{"gene":"EVPL","run_date":"2026-04-28T17:46:03","timeline":{"discoveries":[{"year":1996,"finding":"Envoplakin (EVPL) is a membrane-associated precursor of the epidermal cornified envelope, homologous to desmoplakin I/II, BPAG1, and plectin, and is proposed to link desmosomes and keratin filaments to the cornified envelope. The human EVPL gene was localized to chromosome 17q25 by somatic cell hybrid analysis and chromosomal in situ hybridisation.","method":"Somatic cell hybrid analysis, chromosomal in situ hybridisation, sequence homology analysis","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 2 — direct chromosomal localization experiment; functional role proposed based on structural homology with characterized family members","pmids":["8938451"],"is_preprint":false},{"year":2005,"finding":"p63 directly regulates EVPL expression through binding to a p63-specific response element (RRRCGTGYYY) in the EVPL promoter, distinct from the canonical p53-RE. EVPL protein is highly expressed in epithelial cells of skin and pharynx in p63+/+ mice but is undetectable in p63-/- mice, establishing p63 as a transcriptional activator of EVPL.","method":"Oligonucleotide expression microarray, luciferase reporter assays with mutagenized promoter constructs, electromobility shift assay (EMSA), p63 knockout mouse immunostaining","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods including EMSA, mutagenized promoter assays, and in vivo knockout validation","pmids":["15988020"],"is_preprint":false},{"year":2016,"finding":"Autoantibodies in paraneoplastic pemphigus (PNP) patients recognize EVPL at two major epitope regions: EVPL-N1 (aa1–141, N-terminus) and EVPL-L3 (aa1684–1784, C-terminus of the linker subdomain), each with 75.38% sensitivity for PNP detection. Autoantibodies against EVPL-N3 were associated with bronchiolitis obliterans or lichen planus-like lesions.","method":"ELISA using 12 overlapping truncated recombinant fragments of EVPL expressed in E. coli, tested against sera from 65 PNP patients and 50 healthy controls","journal":"Journal of dermatological science","confidence":"Medium","confidence_rationale":"Tier 2 — systematic epitope mapping with recombinant protein fragments and large patient cohort; identifies functional antigenic domains","pmids":["27427435"],"is_preprint":false},{"year":2021,"finding":"Knockdown of EVPL in ESCC (KYSE-150) cell lines led to increased radioresistance. Transcriptome analysis indicated that EVPL regulates expression of interleukins, interleukin receptors, and chemokines by modulating the NF-κB and TNF signaling pathways in radioresistant ESCC cells, thereby suppressing immune response.","method":"RNAi knockdown in ESCC cell lines, clonogenic survival assay after irradiation, RNA sequencing of knockdown cells","journal":"OncoTargets and therapy","confidence":"Medium","confidence_rationale":"Tier 2 — direct loss-of-function with defined cellular phenotype and transcriptomic pathway analysis; single lab study","pmids":["33633453"],"is_preprint":false},{"year":2024,"finding":"EVPL inhibits melanoma cell proliferation, migration, and invasion, and suppresses macrophage recruitment and polarization through the RAS/ERK signaling pathway. Overexpression of EVPL suppressed malignant progression, while EVPL knockdown reversed the inhibitory effects of the ERK inhibitor SCH772984. EVPL inhibits chemokine expression and macrophage M2 polarization via RAS/RAF/ERK signaling.","method":"CCK-8, colony formation, wound healing, Transwell invasion, flow cytometry, ELISA, co-culture Transwell system, Western blot for pathway proteins, EVPL overexpression and shRNA knockdown in melanoma cells","journal":"Journal of inflammation research","confidence":"Medium","confidence_rationale":"Tier 2 — multiple in vitro functional assays with rescue/inhibitor experiments placing EVPL upstream of RAS/ERK; single lab","pmids":["38504693"],"is_preprint":false},{"year":2024,"finding":"BRD4 physically interacts with p63 (and other p53 family members), and this interaction sustains transcription of EVPL in keratinocytes. ChIP and transcriptomic analyses showed BRD4 and p63 act synergistically at the EVPL locus; depletion of either p63 or BRD4, or BRD4 inhibition with JQ1, reduced EVPL expression and altered keratinocyte differentiation.","method":"Protein-protein interaction assay, chromatin immunoprecipitation (ChIP), RNA-seq, RT-qPCR, BRD4 inhibitor (JQ1) treatment, p63/BRD4 knockdown in keratinocytes","journal":"Biology direct","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP/interaction plus ChIP and transcriptomic validation; single lab but multiple orthogonal methods","pmids":["39605045"],"is_preprint":false},{"year":2026,"finding":"AGTPBP1 (a cytosolic carboxypeptidase) negatively regulates EVPL expression in breast cancer cells. AGTPBP1-mediated repression of EVPL leads to activation of ERK signaling (increased ERK1/2 phosphorylation). Rescue experiments showed that EVPL overexpression attenuated AGTPBP1-induced malignant phenotypes (proliferation, migration, invasion) and ERK activation, establishing an AGTPBP1/EVPL/ERK signaling axis.","method":"RNA sequencing, stable overexpression and knockdown of AGTPBP1, EVPL overexpression rescue experiments, Western blot for p-ERK1/2, proliferation/colony formation/migration/invasion assays","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 — RNA-seq plus rescue experiments with defined pathway placement; single lab with multiple orthogonal methods","pmids":["41905574"],"is_preprint":false}],"current_model":"EVPL (envoplakin) is a plakin family cytolinker protein that functions as a membrane-associated precursor of the epidermal cornified envelope, linking desmosomes and keratin filaments; its transcription is directly activated by p63 (and co-regulated by BRD4) through a specific promoter response element; EVPL acts as a tumor suppressor in epithelial cancers by inhibiting ERK signaling (downstream of RAS/RAF) and modulating NF-κB/TNF pathways, and EVPL loss leads to radioresistance and increased malignant progression, while EVPL is also a major autoantigen in paraneoplastic pemphigus with defined epitope regions at its N-terminus and linker subdomain."},"narrative":{"teleology":[{"year":1996,"claim":"Identification of EVPL as a plakin family member and cornified envelope precursor established its structural kinship with desmoplakin, BPAG1, and plectin, suggesting a cytolinker function bridging desmosomes to keratin filaments.","evidence":"Somatic cell hybrid/FISH mapping to 17q25 and sequence homology analysis","pmids":["8938451"],"confidence":"Medium","gaps":["Functional cytolinker role inferred from homology, not directly demonstrated","No direct interaction data with desmosomal or intermediate filament proteins"]},{"year":2005,"claim":"Discovery that p63 directly transactivates EVPL through a non-canonical response element resolved how EVPL expression is restricted to stratified epithelia and connected it to the master epithelial transcription factor p63.","evidence":"EMSA, luciferase reporter mutagenesis, and loss of EVPL protein in p63-knockout mouse skin","pmids":["15988020"],"confidence":"High","gaps":["Whether other transcription factors cooperate with p63 at the EVPL promoter was unknown at this stage","Post-transcriptional regulation of EVPL unexplored"]},{"year":2016,"claim":"Systematic epitope mapping of EVPL in paraneoplastic pemphigus defined two dominant autoantibody-binding regions, linking specific EVPL structural domains to autoimmune pathology.","evidence":"ELISA with 12 overlapping recombinant EVPL fragments tested against 65 PNP patient sera","pmids":["27427435"],"confidence":"Medium","gaps":["Whether autoantibodies are pathogenic (causing tissue damage) versus epiphenomenal not established","Structural basis of epitope immunodominance unknown"]},{"year":2021,"claim":"Demonstrating that EVPL knockdown increases radioresistance and alters NF-κB/TNF-dependent cytokine expression first connected EVPL to immune signaling and treatment response in cancer.","evidence":"RNAi knockdown in ESCC cells with clonogenic survival assays and RNA-seq","pmids":["33633453"],"confidence":"Medium","gaps":["Mechanism by which EVPL modulates NF-κB/TNF signaling not defined","In vivo validation of radioresistance phenotype absent","Single cell line tested"]},{"year":2024,"claim":"Placing EVPL upstream of RAS/RAF/ERK signaling in melanoma, with rescue and inhibitor experiments, established EVPL as a tumor suppressor that restrains ERK-driven proliferation, invasion, and macrophage polarization.","evidence":"EVPL overexpression/knockdown in melanoma cells with ERK inhibitor rescue, co-culture Transwell, Western blot for pathway components","pmids":["38504693"],"confidence":"Medium","gaps":["Direct molecular target through which EVPL inhibits RAS/ERK not identified","In vivo tumor suppressor activity not demonstrated","Whether EVPL acts at the membrane, cytosol, or both in this context is unclear"]},{"year":2024,"claim":"Identification of BRD4 as a p63 co-activator at the EVPL locus refined the transcriptional regulatory mechanism and linked epigenetic reader activity to EVPL-dependent keratinocyte differentiation.","evidence":"BRD4–p63 co-immunoprecipitation, ChIP at EVPL locus, JQ1 inhibitor and knockdown experiments in keratinocytes","pmids":["39605045"],"confidence":"Medium","gaps":["Whether BRD4 is required specifically at the EVPL locus versus broadly for p63 target genes not distinguished","Functional consequence of BRD4-mediated EVPL regulation on cornified envelope assembly not tested"]},{"year":2026,"claim":"Discovery of the AGTPBP1/EVPL/ERK axis in breast cancer provided an upstream repressor of EVPL and reinforced the generality of the EVPL–ERK tumor suppressor mechanism across cancer types.","evidence":"RNA-seq, AGTPBP1 overexpression/knockdown with EVPL rescue, Western blot for p-ERK1/2 in breast cancer cells","pmids":["41905574"],"confidence":"Medium","gaps":["Mechanism by which AGTPBP1 (a carboxypeptidase) represses EVPL expression is unknown","Whether EVPL directly binds a RAS/ERK pathway component remains unresolved","In vivo validation of this axis absent"]},{"year":null,"claim":"The direct molecular mechanism by which EVPL suppresses RAS/ERK signaling — whether through protein–protein interaction, scaffolding, or indirect effects — remains unknown, as does EVPL's precise structural role in cornified envelope assembly beyond homology-based inference.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No binding partner or direct target in the RAS/ERK pathway identified for EVPL","Structural/biochemical reconstitution of EVPL in cornified envelope assembly not performed","In vivo tumor suppressor function of EVPL not demonstrated in animal models"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[3,4,6]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[4,6]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,4,6]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[1,5]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[3,4,6]}],"complexes":[],"partners":["TP63","BRD4","AGTPBP1"],"other_free_text":[]},"mechanistic_narrative":"EVPL (envoplakin) is a plakin family cytolinker protein that serves as a membrane-associated precursor of the epidermal cornified envelope, linking desmosomes and keratin intermediate filaments in stratified epithelia [PMID:8938451]. Its transcription is directly activated by p63 through a specific response element (RRRCGTGYYY) in the EVPL promoter, with BRD4 acting as a co-activator of p63-dependent EVPL expression in keratinocytes [PMID:15988020, PMID:39605045]. EVPL functions as a tumor suppressor by inhibiting the RAS/RAF/ERK signaling pathway, suppressing cancer cell proliferation, migration, invasion, and macrophage M2 polarization; its loss also confers radioresistance and modulates NF-κB/TNF-dependent immune gene expression in esophageal squamous cell carcinoma [PMID:38504693, PMID:33633453, PMID:41905574]. EVPL is a major autoantigen in paraneoplastic pemphigus, with dominant B-cell epitopes mapped to its N-terminal domain (aa 1–141) and linker subdomain (aa 1684–1784) [PMID:27427435]."},"prefetch_data":{"uniprot":{"accession":"Q92817","full_name":"Envoplakin","aliases":["210 kDa cornified envelope precursor protein","210 kDa paraneoplastic pemphigus antigen","p210"],"length_aa":2033,"mass_kda":231.6,"function":"Component of the cornified envelope of keratinocytes. May link the cornified envelope to desmosomes and intermediate filaments","subcellular_location":"Cell junction, desmosome; Cornified envelope; Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/Q92817/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EVPL","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"AATF","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/EVPL","total_profiled":1310},"omim":[{"mim_id":"618301","title":"KAZRIN, PERIPLAKIN-INTERACTING PROTEIN; KAZN","url":"https://www.omim.org/entry/618301"},{"mim_id":"601590","title":"ENVOPLAKIN; EVPL","url":"https://www.omim.org/entry/601590"},{"mim_id":"601041","title":"TLE FAMILY MEMBER 2, TRANSCRIPTIONAL COREPRESSOR; TLE2","url":"https://www.omim.org/entry/601041"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Intermediate filaments","reliability":"Supported"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"cervix","ntpm":90.8},{"tissue":"esophagus","ntpm":289.2},{"tissue":"skin 1","ntpm":148.1},{"tissue":"vagina","ntpm":102.1}],"url":"https://www.proteinatlas.org/search/EVPL"},"hgnc":{"alias_symbol":["EVPK"],"prev_symbol":[]},"alphafold":{"accession":"Q92817","domains":[{"cath_id":"1.20.58.60","chopping":"37-140","consensus_level":"medium","plddt":87.1317,"start":37,"end":140},{"cath_id":"2.30.30.40","chopping":"308-361_376-502","consensus_level":"medium","plddt":87.3663,"start":308,"end":502},{"cath_id":"1.20.58.60","chopping":"524-745","consensus_level":"high","plddt":87.3585,"start":524,"end":745},{"cath_id":"1.20.58,1.20.58","chopping":"748-838_849-885","consensus_level":"medium","plddt":83.7561,"start":748,"end":885},{"cath_id":"3.90.1290.10","chopping":"1824-2014","consensus_level":"high","plddt":84.9814,"start":1824,"end":2014}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q92817","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q92817-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q92817-F1-predicted_aligned_error_v6.png","plddt_mean":71.69},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EVPL","jax_strain_url":"https://www.jax.org/strain/search?query=EVPL"},"sequence":{"accession":"Q92817","fasta_url":"https://rest.uniprot.org/uniprotkb/Q92817.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q92817/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q92817"}},"corpus_meta":[{"pmid":"17986187","id":"PMC_17986187","title":"Dissection 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BPAG1, and plectin, and is proposed to link desmosomes and keratin filaments to the cornified envelope. The human EVPL gene was localized to chromosome 17q25 by somatic cell hybrid analysis and chromosomal in situ hybridisation.\",\n      \"method\": \"Somatic cell hybrid analysis, chromosomal in situ hybridisation, sequence homology analysis\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct chromosomal localization experiment; functional role proposed based on structural homology with characterized family members\",\n      \"pmids\": [\"8938451\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"p63 directly regulates EVPL expression through binding to a p63-specific response element (RRRCGTGYYY) in the EVPL promoter, distinct from the canonical p53-RE. EVPL protein is highly expressed in epithelial cells of skin and pharynx in p63+/+ mice but is undetectable in p63-/- mice, establishing p63 as a transcriptional activator of EVPL.\",\n      \"method\": \"Oligonucleotide expression microarray, luciferase reporter assays with mutagenized promoter constructs, electromobility shift assay (EMSA), p63 knockout mouse immunostaining\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods including EMSA, mutagenized promoter assays, and in vivo knockout validation\",\n      \"pmids\": [\"15988020\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Autoantibodies in paraneoplastic pemphigus (PNP) patients recognize EVPL at two major epitope regions: EVPL-N1 (aa1–141, N-terminus) and EVPL-L3 (aa1684–1784, C-terminus of the linker subdomain), each with 75.38% sensitivity for PNP detection. Autoantibodies against EVPL-N3 were associated with bronchiolitis obliterans or lichen planus-like lesions.\",\n      \"method\": \"ELISA using 12 overlapping truncated recombinant fragments of EVPL expressed in E. coli, tested against sera from 65 PNP patients and 50 healthy controls\",\n      \"journal\": \"Journal of dermatological science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — systematic epitope mapping with recombinant protein fragments and large patient cohort; identifies functional antigenic domains\",\n      \"pmids\": [\"27427435\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Knockdown of EVPL in ESCC (KYSE-150) cell lines led to increased radioresistance. Transcriptome analysis indicated that EVPL regulates expression of interleukins, interleukin receptors, and chemokines by modulating the NF-κB and TNF signaling pathways in radioresistant ESCC cells, thereby suppressing immune response.\",\n      \"method\": \"RNAi knockdown in ESCC cell lines, clonogenic survival assay after irradiation, RNA sequencing of knockdown cells\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct loss-of-function with defined cellular phenotype and transcriptomic pathway analysis; single lab study\",\n      \"pmids\": [\"33633453\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EVPL inhibits melanoma cell proliferation, migration, and invasion, and suppresses macrophage recruitment and polarization through the RAS/ERK signaling pathway. Overexpression of EVPL suppressed malignant progression, while EVPL knockdown reversed the inhibitory effects of the ERK inhibitor SCH772984. EVPL inhibits chemokine expression and macrophage M2 polarization via RAS/RAF/ERK signaling.\",\n      \"method\": \"CCK-8, colony formation, wound healing, Transwell invasion, flow cytometry, ELISA, co-culture Transwell system, Western blot for pathway proteins, EVPL overexpression and shRNA knockdown in melanoma cells\",\n      \"journal\": \"Journal of inflammation research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple in vitro functional assays with rescue/inhibitor experiments placing EVPL upstream of RAS/ERK; single lab\",\n      \"pmids\": [\"38504693\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"BRD4 physically interacts with p63 (and other p53 family members), and this interaction sustains transcription of EVPL in keratinocytes. ChIP and transcriptomic analyses showed BRD4 and p63 act synergistically at the EVPL locus; depletion of either p63 or BRD4, or BRD4 inhibition with JQ1, reduced EVPL expression and altered keratinocyte differentiation.\",\n      \"method\": \"Protein-protein interaction assay, chromatin immunoprecipitation (ChIP), RNA-seq, RT-qPCR, BRD4 inhibitor (JQ1) treatment, p63/BRD4 knockdown in keratinocytes\",\n      \"journal\": \"Biology direct\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP/interaction plus ChIP and transcriptomic validation; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"39605045\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"AGTPBP1 (a cytosolic carboxypeptidase) negatively regulates EVPL expression in breast cancer cells. AGTPBP1-mediated repression of EVPL leads to activation of ERK signaling (increased ERK1/2 phosphorylation). Rescue experiments showed that EVPL overexpression attenuated AGTPBP1-induced malignant phenotypes (proliferation, migration, invasion) and ERK activation, establishing an AGTPBP1/EVPL/ERK signaling axis.\",\n      \"method\": \"RNA sequencing, stable overexpression and knockdown of AGTPBP1, EVPL overexpression rescue experiments, Western blot for p-ERK1/2, proliferation/colony formation/migration/invasion assays\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — RNA-seq plus rescue experiments with defined pathway placement; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"41905574\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EVPL (envoplakin) is a plakin family cytolinker protein that functions as a membrane-associated precursor of the epidermal cornified envelope, linking desmosomes and keratin filaments; its transcription is directly activated by p63 (and co-regulated by BRD4) through a specific promoter response element; EVPL acts as a tumor suppressor in epithelial cancers by inhibiting ERK signaling (downstream of RAS/RAF) and modulating NF-κB/TNF pathways, and EVPL loss leads to radioresistance and increased malignant progression, while EVPL is also a major autoantigen in paraneoplastic pemphigus with defined epitope regions at its N-terminus and linker subdomain.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"EVPL (envoplakin) is a plakin family cytolinker protein that serves as a membrane-associated precursor of the epidermal cornified envelope, linking desmosomes and keratin intermediate filaments in stratified epithelia [PMID:8938451]. Its transcription is directly activated by p63 through a specific response element (RRRCGTGYYY) in the EVPL promoter, with BRD4 acting as a co-activator of p63-dependent EVPL expression in keratinocytes [PMID:15988020, PMID:39605045]. EVPL functions as a tumor suppressor by inhibiting the RAS/RAF/ERK signaling pathway, suppressing cancer cell proliferation, migration, invasion, and macrophage M2 polarization; its loss also confers radioresistance and modulates NF-κB/TNF-dependent immune gene expression in esophageal squamous cell carcinoma [PMID:38504693, PMID:33633453, PMID:41905574]. EVPL is a major autoantigen in paraneoplastic pemphigus, with dominant B-cell epitopes mapped to its N-terminal domain (aa 1–141) and linker subdomain (aa 1684–1784) [PMID:27427435].\",\n  \"teleology\": [\n    {\n      \"year\": 1996,\n      \"claim\": \"Identification of EVPL as a plakin family member and cornified envelope precursor established its structural kinship with desmoplakin, BPAG1, and plectin, suggesting a cytolinker function bridging desmosomes to keratin filaments.\",\n      \"evidence\": \"Somatic cell hybrid/FISH mapping to 17q25 and sequence homology analysis\",\n      \"pmids\": [\"8938451\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Functional cytolinker role inferred from homology, not directly demonstrated\",\n        \"No direct interaction data with desmosomal or intermediate filament proteins\"\n      ]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Discovery that p63 directly transactivates EVPL through a non-canonical response element resolved how EVPL expression is restricted to stratified epithelia and connected it to the master epithelial transcription factor p63.\",\n      \"evidence\": \"EMSA, luciferase reporter mutagenesis, and loss of EVPL protein in p63-knockout mouse skin\",\n      \"pmids\": [\"15988020\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether other transcription factors cooperate with p63 at the EVPL promoter was unknown at this stage\",\n        \"Post-transcriptional regulation of EVPL unexplored\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Systematic epitope mapping of EVPL in paraneoplastic pemphigus defined two dominant autoantibody-binding regions, linking specific EVPL structural domains to autoimmune pathology.\",\n      \"evidence\": \"ELISA with 12 overlapping recombinant EVPL fragments tested against 65 PNP patient sera\",\n      \"pmids\": [\"27427435\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether autoantibodies are pathogenic (causing tissue damage) versus epiphenomenal not established\",\n        \"Structural basis of epitope immunodominance unknown\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrating that EVPL knockdown increases radioresistance and alters NF-κB/TNF-dependent cytokine expression first connected EVPL to immune signaling and treatment response in cancer.\",\n      \"evidence\": \"RNAi knockdown in ESCC cells with clonogenic survival assays and RNA-seq\",\n      \"pmids\": [\"33633453\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which EVPL modulates NF-κB/TNF signaling not defined\",\n        \"In vivo validation of radioresistance phenotype absent\",\n        \"Single cell line tested\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placing EVPL upstream of RAS/RAF/ERK signaling in melanoma, with rescue and inhibitor experiments, established EVPL as a tumor suppressor that restrains ERK-driven proliferation, invasion, and macrophage polarization.\",\n      \"evidence\": \"EVPL overexpression/knockdown in melanoma cells with ERK inhibitor rescue, co-culture Transwell, Western blot for pathway components\",\n      \"pmids\": [\"38504693\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct molecular target through which EVPL inhibits RAS/ERK not identified\",\n        \"In vivo tumor suppressor activity not demonstrated\",\n        \"Whether EVPL acts at the membrane, cytosol, or both in this context is unclear\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identification of BRD4 as a p63 co-activator at the EVPL locus refined the transcriptional regulatory mechanism and linked epigenetic reader activity to EVPL-dependent keratinocyte differentiation.\",\n      \"evidence\": \"BRD4–p63 co-immunoprecipitation, ChIP at EVPL locus, JQ1 inhibitor and knockdown experiments in keratinocytes\",\n      \"pmids\": [\"39605045\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether BRD4 is required specifically at the EVPL locus versus broadly for p63 target genes not distinguished\",\n        \"Functional consequence of BRD4-mediated EVPL regulation on cornified envelope assembly not tested\"\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Discovery of the AGTPBP1/EVPL/ERK axis in breast cancer provided an upstream repressor of EVPL and reinforced the generality of the EVPL–ERK tumor suppressor mechanism across cancer types.\",\n      \"evidence\": \"RNA-seq, AGTPBP1 overexpression/knockdown with EVPL rescue, Western blot for p-ERK1/2 in breast cancer cells\",\n      \"pmids\": [\"41905574\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which AGTPBP1 (a carboxypeptidase) represses EVPL expression is unknown\",\n        \"Whether EVPL directly binds a RAS/ERK pathway component remains unresolved\",\n        \"In vivo validation of this axis absent\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The direct molecular mechanism by which EVPL suppresses RAS/ERK signaling — whether through protein–protein interaction, scaffolding, or indirect effects — remains unknown, as does EVPL's precise structural role in cornified envelope assembly beyond homology-based inference.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No binding partner or direct target in the RAS/ERK pathway identified for EVPL\",\n        \"Structural/biochemical reconstitution of EVPL in cornified envelope assembly not performed\",\n        \"In vivo tumor suppressor function of EVPL not demonstrated in animal models\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [3, 4, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [4, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 4, 6]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [1, 5]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [3, 4, 6]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"TP63\",\n      \"BRD4\",\n      \"AGTPBP1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}