasma or urine serves as a marker of proximal tubular or gut epithelial barrier injury."},"narrative":{"mechanistic_narrative":"VIL1 (villin-1) is an actin-regulatory protein of absorptive epithelial brush borders that organizes the apical actin cytoskeleton and maintains epithelial barrier integrity [PMID:29274870, PMID:40835194, PMID:24410933]. Its F-actin binding is mediated through Cys624: covalent modification of this residue by an enterotoxin triggers F-actin disassembly and barrier failure, while a protective small molecule that shields Cys624 allosterically enhances F-actin binding affinity and preserves barrier integrity [PMID:40835194]. Across absorptive cell types VIL1 (and its fish ortholog VILL) is required for formation of apical microvilli [PMID:24410933]. In intestinal epithelial cells, VIL1 together with gelsolin is required for dephosphorylation of EIF2A and recovery from cell stress; their chronic loss produces constitutive EIF2A phosphorylation and integrated stress response activation, IRGM1 overexpression, and RIPK3-mediated necroptotic death, with double-knockout mice developing spontaneous Crohn's-like ileitis that is reversed by ISR or RIPK3 inhibition [PMID:29274870, PMID:42254860]. In colorectal cancer cells VIL1 suppresses ferroptosis and promotes tumor growth by binding NF-κB p105 and driving NF-κB and lipocalin-2 (LCN2) expression [PMID:39658274]. Release of VIL1 from injured brush-border cells into plasma tracks the severity of necroptotic proximal tubular injury [PMID:28704336].","teleology":[{"year":2014,"claim":"Establishing whether villin-1 actively builds apical microvilli rather than merely decorating them, loss-of-function in a fish ortholog tied VILL directly to actin-based microvillus formation in absorptive cells.","evidence":"Morpholino knockdown of VILL in medaka gill ionocytes with immunofluorescence and scanning electron microscopy across salinity conditions","pmids":["24410933"],"confidence":"Medium","gaps":["Ortholog in a fish model, not direct human VIL1","Does not define the molecular mechanism by which VILL nucleates or bundles apical actin"]},{"year":2017,"claim":"Connecting brush-border actin regulators to epithelial stress recovery, VIL1 and gelsolin were shown to be required for EIF2A dephosphorylation, linking their loss to constitutive stress signaling, necroptosis, and Crohn's-like inflammation.","evidence":"Villin-1/gelsolin double-knockout mice with histology, immunoblots, flow cytometry, electron microscopy, and corroboration in human CD tissue","pmids":["29274870"],"confidence":"High","gaps":["Molecular link between VIL1 and the EIF2A phosphatase machinery not resolved","Whether the effect requires VIL1's actin-binding activity is untested"]},{"year":2017,"claim":"Testing whether VIL1 release reports epithelial death, plasmatic villin-1 was shown to rise with proximal tubular injury and to depend on necroptosis, validating it as a necroptosis-linked injury marker.","evidence":"Immunohistochemistry and Western blotting across rat, pig, and human renal/liver injury models with necrostatin-1 intervention","pmids":["28704336"],"confidence":"Medium","gaps":["Mechanism of VIL1 redistribution and release not defined","Single lab; biomarker specificity versus other brush-border proteins unclear"]},{"year":2024,"claim":"Asking how VIL1 influences tumor cell fate, knockout/overexpression in colorectal cancer cells placed VIL1 upstream of ferroptosis suppression via direct NF-κB p105 binding and LCN2 induction.","evidence":"VIL1 knockout/overexpression in CRC lines with xenografts, transcriptomics, immunoblotting, and ferroptosis assays","pmids":["39658274"],"confidence":"Medium","gaps":["Direct VIL1–NF-κB binding interface not mapped","Relationship between VIL1's actin role and NF-κB regulation unknown","Single lab"]},{"year":2025,"claim":"Identifying the molecular site of VIL1's actin control, Cys624 was shown to be the covalently targetable residue governing F-actin binding and barrier integrity.","evidence":"Intestinal organoids, chemical proteomics, Cys624 mutagenesis, in vitro F-actin binding assays, and in vivo barrier validation","pmids":["40835194"],"confidence":"High","gaps":["Structural basis of Cys624-dependent actin binding not resolved","Whether physiological modification of Cys624 occurs endogenously is unknown"]},{"year":2026,"claim":"Resolving the causal hierarchy of VIL1-loss pathology, ISR and RIPK3 were placed downstream of VIL1 loss, with pharmacologic inhibition rescuing epithelial architecture and barrier function.","evidence":"Villin-1/gelsolin double-knockout mice and CD patient-derived enteroids with ISR/RIPK3 inhibitor rescue and transepithelial resistance readouts","pmids":["42254860"],"confidence":"Medium","gaps":["Direct biochemical step connecting VIL1 loss to ISR activation still undefined","Single lab extending prior findings"]},{"year":null,"claim":"How VIL1's actin-bundling activity at the brush border mechanistically couples to its roles in EIF2A dephosphorylation and NF-κB binding remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model linking actin binding to stress-signaling functions","Whether actin-independent scaffolding underlies the NF-κB and ISR roles is untested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2,3]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[2,3]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3,4]}],"pathway":[{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[0,5]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,5]}],"complexes":[],"partners":["GSN","ACTB","NFKB1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P09327","full_name":"Villin-1","aliases":[],"length_aa":827,"mass_kda":92.7,"function":"Epithelial cell-specific Ca(2+)-regulated actin-modifying protein that modulates the reorganization of microvillar actin filaments. Plays a role in the actin nucleation, actin filament bundle assembly, actin filament capping and severing. Binds phosphatidylinositol 4,5-bisphosphate (PIP2) and lysophosphatidic acid (LPA); binds LPA with higher affinity than PIP2. Binding to LPA increases its phosphorylation by SRC and inhibits all actin-modifying activities. Binding to PIP2 inhibits actin-capping and -severing activities but enhances actin-bundling activity. Regulates the intestinal epithelial cell morphology, cell invasion, cell migration and apoptosis. Protects against apoptosis induced by dextran sodium sulfate (DSS) in the gastrointestinal epithelium. Appears to regulate cell death by maintaining mitochondrial integrity. Enhances hepatocyte growth factor (HGF)-induced epithelial cell motility, chemotaxis and wound repair. Upon S.flexneri cell infection, its actin-severing activity enhances actin-based motility of the bacteria and plays a role during the dissemination","subcellular_location":"Cytoplasm, cytoskeleton; Cell projection, lamellipodium; Cell projection, ruffle; Cell projection, microvillus; Cell projection, filopodium tip; Cell projection, filopodium","url":"https://www.uniprot.org/uniprotkb/P09327/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/VIL1","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/VIL1","total_profiled":1310},"omim":[{"mim_id":"619666","title":"VILLIN-LIKE PROTEIN; VILL","url":"https://www.omim.org/entry/619666"},{"mim_id":"600266","title":"SOLUTE CARRIER FAMILY 11 (PROTON-COUPLED DIVALENT METAL ION TRANSPORTER), MEMBER 1; SLC11A1","url":"https://www.omim.org/entry/600266"},{"mim_id":"193040","title":"VILLIN 1; VIL1","url":"https://www.omim.org/entry/193040"},{"mim_id":"153615","title":"CAPPING PROTEIN, GELSOLIN-LIKE; CAPG","url":"https://www.omim.org/entry/153615"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Plasma membrane","reliability":"Enhanced"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"intestine","ntpm":410.7}],"url":"https://www.proteinatlas.org/search/VIL1"},"hgnc":{"alias_symbol":["D2S1471"],"prev_symbol":["VIL"]},"alphafold":{"accession":"P09327","domains":[{"cath_id":"3.40.20.10","chopping":"7-120","consensus_level":"high","plddt":82.1138,"start":7,"end":120},{"cath_id":"3.40.20.10","chopping":"132-240_720-750","consensus_level":"medium","plddt":75.3909,"start":132,"end":750},{"cath_id":"3.40.20.10","chopping":"249-346","consensus_level":"high","plddt":79.6218,"start":249,"end":346},{"cath_id":"3.40.20.10","chopping":"393-500","consensus_level":"high","plddt":85.3249,"start":393,"end":500},{"cath_id":"3.40.20.10","chopping":"517-607","consensus_level":"high","plddt":89.5501,"start":517,"end":607},{"cath_id":"3.40.20.10","chopping":"621-717","consensus_level":"high","plddt":77.8864,"start":621,"end":717},{"cath_id":"1.10.950.10","chopping":"770-825","consensus_level":"high","plddt":85.6795,"start":770,"end":825}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P09327","model_url":"https://alphafold.ebi.ac.uk/files/AF-P09327-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P09327-F1-predicted_aligned_error_v6.png","plddt_mean":77.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=VIL1","jax_strain_url":"https://www.jax.org/strain/search?query=VIL1"},"sequence":{"accession":"P09327","fasta_url":"https://rest.uniprot.org/uniprotkb/P09327.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P09327/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P09327"}},"corpus_meta":[{"pmid":"19956723","id":"PMC_19956723","title":"Mucosal gene expression of antimicrobial peptides in inflammatory bowel disease before and after first infliximab treatment.","date":"2009","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/19956723","citation_count":240,"is_preprint":false},{"pmid":"14655050","id":"PMC_14655050","title":"Down-regulation of a gastric transcription factor, Sox2, and ectopic expression of intestinal homeobox genes, Cdx1 and Cdx2: inverse correlation during progression from gastric/intestinal-mixed to complete intestinal metaplasia.","date":"2003","source":"Journal of cancer research and clinical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/14655050","citation_count":116,"is_preprint":false},{"pmid":"20616339","id":"PMC_20616339","title":"Tumor lymphangiogenesis and metastasis to lymph nodes induced by cancer cell expression of podoplanin.","date":"2010","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/20616339","citation_count":106,"is_preprint":false},{"pmid":"33494396","id":"PMC_33494396","title":"Proteomics Analysis of Gastric Cancer Patients with Diabetes Mellitus.","date":"2021","source":"Journal of clinical medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33494396","citation_count":62,"is_preprint":false},{"pmid":"26940098","id":"PMC_26940098","title":"Proteomics of Urinary Vesicles Links Plakins and Complement to Polycystic Kidney Disease.","date":"2016","source":"Journal of the American Society of Nephrology : JASN","url":"https://pubmed.ncbi.nlm.nih.gov/26940098","citation_count":62,"is_preprint":false},{"pmid":"29274870","id":"PMC_29274870","title":"Villin-1 and Gelsolin Regulate Changes in Actin Dynamics That Affect Cell Survival Signaling Pathways and Intestinal Inflammation.","date":"2017","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/29274870","citation_count":60,"is_preprint":false},{"pmid":"19899082","id":"PMC_19899082","title":"Differential expression proteomics of human colorectal cancer based on a syngeneic cellular model for the progression of adenoma to carcinoma.","date":"2010","source":"Proteomics","url":"https://pubmed.ncbi.nlm.nih.gov/19899082","citation_count":46,"is_preprint":false},{"pmid":"25605255","id":"PMC_25605255","title":"A three-protein signature and clinical outcome in esophageal squamous cell carcinoma.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/25605255","citation_count":41,"is_preprint":false},{"pmid":"19808654","id":"PMC_19808654","title":"Pdx1 inactivation restricted to the intestinal epithelium in mice alters duodenal gene expression in enterocytes and enteroendocrine cells.","date":"2009","source":"American journal of physiology. Gastrointestinal and liver physiology","url":"https://pubmed.ncbi.nlm.nih.gov/19808654","citation_count":40,"is_preprint":false},{"pmid":"19627535","id":"PMC_19627535","title":"High-grade neuroendocrine carcinoma of the lung: comparative clinicopathological study of large cell neuroendocrine carcinoma and small cell lung carcinoma.","date":"2009","source":"Pathology international","url":"https://pubmed.ncbi.nlm.nih.gov/19627535","citation_count":37,"is_preprint":false},{"pmid":"32350063","id":"PMC_32350063","title":"Regional Proteomic Quantification of Clinically Relevant Non-Cytochrome P450 Enzymes along the Human Small Intestine.","date":"2020","source":"Drug metabolism and disposition: the biological fate of chemicals","url":"https://pubmed.ncbi.nlm.nih.gov/32350063","citation_count":29,"is_preprint":false},{"pmid":"30894050","id":"PMC_30894050","title":"Atg14 protects the intestinal epithelium from TNF-triggered villus atrophy.","date":"2019","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/30894050","citation_count":28,"is_preprint":false},{"pmid":"15375555","id":"PMC_15375555","title":"Search for epithelial-specific mRNAs in peripheral blood of patients with colon cancer by RT-PCR.","date":"2004","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/15375555","citation_count":26,"is_preprint":false},{"pmid":"26398045","id":"PMC_26398045","title":"A quantitative proteomics study on olfactomedin 4 in the development of gastric cancer.","date":"2015","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/26398045","citation_count":25,"is_preprint":false},{"pmid":"21910169","id":"PMC_21910169","title":"Intestinal alpha-defensin expression in pediatric inflammatory bowel disease.","date":"2010","source":"Inflammatory bowel diseases","url":"https://pubmed.ncbi.nlm.nih.gov/21910169","citation_count":24,"is_preprint":false},{"pmid":"27013401","id":"PMC_27013401","title":"Decreased Pregnane X Receptor Expression in Children with Active Crohn's Disease.","date":"2016","source":"Drug metabolism and disposition: the biological fate of chemicals","url":"https://pubmed.ncbi.nlm.nih.gov/27013401","citation_count":21,"is_preprint":false},{"pmid":"31260507","id":"PMC_31260507","title":"Schlafen 3 knockout mice display gender-specific differences in weight gain, food efficiency, and expression of markers of intestinal epithelial differentiation, metabolism, and immune cell function.","date":"2019","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/31260507","citation_count":19,"is_preprint":false},{"pmid":"8020939","id":"PMC_8020939","title":"Regional mapping of loci from human chromosome 2q to sheep chromosome 2q.","date":"1994","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/8020939","citation_count":18,"is_preprint":false},{"pmid":"22530999","id":"PMC_22530999","title":"Villin 1 is a predictive factor for the recurrence of high serum alpha-fetoprotein-associated hepatocellular carcinoma after hepatectomy.","date":"2012","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/22530999","citation_count":17,"is_preprint":false},{"pmid":"37662988","id":"PMC_37662988","title":"Assessing the effects of a mixed Eimeria spp. challenge on performance, intestinal integrity, and the gut microbiome of broiler chickens.","date":"2023","source":"Frontiers in veterinary science","url":"https://pubmed.ncbi.nlm.nih.gov/37662988","citation_count":17,"is_preprint":false},{"pmid":"39377914","id":"PMC_39377914","title":"TTF-1 is a highly sensitive but not fully specific marker for pulmonary and thyroidal cancer: a tissue microarray study evaluating more than 17,000 tumors from 152 different tumor entities.","date":"2024","source":"Virchows Archiv : an international journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/39377914","citation_count":14,"is_preprint":false},{"pmid":"36161099","id":"PMC_36161099","title":"Effects of human induced pluripotent stem cell-derived intestinal organoids on colitis-model mice.","date":"2022","source":"Regenerative therapy","url":"https://pubmed.ncbi.nlm.nih.gov/36161099","citation_count":13,"is_preprint":false},{"pmid":"25059646","id":"PMC_25059646","title":"Inflammation-Induced Downregulation of Butyrate Uptake and Oxidation Is Not Caused by a Reduced Gene Expression.","date":"2015","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/25059646","citation_count":11,"is_preprint":false},{"pmid":"36683854","id":"PMC_36683854","title":"Poly(I:C)-exposed zebrafish shows autism-like behaviors which are ameliorated by fabp2 gene knockout.","date":"2023","source":"Frontiers in molecular neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/36683854","citation_count":10,"is_preprint":false},{"pmid":"28704336","id":"PMC_28704336","title":"Plasmatic Villin 1 Is a Novel In Vivo Marker of Proximal Tubular Cell Injury During Renal Ischemia-Reperfusion.","date":"2017","source":"Transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/28704336","citation_count":8,"is_preprint":false},{"pmid":"24410933","id":"PMC_24410933","title":"Medaka villin 1-like protein (VILL) is associated with the formation of microvilli induced by decreasing salinities in the absorptive ionocytes.","date":"2014","source":"Frontiers in zoology","url":"https://pubmed.ncbi.nlm.nih.gov/24410933","citation_count":8,"is_preprint":false},{"pmid":"37643848","id":"PMC_37643848","title":"Diagnostic potential of plasma biomarkers and exhaled volatile organic compounds in predicting the different stages of acute mesenteric ischaemia: protocol for a multicentre prospective observational study (TACTIC study).","date":"2023","source":"BMJ open","url":"https://pubmed.ncbi.nlm.nih.gov/37643848","citation_count":7,"is_preprint":false},{"pmid":"37731918","id":"PMC_37731918","title":"Lead induced structural and functional damage and microbiota dysbiosis in the intestine of crucian carp (Carassius auratus).","date":"2023","source":"Frontiers in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/37731918","citation_count":6,"is_preprint":false},{"pmid":"8747925","id":"PMC_8747925","title":"Thirteen loci physically assigned to sheep chromosome 2 by cell hybrid analysis and in situ hybridization.","date":"1995","source":"Mammalian genome : official journal of the International Mammalian Genome Society","url":"https://pubmed.ncbi.nlm.nih.gov/8747925","citation_count":5,"is_preprint":false},{"pmid":"34710177","id":"PMC_34710177","title":"Vil-Cre specific Schlafen 3 knockout mice exhibit sex-specific differences in intestinal differentiation markers and Schlafen family members expression levels.","date":"2021","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/34710177","citation_count":5,"is_preprint":false},{"pmid":"41340228","id":"PMC_41340228","title":"Serum Villin-1-A Novel Marker of Gut Barrier Damage in Acutely Decompensated Cirrhosis: A Cohort Study and Validation.","date":"2025","source":"Alimentary pharmacology & therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/41340228","citation_count":4,"is_preprint":false},{"pmid":"27557990","id":"PMC_27557990","title":"The expression of VILL protein is hypoosmotic-dependent in the lamellar gill ionocytes of Otocephala teleost fish, Chanos chanos.","date":"2016","source":"Comparative biochemistry and physiology. Part A, Molecular & integrative physiology","url":"https://pubmed.ncbi.nlm.nih.gov/27557990","citation_count":4,"is_preprint":false},{"pmid":"39658274","id":"PMC_39658274","title":"Villin-1 regulates ferroptosis in colorectal cancer progression.","date":"2024","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/39658274","citation_count":3,"is_preprint":false},{"pmid":"39584016","id":"PMC_39584016","title":"A customizable and low-cost 3D-printed transwell device coupled with 3D cell culture for permeability assay.","date":"2024","source":"HardwareX","url":"https://pubmed.ncbi.nlm.nih.gov/39584016","citation_count":3,"is_preprint":false},{"pmid":"41347923","id":"PMC_41347923","title":"Spasmolytic Polypeptide-Expressing Metaplasia (SPEM): the hidden linchpin in gastric carcinogenesis beyond the correa cascade.","date":"2025","source":"International journal of surgery (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/41347923","citation_count":2,"is_preprint":false},{"pmid":"38201306","id":"PMC_38201306","title":"An Approach to Intersectionally Target Mature Enteroendocrine Cells in the Small Intestine of Mice.","date":"2024","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/38201306","citation_count":1,"is_preprint":false},{"pmid":"39072182","id":"PMC_39072182","title":"Weiwei Decoction alleviates gastric intestinal metaplasia through the olfactomedin 4/nucleotide-binding oligomerization domain 1/caudal-type homeobox gene 2 signaling pathway.","date":"2024","source":"World journal of gastrointestinal oncology","url":"https://pubmed.ncbi.nlm.nih.gov/39072182","citation_count":1,"is_preprint":false},{"pmid":"40835194","id":"PMC_40835194","title":"Chebulinic acid shields Villin 1 from covalent attack to mitigate Euphorbia fischeriana enterotoxicity: A basis for safer anti-ascites therapy.","date":"2025","source":"Journal of ethnopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/40835194","citation_count":0,"is_preprint":false},{"pmid":"41772475","id":"PMC_41772475","title":"Comprehensive insights into the development of arabidopsis trichomes by single-nucleus RNA-seq.","date":"2026","source":"BMC plant biology","url":"https://pubmed.ncbi.nlm.nih.gov/41772475","citation_count":0,"is_preprint":false},{"pmid":"42254860","id":"PMC_42254860","title":"Integrated Stress Response and Necroptosis Drive Epithelial Dysfunction in Crohn's Disease: Repurposing Cancer Drugs for Permeability Barrier Healing.","date":"2026","source":"Gastro hep advances","url":"https://pubmed.ncbi.nlm.nih.gov/42254860","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"sonnet_model":"claude-sonnet-4-6","sonnet_input_tokens":26770,"sonnet_output_tokens":2032,"sonnet_usd":0.1108,"sonnet_elapsed_seconds":37.7,"stage1_stop_reason":"end_turn"},"stage2":{"opus_model":"claude-opus-4-8","opus_input_tokens":8961,"opus_output_tokens":2471,"opus_usd":0.1066,"opus_elapsed_seconds":27.2,"stage2_stop_reason":"end_turn"},"total_usd":0.2174},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2017,\n      \"finding\": \"VIL1 (villin-1) and gelsolin (GSN) are required for dephosphorylation of EIF2A (eukaryotic translation initiation factor 2 subunit alpha) and recovery from cell stress in intestinal epithelial cells (IECs). Under acute stress, EIF2A signaling reduces VIL1 and GSN expression; during prolonged stress, continued downregulation of VIL1 and GSN leads to constitutive EIF2A phosphorylation, IRGM1 overexpression, and necroptotic cell death. VIL1/GSN double-knockout mice develop spontaneous ileitis resembling Crohn's disease.\",\n      \"method\": \"Villin-1/gelsolin double-knockout mice, histology, immunoblots, phalloidin staining, immunohistochemistry, electron microscopy, flow cytometry, lentiviral GFP-EIF2A constructs, human CD patient tissue analysis\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO mouse model with defined cellular phenotype, multiple orthogonal methods (IHC, immunoblots, flow cytometry, electron microscopy), corroborated in human CD tissue\",\n      \"pmids\": [\"29274870\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"VIL1 knockout in colorectal cancer (CRC) cells activates ferroptosis and inhibits migration, while VIL1 overexpression inhibits ferroptosis and promotes tumor growth. Mechanistically, VIL1 binds to NF-κB p105 and controls NF-κB expression; in vivo, VIL1 overexpression induces NF-κB and lipocalin-2 (LCN2) expression, identifying the VIL1/NF-κB axis as a regulator of CRC progression through ferroptosis modulation.\",\n      \"method\": \"VIL1 knockout and overexpression in CRC cell lines, in vivo tumor xenograft, transcriptomics, immunoblotting, cell proliferation and migration assays, apoptosis assays, ferroptosis assays\",\n      \"journal\": \"The FEBS journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO/OE with defined phenotypic readouts and mechanistic pathway placement (NF-κB binding), single lab, multiple orthogonal methods\",\n      \"pmids\": [\"39658274\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The enterotoxin Euphorin G from Euphorbia fischeriana covalently modifies Cys624 of Villin-1, causing F-actin disassembly and intestinal barrier failure. Chebulinic acid from Terminalia chebula protects Villin-1 by non-covalently binding it and shielding Cys624 from covalent attack, while allosterically enhancing Villin-1's F-actin binding affinity, thereby preserving intestinal barrier integrity.\",\n      \"method\": \"Intestinal organoid models, chemical biology, proteomics, Villin-1 knockout and Cys624 mutagenesis, in vitro F-actin binding assays, in vivo validation of barrier integrity\",\n      \"journal\": \"Journal of ethnopharmacology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — site-directed mutagenesis of active-site cysteine (Cys624), in vitro reconstitution of F-actin binding, genetic KO validation, multiple orthogonal methods in one study\",\n      \"pmids\": [\"40835194\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In euryhaline medaka fish, the villin-1 homolog VILL is localized to the apical region of gill ionocytes and is required for formation of apical microvilli. VILL expression is induced by hypoosmotic conditions (freshwater acclimation), and morpholino knockdown of VILL eliminates apical protrusions of ionocytes and pavement cells, establishing a direct role for this villin-1 ortholog in actin-based microvillus formation in absorptive cells.\",\n      \"method\": \"Immunofluorescence localization, morpholino knockdown, scanning electron microscopy, quantitative Western blot and RT-PCR across salinity conditions\",\n      \"journal\": \"Frontiers in zoology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — morpholino loss-of-function with defined structural phenotype, immunofluorescence localization, multiple methods; ortholog of mammalian VIL1 in a fish model, single lab\",\n      \"pmids\": [\"24410933\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Plasmatic villin-1 levels correspond with the severity of kidney injury during renal ischemia-reperfusion, and its release into plasma is associated with redistribution of villin-1 from proximal tubular brush-border cells. Treatment with necrostatin-1 (a necroptosis inhibitor) decreased plasmatic villin-1 levels earlier than other markers, linking villin-1 release to necroptotic cell death in proximal tubular cells.\",\n      \"method\": \"Immunohistochemistry on kidney sections, Western blotting of plasma from rat ischemia-reperfusion model and pig renal transplantation model, and in liver transplant patients developing AKI; necrostatin-1 pharmacological intervention\",\n      \"journal\": \"Transplantation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — immunohistochemistry plus Western blot across multiple species and models, pharmacological intervention linking necroptosis to VIL1 release, single lab\",\n      \"pmids\": [\"28704336\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"In villin-1/gelsolin double-knockout mice, chronic integrated stress response (ISR) activation and RIPK3-mediated necroptosis converge to drive epithelial injury, Paneth cell expansion, and barrier dysfunction. Pharmacologic inhibition of ISR or RIPK3 (using ISR inhibitor, necrostatin-1, pazopanib, or ponatinib) restored villus architecture, epithelial survival, regeneration, and transepithelial electrical resistance, placing VIL1 loss upstream of ISR/necroptosis-mediated epithelial dysfunction.\",\n      \"method\": \"Villin-1/gelsolin double-knockout mice, Tnf mice, CD patient-derived enteroids, ISR inhibitor and RIPK3 inhibitor treatments, transepithelial electrical resistance measurement, enteroid formation/budding assays\",\n      \"journal\": \"Gastro hep advances\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO mouse and patient-derived organoid models, pharmacological rescue, multiple functional readouts; single lab extending prior findings\",\n      \"pmids\": [\"42254860\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"VIL1 (villin-1) is an actin-regulatory protein of intestinal epithelial brush borders whose Cys624 residue directly mediates F-actin binding and is subject to covalent modification; in intestinal epithelial cells it is required for dephosphorylation of EIF2A and recovery from cell stress, such that its chronic loss leads to constitutive ISR activation, RIPK3-mediated necroptosis, and intestinal inflammation, while in colorectal cancer it suppresses ferroptosis by binding NF-κB and driving LCN2 expression, and its release into plasma or urine serves as a marker of proximal tubular or gut epithelial barrier injury.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"VIL1 (villin-1) is an actin-regulatory protein of absorptive epithelial brush borders that organizes the apical actin cytoskeleton and maintains epithelial barrier integrity [#0, #2, #3]. Its F-actin binding is mediated through Cys624: covalent modification of this residue by an enterotoxin triggers F-actin disassembly and barrier failure, while a protective small molecule that shields Cys624 allosterically enhances F-actin binding affinity and preserves barrier integrity [#2]. Across absorptive cell types VIL1 (and its fish ortholog VILL) is required for formation of apical microvilli [#3]. In intestinal epithelial cells, VIL1 together with gelsolin is required for dephosphorylation of EIF2A and recovery from cell stress; their chronic loss produces constitutive EIF2A phosphorylation and integrated stress response activation, IRGM1 overexpression, and RIPK3-mediated necroptotic death, with double-knockout mice developing spontaneous Crohn's-like ileitis that is reversed by ISR or RIPK3 inhibition [#0, #5]. In colorectal cancer cells VIL1 suppresses ferroptosis and promotes tumor growth by binding NF-\\u03baB p105 and driving NF-\\u03baB and lipocalin-2 (LCN2) expression [#1]. Release of VIL1 from injured brush-border cells into plasma tracks the severity of necroptotic proximal tubular injury [#4].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 2014,\n      \"claim\": \"Establishing whether villin-1 actively builds apical microvilli rather than merely decorating them, loss-of-function in a fish ortholog tied VILL directly to actin-based microvillus formation in absorptive cells.\",\n      \"evidence\": \"Morpholino knockdown of VILL in medaka gill ionocytes with immunofluorescence and scanning electron microscopy across salinity conditions\",\n      \"pmids\": [\"24410933\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Ortholog in a fish model, not direct human VIL1\", \"Does not define the molecular mechanism by which VILL nucleates or bundles apical actin\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Connecting brush-border actin regulators to epithelial stress recovery, VIL1 and gelsolin were shown to be required for EIF2A dephosphorylation, linking their loss to constitutive stress signaling, necroptosis, and Crohn's-like inflammation.\",\n      \"evidence\": \"Villin-1/gelsolin double-knockout mice with histology, immunoblots, flow cytometry, electron microscopy, and corroboration in human CD tissue\",\n      \"pmids\": [\"29274870\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular link between VIL1 and the EIF2A phosphatase machinery not resolved\", \"Whether the effect requires VIL1's actin-binding activity is untested\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Testing whether VIL1 release reports epithelial death, plasmatic villin-1 was shown to rise with proximal tubular injury and to depend on necroptosis, validating it as a necroptosis-linked injury marker.\",\n      \"evidence\": \"Immunohistochemistry and Western blotting across rat, pig, and human renal/liver injury models with necrostatin-1 intervention\",\n      \"pmids\": [\"28704336\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of VIL1 redistribution and release not defined\", \"Single lab; biomarker specificity versus other brush-border proteins unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Asking how VIL1 influences tumor cell fate, knockout/overexpression in colorectal cancer cells placed VIL1 upstream of ferroptosis suppression via direct NF-\\u03baB p105 binding and LCN2 induction.\",\n      \"evidence\": \"VIL1 knockout/overexpression in CRC lines with xenografts, transcriptomics, immunoblotting, and ferroptosis assays\",\n      \"pmids\": [\"39658274\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct VIL1\\u2013NF-\\u03baB binding interface not mapped\", \"Relationship between VIL1's actin role and NF-\\u03baB regulation unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identifying the molecular site of VIL1's actin control, Cys624 was shown to be the covalently targetable residue governing F-actin binding and barrier integrity.\",\n      \"evidence\": \"Intestinal organoids, chemical proteomics, Cys624 mutagenesis, in vitro F-actin binding assays, and in vivo barrier validation\",\n      \"pmids\": [\"40835194\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of Cys624-dependent actin binding not resolved\", \"Whether physiological modification of Cys624 occurs endogenously is unknown\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Resolving the causal hierarchy of VIL1-loss pathology, ISR and RIPK3 were placed downstream of VIL1 loss, with pharmacologic inhibition rescuing epithelial architecture and barrier function.\",\n      \"evidence\": \"Villin-1/gelsolin double-knockout mice and CD patient-derived enteroids with ISR/RIPK3 inhibitor rescue and transepithelial resistance readouts\",\n      \"pmids\": [\"42254860\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical step connecting VIL1 loss to ISR activation still undefined\", \"Single lab extending prior findings\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How VIL1's actin-bundling activity at the brush border mechanistically couples to its roles in EIF2A dephosphorylation and NF-\\u03baB binding remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model linking actin binding to stress-signaling functions\", \"Whether actin-independent scaffolding underlies the NF-\\u03baB and ISR roles is untested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [2, 3]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3, 4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [0, 5]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 5]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"GSN\", \"ACTB\", \"NFKB1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win"}}