{"gene":"EVPL","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":1996,"finding":"Envoplakin (EVPL) is a membrane-associated precursor of the epidermal cornified envelope, homologous to desmoplakin, BPAG1, and plectin, and 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, cosmid/YAC isolation","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct chromosomal localization by two orthogonal mapping methods; functional role proposed based on homology and expression pattern, not directly experimentally validated in this paper","pmids":["8938451"],"is_preprint":false},{"year":2005,"finding":"p63 directly regulates EVPL expression through a p63-specific response element (p63-RE: RRRCGTGYYY) in the EVPL promoter that differs from the canonical p53-RE. Electromobility shift assays and luciferase assays on mutagenized promoter constructs showed p63 preferentially binds and activates RRRCGTGYYY, whereas p53 preferentially activates RRRCATGYYY. EVPL protein is highly expressed in epithelial cells of skin and pharynx in p63+/+ mice but undetectable in p63-/- mice.","method":"Oligonucleotide expression microarray, luciferase assays on mutagenized promoter constructs, electromobility shift assays (EMSA), p63 knockout mouse immunostaining","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal methods (mutagenized promoter luciferase, EMSA, in vivo knockout validation) in a single rigorous study","pmids":["15988020"],"is_preprint":false},{"year":1999,"finding":"Mutation analysis of EVPL in three tylosis esophageal cancer (TOC) families failed to identify tylosis-specific mutations. Haplotype analysis of three intragenic EVPL polymorphisms placed EVPL proximal to D17S1839, outside the minimal TOC region, excluding EVPL as the TOC causative gene.","method":"Mutation analysis, haplotype analysis, radiation hybrid mapping, YAC/BAC/PAC physical mapping","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — integrated genetic and physical mapping with multiple markers; negative result well-supported by orthogonal mapping approaches","pmids":["10409435"],"is_preprint":false},{"year":2005,"finding":"Somatic EVPL gene mutation rate was very low (5%, 1/20) in sporadic esophageal squamous cell carcinoma (OSC), despite frequent LOH (63%) at the TOC locus, suggesting EVPL is not the primary target gene responsible for OSC at this locus. One somatic missense mutation (Ala1104Thr) was found in the central rod domain.","method":"RT-PCR SSCP and direct sequencing of EVPL in OSC cell lines and tumor samples","journal":"Oncology reports","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct sequencing of tumor samples; negative/low-frequency result for EVPL mutation, single lab","pmids":["15756445"],"is_preprint":false},{"year":2016,"finding":"Autoantibody epitopes of EVPL in paraneoplastic pemphigus (PNP) are concentrated in the N-terminal fragment EVPL-N1 (aa1-141) and the linker subdomain fragment EVPL-L3 (aa1684-1784), each recognized at 75.38% sensitivity in PNP patient sera. Autoantibodies against EVPL-N3 were associated with PNP patients with bronchiolitis obliterans or lichen planus-like lesions.","method":"Production of 12 overlapping truncated EVPL fragments in E. coli; ELISA of PNP patient and control sera against each fragment","journal":"Journal of dermatological science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — systematic epitope mapping with multiple recombinant fragments across 65 PNP patients and 50 controls, single lab","pmids":["27427435"],"is_preprint":false},{"year":2021,"finding":"Knockdown of EVPL in ESCC cell line KYSE-150 led to increased radioresistance. Transcriptome analysis indicated that EVPL knockdown may regulate expression of interleukins, interleukin receptors, and chemokines by inhibiting NF-κB and TNF signaling pathways, thereby suppressing immune response in radioresistant ESCC cells.","method":"RNAi gene knockdown in KYSE-150 ESCC cell line, clonogenic survival assay, RNA sequencing","journal":"OncoTargets and therapy","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway inference from transcriptome without direct mechanistic validation of NF-κB/TNF pathway involvement","pmids":["33633453"],"is_preprint":false},{"year":2024,"finding":"BRD4 physically interacts with p63 and synergistically co-regulates transcription of EVPL (among other keratinocyte-specific p63 target genes). ChIP and RT-qPCR confirmed BRD4 occupancy at p63 target loci including EVPL. Depletion of BRD4 or inhibition with JQ1 altered EVPL expression alongside keratinocyte differentiation.","method":"Protein-protein interaction assays, chromatin immunoprecipitation (ChIP), RT-qPCR, transcriptomic analysis, BRD4 inhibitor (JQ1) treatment, BRD4/p63 knockdown","journal":"Biology direct","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (Co-IP, ChIP, transcriptomics, pharmacological inhibition) in single lab study","pmids":["39605045"],"is_preprint":false},{"year":2024,"finding":"EVPL overexpression suppressed melanoma cell proliferation, migration, invasion, and promoted apoptosis. EVPL inhibited RAS/ERK signaling, chemokine expression, macrophage recruitment, and macrophage M2 polarization. Inhibition of EVPL expression reversed the tumor-suppressive effects of ERK inhibitor SCH772984, placing EVPL upstream of ERK in this pathway.","method":"CCK-8, colony formation, wound healing, Transwell invasion assays, flow cytometry, ELISA, Transwell co-culture macrophage recruitment assay, Western blot for pathway proteins, EVPL overexpression and shRNA knockdown","journal":"Journal of inflammation research","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — multiple in vitro functional assays and Western blot pathway validation, single lab","pmids":["38504693"],"is_preprint":false},{"year":2026,"finding":"AGTPBP1 negatively regulates EVPL expression in breast cancer cells. AGTPBP1-mediated repression of EVPL leads to activation of the ERK signaling pathway (increased ERK1/2 phosphorylation). Rescue experiments confirmed that EVPL overexpression attenuated AGTPBP1-induced malignant phenotypes (proliferation, migration, invasion) and ERK activation, establishing an AGTPBP1/EVPL/ERK axis.","method":"RNA sequencing (transcriptomic profiling), EVPL/AGTPBP1 overexpression and knockdown, proliferation/colony formation/migration/invasion assays, Western blot for pERK1/2, rescue experiments","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNA-seq plus functional rescue experiments in two cell lines; single lab but multiple orthogonal methods","pmids":["41905574"],"is_preprint":false}],"current_model":"EVPL (envoplakin) is a plakin-family desmosomal/cornified envelope protein whose transcription is directly regulated by p63 via a p63-specific promoter response element (with BRD4 as a co-activator), and which functions as a structural linker between desmosomes and keratin filaments; loss of EVPL activates the RAS/ERK signaling pathway, promoting cell proliferation, migration, macrophage recruitment, and radioresistance, while EVPL overexpression suppresses these malignant behaviors, indicating that EVPL acts as a tumor suppressor upstream of ERK in epithelial and melanoma contexts."},"narrative":{"mechanistic_narrative":"EVPL (envoplakin) is a plakin-family protein and a membrane-associated precursor of the epidermal cornified envelope, proposed on the basis of homology to desmoplakin, BPAG1, and plectin to link desmosomes and keratin filaments to the cornified envelope [PMID:8938451]. Its expression in epithelial tissues is directly driven by the transcription factor p63, which binds a p63-specific response element (RRRCGTGYYY) in the EVPL promoter that is distinct from the canonical p53 element; EVPL protein is abundant in skin and pharyngeal epithelium of wild-type mice but absent in p63-null mice [PMID:15988020]. This p63-driven transcription is potentiated by BRD4, which physically interacts with p63 and co-occupies the EVPL locus to coordinate EVPL expression during keratinocyte differentiation [PMID:39605045]. Beyond its structural role, EVPL acts as a tumor suppressor that restrains the RAS/ERK signaling axis: EVPL overexpression suppresses melanoma cell proliferation, migration, and invasion, promotes apoptosis, and dampens chemokine-driven macrophage recruitment and M2 polarization, with epistasis experiments placing EVPL upstream of ERK [PMID:38504693]. In breast cancer cells EVPL is repressed by AGTPBP1, and restoring EVPL attenuates AGTPBP1-induced ERK activation and malignancy, defining an AGTPBP1/EVPL/ERK axis [PMID:41905574]. Although EVPL maps within the tylosis esophageal cancer locus on 17q25, mutation and haplotype analyses excluded it as the causative TOC gene and found only rare somatic mutation in sporadic esophageal carcinoma [PMID:10409435, PMID:15756445].","teleology":[{"year":1996,"claim":"Established the molecular identity and candidate function of EVPL by placing it in the plakin family and mapping its locus, framing it as a structural connector of desmosomes, keratin filaments, and the cornified envelope.","evidence":"Somatic cell hybrid analysis and in situ hybridisation with cosmid/YAC isolation, plus homology to desmoplakin/BPAG1/plectin","pmids":["8938451"],"confidence":"Medium","gaps":["Structural linker role inferred from homology and expression, not directly tested","No direct demonstration of binding to desmosomal or keratin components"]},{"year":1999,"claim":"Tested whether EVPL is the causative gene at the tylosis esophageal cancer (TOC) locus and excluded it, redirecting the search for the TOC gene.","evidence":"Mutation and haplotype analysis with radiation hybrid and physical mapping across three TOC families","pmids":["10409435"],"confidence":"Medium","gaps":["Negative result does not address whether EVPL contributes to esophageal pathology by other mechanisms","Limited to three families"]},{"year":2005,"claim":"Resolved how EVPL is transcriptionally controlled, showing it is a direct p63 target via a p63-discriminating promoter element, linking EVPL to epithelial differentiation programs.","evidence":"Microarray, mutagenized-promoter luciferase assays, EMSA, and p63 knockout mouse immunostaining","pmids":["15988020"],"confidence":"High","gaps":["Does not establish EVPL's downstream functional consequences","Cofactors of p63 at the EVPL promoter not defined in this study"]},{"year":2005,"claim":"Assessed whether EVPL is somatically targeted in sporadic esophageal squamous cell carcinoma and found mutation is rare despite frequent LOH, arguing against EVPL as the primary driver at the locus.","evidence":"RT-PCR SSCP and direct sequencing of EVPL in OSC cell lines and tumors","pmids":["15756445"],"confidence":"Medium","gaps":["Single missense variant of unknown functional impact","Does not rule out epigenetic or expression-level inactivation"]},{"year":2016,"claim":"Mapped the autoantigenic regions of EVPL in paraneoplastic pemphigus, localizing dominant epitopes to defined N-terminal and linker subdomains.","evidence":"ELISA of patient/control sera against 12 overlapping recombinant EVPL fragments","pmids":["27427435"],"confidence":"Medium","gaps":["Does not address whether autoantibodies are pathogenic","Epitope mapping does not bear on normal EVPL function"]},{"year":2021,"claim":"Linked EVPL loss to therapy resistance, showing EVPL knockdown increases radioresistance in esophageal cancer cells with inferred suppression of inflammatory signaling.","evidence":"RNAi knockdown in KYSE-150 with clonogenic survival assay and RNA-seq","pmids":["33633453"],"confidence":"Low","gaps":["NF-κB/TNF pathway involvement inferred from transcriptome, not directly validated","Single cell line, single lab"]},{"year":2024,"claim":"Identified BRD4 as a transcriptional coactivator partnering with p63 to drive EVPL, refining the regulatory logic of EVPL expression during keratinocyte differentiation.","evidence":"Co-IP, ChIP, RT-qPCR, transcriptomics, and JQ1/knockdown perturbation","pmids":["39605045"],"confidence":"Medium","gaps":["EVPL is one of several co-regulated targets; EVPL-specific dependence on BRD4 not isolated","Direct vs indirect BRD4 effect at EVPL not fully separated"]},{"year":2024,"claim":"Defined a functional tumor-suppressor role for EVPL upstream of ERK, showing EVPL restrains melanoma proliferation, invasion, and macrophage recruitment through RAS/ERK signaling.","evidence":"Gain/loss-of-function in melanoma cells with functional assays, co-culture, and ERK-inhibitor epistasis (SCH772984)","pmids":["38504693"],"confidence":"Medium","gaps":["Molecular mechanism by which EVPL regulates RAS/ERK is undefined","Single lab, in vitro context"]},{"year":2026,"claim":"Placed EVPL within a defined regulatory axis in breast cancer, showing AGTPBP1 represses EVPL to activate ERK and that restoring EVPL reverses malignancy.","evidence":"RNA-seq, gain/loss-of-function, and rescue experiments with pERK1/2 readout in two cell lines","pmids":["41905574"],"confidence":"Medium","gaps":["Mechanism of AGTPBP1-mediated EVPL repression not established","Direct biochemical link between EVPL and ERK pathway components missing"]},{"year":null,"claim":"The molecular mechanism by which EVPL, a structural cornified-envelope protein, suppresses RAS/ERK signaling remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No identified physical link between EVPL and RAS/ERK pathway components","Whether the tumor-suppressive activity depends on EVPL's structural/desmosomal role is unknown","No in vivo validation of the tumor-suppressor function"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[0]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7,8]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[1,6]}],"complexes":[],"partners":[],"other_free_text":[]}},"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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the epidermal cornified envelope, homologous to desmoplakin, BPAG1, and plectin, and 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, cosmid/YAC isolation\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct chromosomal localization by two orthogonal mapping methods; functional role proposed based on homology and expression pattern, not directly experimentally validated in this paper\",\n      \"pmids\": [\"8938451\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"p63 directly regulates EVPL expression through a p63-specific response element (p63-RE: RRRCGTGYYY) in the EVPL promoter that differs from the canonical p53-RE. Electromobility shift assays and luciferase assays on mutagenized promoter constructs showed p63 preferentially binds and activates RRRCGTGYYY, whereas p53 preferentially activates RRRCATGYYY. EVPL protein is highly expressed in epithelial cells of skin and pharynx in p63+/+ mice but undetectable in p63-/- mice.\",\n      \"method\": \"Oligonucleotide expression microarray, luciferase assays on mutagenized promoter constructs, electromobility shift assays (EMSA), p63 knockout mouse immunostaining\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal methods (mutagenized promoter luciferase, EMSA, in vivo knockout validation) in a single rigorous study\",\n      \"pmids\": [\"15988020\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Mutation analysis of EVPL in three tylosis esophageal cancer (TOC) families failed to identify tylosis-specific mutations. Haplotype analysis of three intragenic EVPL polymorphisms placed EVPL proximal to D17S1839, outside the minimal TOC region, excluding EVPL as the TOC causative gene.\",\n      \"method\": \"Mutation analysis, haplotype analysis, radiation hybrid mapping, YAC/BAC/PAC physical mapping\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — integrated genetic and physical mapping with multiple markers; negative result well-supported by orthogonal mapping approaches\",\n      \"pmids\": [\"10409435\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Somatic EVPL gene mutation rate was very low (5%, 1/20) in sporadic esophageal squamous cell carcinoma (OSC), despite frequent LOH (63%) at the TOC locus, suggesting EVPL is not the primary target gene responsible for OSC at this locus. One somatic missense mutation (Ala1104Thr) was found in the central rod domain.\",\n      \"method\": \"RT-PCR SSCP and direct sequencing of EVPL in OSC cell lines and tumor samples\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct sequencing of tumor samples; negative/low-frequency result for EVPL mutation, single lab\",\n      \"pmids\": [\"15756445\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Autoantibody epitopes of EVPL in paraneoplastic pemphigus (PNP) are concentrated in the N-terminal fragment EVPL-N1 (aa1-141) and the linker subdomain fragment EVPL-L3 (aa1684-1784), each recognized at 75.38% sensitivity in PNP patient sera. Autoantibodies against EVPL-N3 were associated with PNP patients with bronchiolitis obliterans or lichen planus-like lesions.\",\n      \"method\": \"Production of 12 overlapping truncated EVPL fragments in E. coli; ELISA of PNP patient and control sera against each fragment\",\n      \"journal\": \"Journal of dermatological science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic epitope mapping with multiple recombinant fragments across 65 PNP patients and 50 controls, single lab\",\n      \"pmids\": [\"27427435\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Knockdown of EVPL in ESCC cell line KYSE-150 led to increased radioresistance. Transcriptome analysis indicated that EVPL knockdown may regulate expression of interleukins, interleukin receptors, and chemokines by inhibiting NF-κB and TNF signaling pathways, thereby suppressing immune response in radioresistant ESCC cells.\",\n      \"method\": \"RNAi gene knockdown in KYSE-150 ESCC cell line, clonogenic survival assay, RNA sequencing\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway inference from transcriptome without direct mechanistic validation of NF-κB/TNF pathway involvement\",\n      \"pmids\": [\"33633453\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"BRD4 physically interacts with p63 and synergistically co-regulates transcription of EVPL (among other keratinocyte-specific p63 target genes). ChIP and RT-qPCR confirmed BRD4 occupancy at p63 target loci including EVPL. Depletion of BRD4 or inhibition with JQ1 altered EVPL expression alongside keratinocyte differentiation.\",\n      \"method\": \"Protein-protein interaction assays, chromatin immunoprecipitation (ChIP), RT-qPCR, transcriptomic analysis, BRD4 inhibitor (JQ1) treatment, BRD4/p63 knockdown\",\n      \"journal\": \"Biology direct\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (Co-IP, ChIP, transcriptomics, pharmacological inhibition) in single lab study\",\n      \"pmids\": [\"39605045\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EVPL overexpression suppressed melanoma cell proliferation, migration, invasion, and promoted apoptosis. EVPL inhibited RAS/ERK signaling, chemokine expression, macrophage recruitment, and macrophage M2 polarization. Inhibition of EVPL expression reversed the tumor-suppressive effects of ERK inhibitor SCH772984, placing EVPL upstream of ERK in this pathway.\",\n      \"method\": \"CCK-8, colony formation, wound healing, Transwell invasion assays, flow cytometry, ELISA, Transwell co-culture macrophage recruitment assay, Western blot for pathway proteins, EVPL overexpression and shRNA knockdown\",\n      \"journal\": \"Journal of inflammation research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — multiple in vitro functional assays and Western blot pathway validation, single lab\",\n      \"pmids\": [\"38504693\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"AGTPBP1 negatively regulates EVPL expression in breast cancer cells. AGTPBP1-mediated repression of EVPL leads to activation of the ERK signaling pathway (increased ERK1/2 phosphorylation). Rescue experiments confirmed that EVPL overexpression attenuated AGTPBP1-induced malignant phenotypes (proliferation, migration, invasion) and ERK activation, establishing an AGTPBP1/EVPL/ERK axis.\",\n      \"method\": \"RNA sequencing (transcriptomic profiling), EVPL/AGTPBP1 overexpression and knockdown, proliferation/colony formation/migration/invasion assays, Western blot for pERK1/2, rescue experiments\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNA-seq plus functional rescue experiments in two cell lines; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"41905574\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EVPL (envoplakin) is a plakin-family desmosomal/cornified envelope protein whose transcription is directly regulated by p63 via a p63-specific promoter response element (with BRD4 as a co-activator), and which functions as a structural linker between desmosomes and keratin filaments; loss of EVPL activates the RAS/ERK signaling pathway, promoting cell proliferation, migration, macrophage recruitment, and radioresistance, while EVPL overexpression suppresses these malignant behaviors, indicating that EVPL acts as a tumor suppressor upstream of ERK in epithelial and melanoma contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EVPL (envoplakin) is a plakin-family protein and a membrane-associated precursor of the epidermal cornified envelope, proposed on the basis of homology to desmoplakin, BPAG1, and plectin to link desmosomes and keratin filaments to the cornified envelope [#0]. Its expression in epithelial tissues is directly driven by the transcription factor p63, which binds a p63-specific response element (RRRCGTGYYY) in the EVPL promoter that is distinct from the canonical p53 element; EVPL protein is abundant in skin and pharyngeal epithelium of wild-type mice but absent in p63-null mice [#1]. This p63-driven transcription is potentiated by BRD4, which physically interacts with p63 and co-occupies the EVPL locus to coordinate EVPL expression during keratinocyte differentiation [#6]. Beyond its structural role, EVPL acts as a tumor suppressor that restrains the RAS/ERK signaling axis: EVPL overexpression suppresses melanoma cell proliferation, migration, and invasion, promotes apoptosis, and dampens chemokine-driven macrophage recruitment and M2 polarization, with epistasis experiments placing EVPL upstream of ERK [#7]. In breast cancer cells EVPL is repressed by AGTPBP1, and restoring EVPL attenuates AGTPBP1-induced ERK activation and malignancy, defining an AGTPBP1/EVPL/ERK axis [#8]. Although EVPL maps within the tylosis esophageal cancer locus on 17q25, mutation and haplotype analyses excluded it as the causative TOC gene and found only rare somatic mutation in sporadic esophageal carcinoma [#2, #3].\",\n  \"teleology\": [\n    {\n      \"year\": 1996,\n      \"claim\": \"Established the molecular identity and candidate function of EVPL by placing it in the plakin family and mapping its locus, framing it as a structural connector of desmosomes, keratin filaments, and the cornified envelope.\",\n      \"evidence\": \"Somatic cell hybrid analysis and in situ hybridisation with cosmid/YAC isolation, plus homology to desmoplakin/BPAG1/plectin\",\n      \"pmids\": [\"8938451\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural linker role inferred from homology and expression, not directly tested\", \"No direct demonstration of binding to desmosomal or keratin components\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Tested whether EVPL is the causative gene at the tylosis esophageal cancer (TOC) locus and excluded it, redirecting the search for the TOC gene.\",\n      \"evidence\": \"Mutation and haplotype analysis with radiation hybrid and physical mapping across three TOC families\",\n      \"pmids\": [\"10409435\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Negative result does not address whether EVPL contributes to esophageal pathology by other mechanisms\", \"Limited to three families\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Resolved how EVPL is transcriptionally controlled, showing it is a direct p63 target via a p63-discriminating promoter element, linking EVPL to epithelial differentiation programs.\",\n      \"evidence\": \"Microarray, mutagenized-promoter luciferase assays, EMSA, and p63 knockout mouse immunostaining\",\n      \"pmids\": [\"15988020\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not establish EVPL's downstream functional consequences\", \"Cofactors of p63 at the EVPL promoter not defined in this study\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Assessed whether EVPL is somatically targeted in sporadic esophageal squamous cell carcinoma and found mutation is rare despite frequent LOH, arguing against EVPL as the primary driver at the locus.\",\n      \"evidence\": \"RT-PCR SSCP and direct sequencing of EVPL in OSC cell lines and tumors\",\n      \"pmids\": [\"15756445\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single missense variant of unknown functional impact\", \"Does not rule out epigenetic or expression-level inactivation\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Mapped the autoantigenic regions of EVPL in paraneoplastic pemphigus, localizing dominant epitopes to defined N-terminal and linker subdomains.\",\n      \"evidence\": \"ELISA of patient/control sera against 12 overlapping recombinant EVPL fragments\",\n      \"pmids\": [\"27427435\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not address whether autoantibodies are pathogenic\", \"Epitope mapping does not bear on normal EVPL function\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Linked EVPL loss to therapy resistance, showing EVPL knockdown increases radioresistance in esophageal cancer cells with inferred suppression of inflammatory signaling.\",\n      \"evidence\": \"RNAi knockdown in KYSE-150 with clonogenic survival assay and RNA-seq\",\n      \"pmids\": [\"33633453\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"NF-\\u03baB/TNF pathway involvement inferred from transcriptome, not directly validated\", \"Single cell line, single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified BRD4 as a transcriptional coactivator partnering with p63 to drive EVPL, refining the regulatory logic of EVPL expression during keratinocyte differentiation.\",\n      \"evidence\": \"Co-IP, ChIP, RT-qPCR, transcriptomics, and JQ1/knockdown perturbation\",\n      \"pmids\": [\"39605045\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"EVPL is one of several co-regulated targets; EVPL-specific dependence on BRD4 not isolated\", \"Direct vs indirect BRD4 effect at EVPL not fully separated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined a functional tumor-suppressor role for EVPL upstream of ERK, showing EVPL restrains melanoma proliferation, invasion, and macrophage recruitment through RAS/ERK signaling.\",\n      \"evidence\": \"Gain/loss-of-function in melanoma cells with functional assays, co-culture, and ERK-inhibitor epistasis (SCH772984)\",\n      \"pmids\": [\"38504693\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism by which EVPL regulates RAS/ERK is undefined\", \"Single lab, in vitro context\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Placed EVPL within a defined regulatory axis in breast cancer, showing AGTPBP1 represses EVPL to activate ERK and that restoring EVPL reverses malignancy.\",\n      \"evidence\": \"RNA-seq, gain/loss-of-function, and rescue experiments with pERK1/2 readout in two cell lines\",\n      \"pmids\": [\"41905574\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of AGTPBP1-mediated EVPL repression not established\", \"Direct biochemical link between EVPL and ERK pathway components missing\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The molecular mechanism by which EVPL, a structural cornified-envelope protein, suppresses RAS/ERK signaling remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No identified physical link between EVPL and RAS/ERK pathway components\", \"Whether the tumor-suppressive activity depends on EVPL's structural/desmosomal role is unknown\", \"No in vivo validation of the tumor-suppressor function\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7, 8]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [1, 6]}\n    ],\n    \"complexes\": [],\n    \"partners\": [],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":6,"faith_pct":83.33333333333333}}