{"gene":"EPCAM","run_date":"2026-04-28T17:46:03","timeline":{"discoveries":[{"year":2009,"finding":"EpCAM undergoes regulated intramembrane proteolysis (RIP): TACE sequentially sheds the ectodomain (EpEX) and presenilin-2 cleaves the remaining stub, releasing the intracellular domain (EpICD). EpICD then associates with FHL2, β-catenin, and Lef-1 to form a nuclear complex that contacts DNA at Lef-1 consensus sites, induces target gene transcription (including c-myc), and is oncogenic in immunodeficient mice.","method":"Pharmacological inhibition and genetic silencing of TACE and presenilin-2; nuclear fractionation; Co-IP of EpICD/FHL2/β-catenin/Lef-1 complex; reporter assays; xenograft tumor formation","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods (inhibitor + siRNA + co-IP + reporter + in vivo), strong mechanistic resolution, high citation count","pmids":["19136966"],"is_preprint":false},{"year":2004,"finding":"The intracellular domain of EpCAM is necessary and sufficient to upregulate c-myc and cyclins A/E, reduce growth-factor dependence, and enhance colony formation, establishing a direct link between EpCAM intracellular signaling and cell-cycle progression.","method":"Domain-swapping experiments; antisense-mediated knockdown; metabolic activity and colony formation assays in human 293 cells and NIH3T3 fibroblasts","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — domain-swap mutagenesis with loss-of-function and gain-of-function, replicated in two cell systems","pmids":["15195135"],"is_preprint":false},{"year":2004,"finding":"Silencing EpCAM with siRNA decreases breast cancer cell proliferation, migration, and invasion, and increases the detergent-insoluble (membrane-associated) fractions of E-cadherin, α-catenin, and β-catenin, indicating EpCAM negatively regulates E-cadherin-mediated adhesion complexes.","method":"siRNA knockdown; proliferation, migration, and invasion assays; detergent-solubility fractionation of cadherin/catenin complexes","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — clean KD with multiple phenotypic readouts in four cell lines, single lab","pmids":["15313925"],"is_preprint":false},{"year":2009,"finding":"Initial activation of EpCAM cleavage requires cell-to-cell contact (juxtacrine signaling); once EpEX is released it can act in a paracrine manner. EpICD-driven nuclear translocation and c-myc induction require the cleavage step and subsequent nuclear import.","method":"Confocal microscopy; immunoblotting; conditional cell-density experiments; conditional EpICD expression system","journal":"BMC cancer","confidence":"Medium","confidence_rationale":"Tier 2 — mechanistic dissection with cell-contact manipulation and conditional expression, single lab","pmids":["19925656"],"is_preprint":false},{"year":2012,"finding":"EpCAM promotes cell-cycle progression by transcriptionally upregulating cyclin D1 in a manner dependent on its direct interaction partner FHL2; downstream consequences include Rb phosphorylation and induction of cyclins E and A.","method":"EpCAM overexpression and knockdown; cyclin D1 promoter assays; co-IP of EpICD–FHL2; immunohistochemistry for Ki67, cyclin D1, and phospho-Rb in patient tumors","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — reporter assay + co-IP + in vivo correlation, moderate evidence from single lab with orthogonal methods","pmids":["22391566"],"is_preprint":false},{"year":2013,"finding":"EpCAM physically associates with claudin-7 (and claudin-1 via claudin-7) and protects them from lysosomal degradation; EpCAM knockdown reduces claudin-7 and claudin-1 protein levels and paradoxically increases their accumulation at tight junctions, altering trans-epithelial resistance.","method":"Preparative immunoprecipitation; co-IP; shRNA knockdown; trans-epithelial electroresistance measurements; immunofluorescence microscopy with lysosome inhibitors","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — reciprocal co-IP + functional rescue with lysosome inhibitors + shRNA phenotype, strong evidence from multiple orthogonal approaches","pmids":["23486470"],"is_preprint":false},{"year":2009,"finding":"EpCAM–claudin-7 interaction requires an AxxxG motif in EpCAM's transmembrane region; the resulting complex is recruited into tetraspanin-enriched membrane microdomains (TEMs) where it supports sustained ERK1/2 phosphorylation, upregulation of anti-apoptotic proteins, drug resistance, and enhanced cell motility, but abolishes EpCAM-mediated homophilic adhesion.","method":"Transfection of deletion and point-mutant EpCAM/claudin-7 constructs in HEK293 and BSp73AS cells; co-IP; TEM fractionation; proliferation, migration, and tumorigenicity assays","journal":"Molecular cancer research","confidence":"High","confidence_rationale":"Tier 1–2 — mutagenesis identifying specific transmembrane motif + co-IP + TEM fractionation + functional assays","pmids":["19276185"],"is_preprint":false},{"year":2013,"finding":"EpCAM acts as a potent inhibitor of novel PKC (nPKC) via a short pseudosubstrate-like segment in its cytoplasmic tail that binds nPKCs with high affinity; loss of EpCAM in Xenopus embryos leads to PKC overstimulation, ERK pathway hyperactivation, exacerbated myosin contractility, loss of cadherin-mediated adhesion, and tissue dissociation.","method":"Xenopus embryo loss-of-function; in vitro PKC binding assay with cytoplasmic tail peptides; mutagenesis; myosin contractility measurements; cadherin adhesion assays","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 1 — in vitro binding assay with mutagenesis + in vivo phenotype in Xenopus, strong mechanistic resolution","pmids":["24183651"],"is_preprint":false},{"year":2017,"finding":"Matriptase cleaves EpCAM after Arg80; loss of the matriptase inhibitor HAI-2 in intestinal epithelial cells leads to unrestrained matriptase activity, efficient EpCAM cleavage, reduced EpCAM–claudin-7 association, and lysosomal degradation of both EpCAM and claudin-7, causing intestinal epithelial dysplasia (congenital tufting enteropathy phenotype).","method":"In vitro cleavage of purified recombinant proteins; cell transfection; HAI-2 knockout mouse model; immunoprecipitation; HAI-2 CTE mutant functional rescue experiments","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution + mutagenesis + knockout mouse model with defined disease phenotype, strong mechanistic resolution","pmids":["28094766"],"is_preprint":false},{"year":2018,"finding":"EpCAM homo-oligomerization does not mediate intercellular adhesion; EpCAM monomers do not form inter-cellular oligomers detectable by SAXS or XL-MS, and bead aggregation assays confirm no homophilic adhesion. EpCAM forms stable cis-dimers on the cell surface at pre-formed cell–cell contacts as detected by FLIM-FRET.","method":"SAXS; cross-linking mass spectrometry (XL-MS); bead aggregation assay; FLIM-FRET on live cells","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1 — multiple biophysical methods (SAXS, XL-MS, FLIM-FRET, bead aggregation) in one study","pmids":["30185875"],"is_preprint":false},{"year":2018,"finding":"The extracellular domain of EpCAM (EpEX) activates EGFR and downstream ERK1/2 signaling to promote colon cancer cell migration and proliferation; EpEX–EGFR–ERK1/2 signaling positively regulates RIP of EpCAM and EpICD shedding, which then drives nuclear β-catenin accumulation and HIF1α target gene expression.","method":"EGFR/MEK inhibitor treatment; EpEX-stimulation assays; immunoprecipitation; Western blotting; xenograft tumor models; analysis of colorectal cancer tissues","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological inhibition + IP + in vivo validation, single lab with multiple orthogonal approaches","pmids":["29981429"],"is_preprint":false},{"year":2011,"finding":"EpCAM and its complex partner claudin-7 (Cldn7) facilitate transcription-factor-mediated somatic cell reprogramming to iPSCs; overexpression of EpCAM or EpICD enhances reprogramming efficiency, activates the Oct4 promoter, and suppresses p53 and p21 expression.","method":"Quantitative RT-PCR; alkaline phosphatase and Nanog colony assays; EpCAM/EpICD overexpression; shRNA knockdown; Oct4 promoter reporter assay; iPSC characterization in vitro and in vivo","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — gain/loss-of-function with promoter reporter and iPSC functional characterization, single lab","pmids":["21799003"],"is_preprint":false},{"year":2008,"finding":"Mutations in EPCAM (including a splice-site mutation causing deletion of exon 4) are the genetic cause of congenital tufting enteropathy, resulting in loss of EpCAM protein from intestinal epithelium.","method":"SNP homozygosity mapping; direct DNA sequencing; RT-PCR; immunohistochemistry; Western blotting of patient intestinal tissue","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 2 — genetic cause established by sequencing + protein confirmation in patient tissue, replicated in multiple unrelated patients","pmids":["18572020"],"is_preprint":false},{"year":2011,"finding":"3′-end deletions of EPCAM encompassing the transcription termination signal cause allele-specific epigenetic silencing (promoter hypermethylation) of the neighboring DNA mismatch repair gene MSH2 in tissues expressing EPCAM, leading to Lynch syndrome.","method":"Multiplex ligation-dependent probe amplification (MLPA); MSH2 promoter methylation analysis; tissue-specific expression profiling in germline deletion carriers","journal":"Human mutation","confidence":"High","confidence_rationale":"Tier 2 — mechanism (read-through transcription silencing MSH2) confirmed in 45 families with methylation analysis, strong replicated evidence","pmids":["21309036"],"is_preprint":false},{"year":2013,"finding":"EpCAM mutation (exon 4 deletion) causes loss of EpCAM–claudin-7 colocalization and complex formation in vivo, leading to intestinal barrier dysfunction (increased permeability, reduced ion transport, altered desmosomes) and the CTE pathological phenotype.","method":"Cre-LoxP conditional mouse model; histology; electron microscopy; immunohistochemistry; barrier permeability assays; ion transport measurements; comparison with CTE patient tissue","journal":"American journal of physiology. Gastrointestinal and liver physiology","confidence":"High","confidence_rationale":"Tier 2 — in vivo knockout mouse model with defined molecular (EpCAM/claudin-7 complex disruption) and functional (barrier/transport) phenotype, validated in human CTE tissue","pmids":["24337010"],"is_preprint":false},{"year":2011,"finding":"EpCAM signaling activates NF-κB, promotes c-Jun phosphorylation via the JNK pathway, and increases AP-1 transcription factor activity; soluble extracellular EpCAM (rEpEX) can rescue invasion, AP-1 activity, and c-Jun phosphorylation after EpCAM ablation.","method":"RNAi loss-of-function; gain-of-function with cDNA construct; recombinant EpEX rescue; phosphoprotein analysis; AP-1 and JNK reporter/inhibitor experiments; invasion assays in vitro and in vivo","journal":"Breast cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — RNAi + constitutively active constructs + pharmacological inhibitors + rescue experiments, single lab","pmids":["22132731"],"is_preprint":false},{"year":2013,"finding":"EpCAM ablation decreases NF-κB transcription factor activity and RELA phosphorylation, increases IκBα protein, and reduces IL-8 expression; forced expression of IκBα or RELA ablation blocks EpCAM-dependent rescue of IL-8 promoter activity.","method":"RNAi; constitutively active/dominant negative NF-κB constructs; IL-8 promoter reporter assay; ELISA; Western blotting","journal":"Molecular cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — reporter assay + genetic rescue experiments, single lab with multiple orthogonal approaches","pmids":["23378578"],"is_preprint":false},{"year":2019,"finding":"EpCAM associates with integrin β1 (shown by co-immunoprecipitation); EpCAM knockout reduces integrin α5 expression and decreases phosphorylation of FAK, AKT, and ERK, impairing cell adhesion, migration, and proliferation on extracellular matrix proteins.","method":"CRISPR/Cas9 EpCAM knockout; co-immunoprecipitation; Western blotting for FAK/AKT/ERK phosphorylation; adhesion and migration assays","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 — CRISPR KO with co-IP and signaling readouts, single lab","pmids":["31806375"],"is_preprint":false},{"year":2020,"finding":"EpCAM organizes into heterogeneous clusters on the cell membrane and localizes within tetraspanin-enriched microdomains (TEMs) through co-association with CD9; cytoskeleton integrity and N-glycosylation both limit EpCAM cluster assembly.","method":"Direct stochastic optical reconstruction microscopy (dSTORM) with fluorophore-conjugated peptides; dual-color super-resolution imaging; CD9 knockdown; cytoskeleton disruption; glycosylation inhibition","journal":"Analytical chemistry","confidence":"Medium","confidence_rationale":"Tier 1–2 — super-resolution imaging with single-molecule precision + genetic/pharmacological perturbations, single lab","pmids":["31876148"],"is_preprint":false},{"year":2020,"finding":"Matriptase cleaves both EpCAM and its homolog TROP2 in keratinocytes; EpCAM and TROP2 are redundant in stabilizing claudin-1 and claudin-7 (knockdown of either alone has small effects, but combined knockdown markedly reduces claudins); HAI-1 (not HAI-2) is the principal physiological inhibitor of matriptase in keratinocytes.","method":"In vitro cleavage of purified recombinant proteins; siRNA knockdown of EpCAM, TROP2, HAI-1, HAI-2, matriptase in HaCaT cells; Western blotting; lysosome inhibitor (chloroquine) rescue","journal":"Cells","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution with purified proteins + cell-based genetic validation with multiple orthogonal siRNA knockdowns","pmids":["32326212"],"is_preprint":false},{"year":2023,"finding":"The extracellular domain of EpCAM (EpEX) binds directly to HGFR (c-Met) and cooperates with HGF to activate downstream ERK and FAK-AKT signaling; EpEX stabilizes β-catenin and Snail by decreasing GSK3β activity, promoting EMT and metastasis in colon cancer.","method":"Immunoprecipitation; ELISA; FRET; Western blotting; migration/invasion assays; tail vein injection and orthotopic metastasis mouse models","journal":"Journal of translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP/FRET demonstrating direct EpEX–HGFR interaction + in vivo metastasis model, single lab","pmids":["37543570"],"is_preprint":false}],"current_model":"EpCAM is a multifunctional transmembrane glycoprotein that undergoes regulated intramembrane proteolysis (by TACE and presenilin-2, initiated by cell-to-cell contact) to release a soluble ectodomain (EpEX) and a nuclear intracellular domain (EpICD); EpICD forms a complex with FHL2, β-catenin, and Lef-1 to drive transcription of proliferative target genes (c-myc, cyclin D1); EpEX acts as a ligand for EGFR and HGFR to activate ERK, AKT, and downstream pathways; EpCAM also directly inhibits novel PKC via a pseudosubstrate motif in its cytoplasmic tail to suppress actomyosin contractility, interacts with claudin-7 and claudin-1 to stabilize them from lysosomal degradation (a function regulated by the matriptase/HAI protease axis), and associates with integrin β1 to modulate FAK/ERK signaling, while germline mutations cause congenital tufting enteropathy and 3′ deletions silence MSH2 to cause Lynch syndrome."},"narrative":{"teleology":[{"year":2004,"claim":"Establishing that EpCAM is not merely an adhesion molecule but an active signaling receptor whose intracellular domain is necessary and sufficient for upregulating c-myc and cyclins and promoting proliferation answered the fundamental question of whether EpCAM has cell-autonomous oncogenic signaling capacity.","evidence":"Domain-swap mutagenesis with gain/loss-of-function in HEK293 and NIH3T3 cells; siRNA knockdown in breast cancer lines with proliferation, migration, and cadherin fractionation readouts","pmids":["15195135","15313925"],"confidence":"High","gaps":["Protease responsible for EpICD release unknown","Identity of nuclear partners of EpICD not yet defined","Mechanism linking EpCAM to E-cadherin regulation unclear"]},{"year":2008,"claim":"Identification of homozygous EPCAM mutations as the genetic cause of congenital tufting enteropathy established EpCAM as essential for intestinal epithelial integrity and linked it to a Mendelian disease.","evidence":"SNP homozygosity mapping and sequencing in multiple unrelated CTE families; immunohistochemistry confirming protein loss in patient intestinal tissue","pmids":["18572020"],"confidence":"High","gaps":["Molecular mechanism of epithelial disruption upon EpCAM loss not defined","Claudin interactions not yet identified"]},{"year":2009,"claim":"Discovery of the complete regulated intramembrane proteolysis (RIP) cascade — TACE ectodomain shedding followed by presenilin-2 cleavage releasing EpICD, which assembles a nuclear FHL2/β-catenin/Lef-1 transcription complex — provided the full mechanistic pathway from membrane to gene activation.","evidence":"Pharmacological and siRNA inhibition of TACE and presenilin-2; nuclear fractionation and co-IP of EpICD complex; reporter assays; xenograft tumorigenesis; cell-contact density experiments showing juxtacrine initiation","pmids":["19136966","19925656"],"confidence":"High","gaps":["Crystal structure of EpICD-containing nuclear complex unavailable","Upstream signals triggering TACE activation at cell contacts not defined"]},{"year":2009,"claim":"Mapping the EpCAM–claudin-7 interaction to a transmembrane AxxxG motif and demonstrating that this complex is recruited to tetraspanin-enriched microdomains (TEMs) where it sustains ERK1/2 signaling and drug resistance revealed a second, adhesion-independent oncogenic function of EpCAM.","evidence":"Deletion and point-mutant constructs in HEK293/BSp73AS cells; co-IP; TEM fractionation; proliferation, migration, and tumorigenicity assays","pmids":["19276185"],"confidence":"High","gaps":["Structural basis of the AxxxG-mediated transmembrane interaction not resolved","Relative contributions of RIP signaling versus TEM-based signaling to tumorigenesis unclear"]},{"year":2011,"claim":"Demonstration that 3′ EPCAM deletions cause read-through transcription and epigenetic silencing of MSH2 defined a cis-regulatory mechanism for Lynch syndrome independent of MSH2 coding mutations.","evidence":"MLPA and MSH2 promoter methylation analysis in 45 carrier families; tissue-specific expression profiling","pmids":["21309036"],"confidence":"High","gaps":["Precise chromatin mechanism of read-through-induced silencing not fully dissected","Genotype–phenotype correlation for different deletion sizes incomplete"]},{"year":2012,"claim":"Identification of cyclin D1 as a direct transcriptional target of EpICD–FHL2 signaling, with consequent Rb phosphorylation, connected EpCAM proteolytic signaling to a canonical cell-cycle progression mechanism.","evidence":"EpCAM overexpression/knockdown; cyclin D1 promoter reporter; co-IP of EpICD–FHL2; Ki67 and cyclin D1 IHC in patient tumors","pmids":["22391566"],"confidence":"High","gaps":["Full set of EpICD transcriptional targets undefined","Chromatin occupancy by the EpICD complex not mapped genome-wide"]},{"year":2013,"claim":"Discovery that EpCAM's cytoplasmic tail contains a pseudosubstrate motif that directly inhibits novel PKC isoforms, thereby restraining myosin contractility and preserving cadherin adhesion, established a proteolysis-independent signaling function for full-length EpCAM.","evidence":"In vitro PKC binding assay with synthetic cytoplasmic-tail peptides; mutagenesis; Xenopus embryo loss-of-function with actomyosin and cadherin phenotyping","pmids":["24183651"],"confidence":"High","gaps":["Whether nPKC inhibition is operative in mammalian epithelia not confirmed","Relative importance of PKC-inhibitory versus RIP functions in CTE pathogenesis unknown"]},{"year":2013,"claim":"Showing that EpCAM protects claudin-7 and claudin-1 from lysosomal degradation, and that CTE-causing EpCAM mutations ablate EpCAM–claudin-7 complex formation causing intestinal barrier dysfunction, provided a molecular explanation for congenital tufting enteropathy.","evidence":"Reciprocal co-IP; shRNA knockdown with lysosome inhibitor rescue; conditional EpCAM-knockout mouse; electron microscopy; barrier permeability and ion transport assays; comparison with CTE patient tissue","pmids":["23486470","24337010"],"confidence":"High","gaps":["Whether claudin destabilization alone accounts for the full CTE phenotype is untested","Role of desmosome alterations observed in KO mice not mechanistically dissected"]},{"year":2017,"claim":"Identification of matriptase as the protease cleaving EpCAM at Arg80, with HAI-2 as its inhibitor, showed that uncontrolled matriptase activity phenocopies CTE by destabilizing the EpCAM–claudin-7 complex via ectodomain cleavage.","evidence":"In vitro cleavage with purified proteins; HAI-2 knockout mouse; co-IP; CTE mutant rescue","pmids":["28094766"],"confidence":"High","gaps":["Whether matriptase cleavage feeds into RIP-dependent EpICD signaling or only triggers degradation is unclear","Tissue-specific regulation of matriptase/HAI balance not mapped comprehensively"]},{"year":2018,"claim":"Biophysical evidence that EpCAM does not mediate intercellular homophilic adhesion but forms cis-dimers at pre-existing contacts overturned the long-held model of EpCAM as a cell-adhesion molecule and reframed it primarily as a signaling receptor.","evidence":"SAXS, cross-linking mass spectrometry, bead aggregation assay, and FLIM-FRET on live cells","pmids":["30185875"],"confidence":"High","gaps":["Functional role of cis-dimerization not established","Whether cis-dimers are the signaling-competent unit for RIP is untested"]},{"year":2018,"claim":"Showing that the shed ectodomain EpEX activates EGFR–ERK1/2 signaling to promote proliferation and migration, and that this pathway feeds back to stimulate further EpCAM RIP and nuclear β-catenin accumulation, established a positive-feedback loop linking extracellular and intracellular EpCAM signals.","evidence":"EGFR/MEK inhibitor treatment; EpEX stimulation; co-IP; xenograft models; colorectal cancer tissue analysis","pmids":["29981429"],"confidence":"Medium","gaps":["Direct EpEX–EGFR binding site not mapped","Relative importance of autocrine versus paracrine EpEX signaling in vivo unknown"]},{"year":2020,"claim":"Super-resolution imaging revealed that EpCAM forms heterogeneous nanoclusters within CD9-positive tetraspanin microdomains, regulated by cytoskeletal integrity and N-glycosylation, providing a spatial framework for its signaling functions. Parallel work confirmed functional redundancy between EpCAM and TROP2 in stabilizing claudins against matriptase-mediated degradation.","evidence":"dSTORM with CD9 knockdown and pharmacological perturbations; in vitro cleavage assays with siRNA knockdown of EpCAM/TROP2/matriptase/HAI-1/HAI-2 in keratinocytes","pmids":["31876148","32326212"],"confidence":"Medium","gaps":["How nanocluster organization relates to RIP activation is unknown","TROP2 redundancy complicates tissue-specific phenotype predictions"]},{"year":2023,"claim":"Demonstration that EpEX directly binds HGFR (c-Met) and cooperates with HGF to activate ERK and FAK-AKT signaling, stabilize β-catenin/Snail, and promote EMT and metastasis expanded the receptor repertoire of shed EpEX beyond EGFR.","evidence":"Co-IP, FRET, ELISA for direct EpEX–HGFR interaction; migration/invasion assays; orthotopic and tail-vein metastasis mouse models","pmids":["37543570"],"confidence":"Medium","gaps":["EpEX binding site on HGFR not mapped structurally","Whether EpEX activates additional RTKs is unknown","Relative contribution of EGFR versus HGFR arm in vivo not delineated"]},{"year":null,"claim":"Key unresolved questions include how cis-dimerization and nanocluster organization couple to RIP initiation, the genome-wide transcriptional program of the EpICD nuclear complex, and how the PKC-inhibitory and claudin-stabilizing functions of full-length EpCAM are coordinated with proteolytic signaling in different epithelial contexts.","evidence":"","pmids":[],"confidence":"Low","gaps":["No ChIP-seq or CUT&RUN mapping of EpICD-containing complex","Structural basis of EpICD–FHL2–β-catenin–Lef-1 assembly unknown","Relative contribution of PKC inhibition versus RIP signaling to epithelial homeostasis not dissected in mammalian models"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[7]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[0,1,4]},{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[10,20]},{"term_id":"GO:0098631","term_label":"cell adhesion mediator activity","supporting_discovery_ids":[5,14]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6,9,18]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,3,4]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,7,10,15,17,20]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[1,4]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0,4,16]},{"term_id":"R-HSA-1500931","term_label":"Cell-Cell communication","supporting_discovery_ids":[5,14]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[12,13]}],"complexes":["EpICD/FHL2/β-catenin/Lef-1 nuclear transcription complex","EpCAM–claudin-7 membrane complex","Tetraspanin-enriched microdomain (TEM) complex"],"partners":["FHL2","CTNNB1","CLDN7","CLDN1","EGFR","MET","ITGB1","CD9"],"other_free_text":[]},"mechanistic_narrative":"EpCAM is a transmembrane glycoprotein that functions as a signaling hub in epithelial proliferation, adhesion, and differentiation through regulated intramembrane proteolysis (RIP) and direct modulation of multiple signaling pathways. TACE-mediated ectodomain shedding followed by presenilin-2 cleavage releases a soluble ectodomain (EpEX) that activates EGFR and HGFR to stimulate ERK, AKT, and FAK signaling, and an intracellular domain (EpICD) that forms a nuclear complex with FHL2, β-catenin, and Lef-1 to drive transcription of c-myc and cyclin D1, promoting cell-cycle progression [PMID:19136966, PMID:15195135, PMID:22391566, PMID:29981429, PMID:37543570]. EpCAM directly inhibits novel PKC isoforms via a pseudosubstrate motif in its cytoplasmic tail, thereby restraining actomyosin contractility and maintaining cadherin-mediated adhesion, and it physically associates with claudin-7 through a transmembrane AxxxG motif to protect claudins from matriptase-initiated lysosomal degradation — a mechanism whose disruption causes congenital tufting enteropathy [PMID:24183651, PMID:23486470, PMID:19276185, PMID:28094766, PMID:18572020]. Germline 3′ deletions of EPCAM additionally cause epigenetic silencing of the adjacent MSH2 gene, resulting in Lynch syndrome [PMID:21309036]."},"prefetch_data":{"uniprot":{"accession":"P16422","full_name":"Epithelial cell adhesion molecule","aliases":["Adenocarcinoma-associated antigen","Cell surface glycoprotein Trop-1","Epithelial cell surface antigen","Epithelial glycoprotein","EGP","Epithelial glycoprotein 314","EGP314","hEGP314","KS 1/4 antigen","KSA","Major gastrointestinal tumor-associated protein GA733-2","Tumor-associated calcium signal transducer 1"],"length_aa":314,"mass_kda":34.9,"function":"May act as a physical homophilic interaction molecule between intestinal epithelial cells (IECs) and intraepithelial lymphocytes (IELs) at the mucosal epithelium for providing immunological barrier as a first line of defense against mucosal infection. Plays a role in embryonic stem cells proliferation and differentiation. Up-regulates the expression of FABP5, MYC and cyclins A and E","subcellular_location":"Lateral cell membrane; Cell junction, tight junction","url":"https://www.uniprot.org/uniprotkb/P16422/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EPCAM","classification":"Not Classified","n_dependent_lines":21,"n_total_lines":1208,"dependency_fraction":0.0173841059602649},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/EPCAM","total_profiled":1310},"omim":[{"mim_id":"620517","title":"STERILE ALPHA MOTIF DOMAIN-CONTAINING PROTEIN 5; SAMD5","url":"https://www.omim.org/entry/620517"},{"mim_id":"613244","title":"LYNCH SYNDROME 8; LYNCH8","url":"https://www.omim.org/entry/613244"},{"mim_id":"613217","title":"DIARRHEA 5, WITH TUFTING ENTEROPATHY, CONGENITAL; DIAR5","url":"https://www.omim.org/entry/613217"},{"mim_id":"609309","title":"MutS HOMOLOG 2; MSH2","url":"https://www.omim.org/entry/609309"},{"mim_id":"608576","title":"GRAINYHEAD-LIKE 2; GRHL2","url":"https://www.omim.org/entry/608576"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Plasma membrane","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"intestine","ntpm":756.2}],"url":"https://www.proteinatlas.org/search/EPCAM"},"hgnc":{"alias_symbol":["Ly74","TROP1","GA733-2","EGP34","EGP40","EGP-2","KSA","CD326","Ep-CAM","HEA125","KS1/4","MK-1","MH99","MOC31","MOC-31","323/A3","17-1A","TACST-1","CO-17A","ESA","BerEp4","Ber-Ep4"],"prev_symbol":["M4S1","MIC18","TACSTD1"]},"alphafold":{"accession":"P16422","domains":[{"cath_id":"-","chopping":"36-76_92-254","consensus_level":"medium","plddt":93.7632,"start":36,"end":254}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P16422","model_url":"https://alphafold.ebi.ac.uk/files/AF-P16422-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P16422-F1-predicted_aligned_error_v6.png","plddt_mean":87.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EPCAM","jax_strain_url":"https://www.jax.org/strain/search?query=EPCAM"},"sequence":{"accession":"P16422","fasta_url":"https://rest.uniprot.org/uniprotkb/P16422.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P16422/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P16422"}},"corpus_meta":[{"pmid":"19136966","id":"PMC_19136966","title":"Nuclear signalling by tumour-associated antigen EpCAM.","date":"2009","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/19136966","citation_count":564,"is_preprint":false},{"pmid":"15313925","id":"PMC_15313925","title":"EpCAM is overexpressed in breast cancer and is a potential target for breast cancer gene therapy.","date":"2004","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/15313925","citation_count":442,"is_preprint":false},{"pmid":"19584271","id":"PMC_19584271","title":"The emerging role of EpCAM in cancer and stem cell signaling.","date":"2009","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/19584271","citation_count":433,"is_preprint":false},{"pmid":"15195135","id":"PMC_15195135","title":"The carcinoma-associated antigen EpCAM upregulates c-myc and induces cell proliferation.","date":"2004","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/15195135","citation_count":296,"is_preprint":false},{"pmid":"32507912","id":"PMC_32507912","title":"Expression and function of epithelial cell adhesion molecule EpCAM: where are we after 40 years?","date":"2020","source":"Cancer metastasis reviews","url":"https://pubmed.ncbi.nlm.nih.gov/32507912","citation_count":258,"is_preprint":false},{"pmid":"20837599","id":"PMC_20837599","title":"EpCAM in carcinogenesis: the good, the bad or the ugly.","date":"2010","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/20837599","citation_count":255,"is_preprint":false},{"pmid":"21415054","id":"PMC_21415054","title":"EpCAM expression in primary tumour tissues and metastases: an immunohistochemical analysis.","date":"2011","source":"Journal of clinical pathology","url":"https://pubmed.ncbi.nlm.nih.gov/21415054","citation_count":231,"is_preprint":false},{"pmid":"23618806","id":"PMC_23618806","title":"EpCAM: structure and function in health and disease.","date":"2013","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/23618806","citation_count":227,"is_preprint":false},{"pmid":"26184843","id":"PMC_26184843","title":"The detection of EpCAM(+) and EpCAM(-) circulating tumor cells.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26184843","citation_count":223,"is_preprint":false},{"pmid":"2333300","id":"PMC_2333300","title":"Molecular cloning of cDNA for the carcinoma-associated antigen GA733-2.","date":"1990","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/2333300","citation_count":165,"is_preprint":false},{"pmid":"18572020","id":"PMC_18572020","title":"Identification of EpCAM as the gene for congenital tufting enteropathy.","date":"2008","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/18572020","citation_count":157,"is_preprint":false},{"pmid":"19075676","id":"PMC_19075676","title":"CD44 and EpCAM: cancer-initiating cell markers.","date":"2008","source":"Current molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/19075676","citation_count":148,"is_preprint":false},{"pmid":"31225512","id":"PMC_31225512","title":"Biology and clinical relevance of EpCAM.","date":"2019","source":"Cell stress","url":"https://pubmed.ncbi.nlm.nih.gov/31225512","citation_count":140,"is_preprint":false},{"pmid":"30015855","id":"PMC_30015855","title":"Functions of EpCAM in physiological processes and diseases (Review).","date":"2018","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/30015855","citation_count":128,"is_preprint":false},{"pmid":"21309036","id":"PMC_21309036","title":"Recurrence and variability of germline EPCAM deletions in Lynch syndrome.","date":"2011","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/21309036","citation_count":123,"is_preprint":false},{"pmid":"23486470","id":"PMC_23486470","title":"Epithelial cell adhesion molecule (EpCAM) regulates claudin dynamics and tight junctions.","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23486470","citation_count":112,"is_preprint":false},{"pmid":"22391566","id":"PMC_22391566","title":"EpCAM regulates cell cycle progression via control of cyclin D1 expression.","date":"2012","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/22391566","citation_count":99,"is_preprint":false},{"pmid":"19276185","id":"PMC_19276185","title":"Claudin-7 regulates EpCAM-mediated functions in tumor progression.","date":"2009","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/19276185","citation_count":99,"is_preprint":false},{"pmid":"21224371","id":"PMC_21224371","title":"Circulating tumor cells and EpCAM expression in neuroendocrine tumors.","date":"2011","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/21224371","citation_count":90,"is_preprint":false},{"pmid":"29295696","id":"PMC_29295696","title":"Antibody Based EpCAM Targeted Therapy of Cancer, Review and Update.","date":"2018","source":"Current cancer drug targets","url":"https://pubmed.ncbi.nlm.nih.gov/29295696","citation_count":87,"is_preprint":false},{"pmid":"32046162","id":"PMC_32046162","title":"Revisiting the Roles of Pro-Metastatic EpCAM in Cancer.","date":"2020","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/32046162","citation_count":83,"is_preprint":false},{"pmid":"17325709","id":"PMC_17325709","title":"EpCAM an immunotherapeutic target for gastrointestinal malignancy: current experience and future challenges.","date":"2007","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/17325709","citation_count":80,"is_preprint":false},{"pmid":"15613858","id":"PMC_15613858","title":"Expression of epithelial cell adhesion molecule (EpCam) in renal epithelial tumors.","date":"2005","source":"The American journal of surgical pathology","url":"https://pubmed.ncbi.nlm.nih.gov/15613858","citation_count":79,"is_preprint":false},{"pmid":"21799003","id":"PMC_21799003","title":"Epithelial cell adhesion molecule (EpCAM) complex proteins promote transcription factor-mediated pluripotency reprogramming.","date":"2011","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21799003","citation_count":78,"is_preprint":false},{"pmid":"34209658","id":"PMC_34209658","title":"Functional Implications of the Dynamic Regulation of EpCAM during Epithelial-to-Mesenchymal Transition.","date":"2021","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/34209658","citation_count":74,"is_preprint":false},{"pmid":"24141784","id":"PMC_24141784","title":"Context-dependent adaption of EpCAM expression in early systemic esophageal cancer.","date":"2013","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/24141784","citation_count":73,"is_preprint":false},{"pmid":"36369033","id":"PMC_36369033","title":"Understanding the versatile roles and applications of EpCAM in cancers: from bench to bedside.","date":"2022","source":"Experimental hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/36369033","citation_count":72,"is_preprint":false},{"pmid":"20012351","id":"PMC_20012351","title":"Epithelial cell adhesion molecule (EpCAM) is overexpressed in breast cancer metastases.","date":"2009","source":"Breast cancer research and treatment","url":"https://pubmed.ncbi.nlm.nih.gov/20012351","citation_count":72,"is_preprint":false},{"pmid":"29158811","id":"PMC_29158811","title":"Transforming doxorubicin into a cancer stem cell killer via EpCAM aptamer-mediated delivery.","date":"2017","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/29158811","citation_count":71,"is_preprint":false},{"pmid":"29981429","id":"PMC_29981429","title":"Extracellular domain of EpCAM enhances tumor progression through EGFR signaling in colon cancer cells.","date":"2018","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/29981429","citation_count":69,"is_preprint":false},{"pmid":"26264278","id":"PMC_26264278","title":"Gene Knockdown by EpCAM Aptamer-siRNA Chimeras Suppresses Epithelial Breast Cancers and Their Tumor-Initiating Cells.","date":"2015","source":"Molecular cancer therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/26264278","citation_count":68,"is_preprint":false},{"pmid":"25576037","id":"PMC_25576037","title":"EpCAM aptamer mediated cancer cell specific delivery of EpCAM siRNA using polymeric nanocomplex.","date":"2015","source":"Journal of biomedical science","url":"https://pubmed.ncbi.nlm.nih.gov/25576037","citation_count":66,"is_preprint":false},{"pmid":"22718399","id":"PMC_22718399","title":"Role of the EpCAM (CD326) in prostate cancer metastasis and progression.","date":"2012","source":"Cancer metastasis reviews","url":"https://pubmed.ncbi.nlm.nih.gov/22718399","citation_count":65,"is_preprint":false},{"pmid":"19925656","id":"PMC_19925656","title":"Initial activation of EpCAM cleavage via cell-to-cell contact.","date":"2009","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/19925656","citation_count":65,"is_preprint":false},{"pmid":"28094766","id":"PMC_28094766","title":"Matriptase-mediated cleavage of EpCAM destabilizes claudins and dysregulates intestinal epithelial homeostasis.","date":"2017","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/28094766","citation_count":63,"is_preprint":false},{"pmid":"29627827","id":"PMC_29627827","title":"Lgr5+CD44+EpCAM+ Strictly Defines Cancer Stem Cells in Human Colorectal Cancer.","date":"2018","source":"Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/29627827","citation_count":59,"is_preprint":false},{"pmid":"30461124","id":"PMC_30461124","title":"EPCAM mutation update: Variants associated with congenital tufting enteropathy and Lynch syndrome.","date":"2018","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/30461124","citation_count":57,"is_preprint":false},{"pmid":"33627408","id":"PMC_33627408","title":"Immunotherapy for breast cancer using EpCAM aptamer tumor-targeted gene knockdown.","date":"2021","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/33627408","citation_count":55,"is_preprint":false},{"pmid":"10203061","id":"PMC_10203061","title":"Autoantibodies against the tumour-associated antigen GA733-2 in patients with colorectal carcinoma.","date":"1999","source":"Cancer immunology, immunotherapy : CII","url":"https://pubmed.ncbi.nlm.nih.gov/10203061","citation_count":51,"is_preprint":false},{"pmid":"31584203","id":"PMC_31584203","title":"Hypoxia modulates stem cell properties and induces EMT through N-glycosylation of EpCAM in breast cancer cells.","date":"2019","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/31584203","citation_count":50,"is_preprint":false},{"pmid":"24183651","id":"PMC_24183651","title":"EpCAM controls actomyosin contractility and cell adhesion by direct inhibition of PKC.","date":"2013","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/24183651","citation_count":48,"is_preprint":false},{"pmid":"32408542","id":"PMC_32408542","title":"A Novel Function for KLF4 in Modulating the De-differentiation of EpCAM-/CD133- nonStem Cells into EpCAM+/CD133+ Liver Cancer Stem Cells in HCC Cell Line HuH7.","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32408542","citation_count":48,"is_preprint":false},{"pmid":"24337010","id":"PMC_24337010","title":"Functional consequences of EpCam mutation in mice and men.","date":"2013","source":"American journal of physiology. Gastrointestinal and liver physiology","url":"https://pubmed.ncbi.nlm.nih.gov/24337010","citation_count":44,"is_preprint":false},{"pmid":"22388758","id":"PMC_22388758","title":"The molecular basis of EPCAM expression loss in Lynch syndrome-associated tumors.","date":"2012","source":"Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc","url":"https://pubmed.ncbi.nlm.nih.gov/22388758","citation_count":43,"is_preprint":false},{"pmid":"20426706","id":"PMC_20426706","title":"Adecatumumab: an anti-EpCAM monoclonal antibody, from the bench to the bedside.","date":"2010","source":"Expert opinion on biological therapy","url":"https://pubmed.ncbi.nlm.nih.gov/20426706","citation_count":41,"is_preprint":false},{"pmid":"26317650","id":"PMC_26317650","title":"An anti-EpCAM antibody EpAb2-6 for the treatment of colon cancer.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26317650","citation_count":41,"is_preprint":false},{"pmid":"25482158","id":"PMC_25482158","title":"Mutation of EpCAM leads to intestinal barrier and ion transport dysfunction.","date":"2014","source":"Journal of molecular medicine (Berlin, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/25482158","citation_count":40,"is_preprint":false},{"pmid":"22132731","id":"PMC_22132731","title":"Activator protein 1 (AP-1) contributes to EpCAM-dependent breast cancer invasion.","date":"2011","source":"Breast cancer research : BCR","url":"https://pubmed.ncbi.nlm.nih.gov/22132731","citation_count":40,"is_preprint":false},{"pmid":"30628064","id":"PMC_30628064","title":"Shedding light on the EpCAM: An overview.","date":"2019","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30628064","citation_count":39,"is_preprint":false},{"pmid":"21858196","id":"PMC_21858196","title":"Dynamic changes in EPCAM expression during spermatogonial stem cell differentiation in the mouse testis.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21858196","citation_count":39,"is_preprint":false},{"pmid":"32976980","id":"PMC_32976980","title":"EpCAM cellular functions in adhesion and migration, and potential impact on invasion: A critical review.","date":"2020","source":"Biochimica et biophysica acta. Reviews on cancer","url":"https://pubmed.ncbi.nlm.nih.gov/32976980","citation_count":37,"is_preprint":false},{"pmid":"25110875","id":"PMC_25110875","title":"Lynch-like syndrome: characterization and comparison with EPCAM deletion carriers.","date":"2014","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/25110875","citation_count":37,"is_preprint":false},{"pmid":"26493939","id":"PMC_26493939","title":"The role of EpCAM in physiology and pathology of the epithelium.","date":"2015","source":"Histology and histopathology","url":"https://pubmed.ncbi.nlm.nih.gov/26493939","citation_count":35,"is_preprint":false},{"pmid":"24727741","id":"PMC_24727741","title":"Coexpression of EpCAM, CD44 variant isoforms and claudin-7 in anaplastic thyroid carcinoma.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24727741","citation_count":35,"is_preprint":false},{"pmid":"24077349","id":"PMC_24077349","title":"Promoter hypomethylation of EpCAM-regulated bone morphogenetic protein gene family in recurrent endometrial cancer.","date":"2013","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/24077349","citation_count":35,"is_preprint":false},{"pmid":"26176230","id":"PMC_26176230","title":"EpCAM Aptamer-siRNA Chimera Targets and Regress Epithelial Cancer.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26176230","citation_count":34,"is_preprint":false},{"pmid":"26799921","id":"PMC_26799921","title":"The plastic cellular states of liver cells: Are EpCAM and Lgr5 fit for purpose?","date":"2016","source":"Hepatology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/26799921","citation_count":33,"is_preprint":false},{"pmid":"29245156","id":"PMC_29245156","title":"Selection and targeting of EpCAM protein by ssDNA aptamer.","date":"2017","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/29245156","citation_count":32,"is_preprint":false},{"pmid":"24201161","id":"PMC_24201161","title":"Clinicopathologic implications of EpCAM and Sox2 expression in breast cancer.","date":"2013","source":"Clinical breast cancer","url":"https://pubmed.ncbi.nlm.nih.gov/24201161","citation_count":32,"is_preprint":false},{"pmid":"30185875","id":"PMC_30185875","title":"EpCAM homo-oligomerization is not the basis for its role in cell-cell adhesion.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/30185875","citation_count":32,"is_preprint":false},{"pmid":"24696843","id":"PMC_24696843","title":"Expression of EpCAM increases in the hepatitis B related and the treatment-resistant hepatocellular carcinoma.","date":"2014","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/24696843","citation_count":32,"is_preprint":false},{"pmid":"30419852","id":"PMC_30419852","title":"Epithelial cell adhesion molecule (EpCAM) is involved in prostate cancer chemotherapy/radiotherapy response in vivo.","date":"2018","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/30419852","citation_count":32,"is_preprint":false},{"pmid":"23378578","id":"PMC_23378578","title":"EpCAM modulates NF-κB signaling and interleukin-8 expression in breast cancer.","date":"2013","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/23378578","citation_count":31,"is_preprint":false},{"pmid":"29192390","id":"PMC_29192390","title":"EpCAM-based assays for epithelial tumor cell detection in cerebrospinal fluid.","date":"2017","source":"Journal of neuro-oncology","url":"https://pubmed.ncbi.nlm.nih.gov/29192390","citation_count":30,"is_preprint":false},{"pmid":"31876148","id":"PMC_31876148","title":"Quantitatively Mapping the Assembly Pattern of EpCAM on Cell Membranes with Peptide Probes.","date":"2020","source":"Analytical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31876148","citation_count":30,"is_preprint":false},{"pmid":"38791091","id":"PMC_38791091","title":"Regulation of the Function and Expression of EpCAM.","date":"2024","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/38791091","citation_count":29,"is_preprint":false},{"pmid":"32326212","id":"PMC_32326212","title":"Matriptase Cleaves EpCAM and TROP2 in Keratinocytes, Destabilizing Both Proteins and Associated Claudins.","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32326212","citation_count":29,"is_preprint":false},{"pmid":"23830302","id":"PMC_23830302","title":"High EpCAM expression is linked to proliferation and lauren classification in gastric cancer.","date":"2013","source":"BMC research notes","url":"https://pubmed.ncbi.nlm.nih.gov/23830302","citation_count":29,"is_preprint":false},{"pmid":"29759567","id":"PMC_29759567","title":"EpCAM duality becomes this molecule in a new Dr. Jekyll and Mr. Hyde tale.","date":"2018","source":"Critical reviews in oncology/hematology","url":"https://pubmed.ncbi.nlm.nih.gov/29759567","citation_count":28,"is_preprint":false},{"pmid":"25966221","id":"PMC_25966221","title":"Expression of EpCAM and Wnt/ β-catenin in human colon cancer.","date":"2015","source":"Genetics and molecular research : GMR","url":"https://pubmed.ncbi.nlm.nih.gov/25966221","citation_count":28,"is_preprint":false},{"pmid":"30898885","id":"PMC_30898885","title":"MM-131, a bispecific anti-Met/EpCAM mAb, inhibits HGF-dependent and HGF-independent Met signaling through concurrent binding to EpCAM.","date":"2019","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/30898885","citation_count":28,"is_preprint":false},{"pmid":"28360395","id":"PMC_28360395","title":"Cooverexpression of EpCAM and c-myc genes in malignant breast tumours.","date":"2017","source":"Journal of genetics","url":"https://pubmed.ncbi.nlm.nih.gov/28360395","citation_count":27,"is_preprint":false},{"pmid":"31035965","id":"PMC_31035965","title":"CD44, TGM2 and EpCAM as novel plasma markers in endometrial cancer diagnosis.","date":"2019","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/31035965","citation_count":26,"is_preprint":false},{"pmid":"26543332","id":"PMC_26543332","title":"Circulating tumor cells isolation: the \"post-EpCAM era\".","date":"2015","source":"Chinese journal of cancer research = Chung-kuo yen cheng yen chiu","url":"https://pubmed.ncbi.nlm.nih.gov/26543332","citation_count":25,"is_preprint":false},{"pmid":"31806375","id":"PMC_31806375","title":"EpCAM associates with integrin and regulates cell adhesion in cancer cells.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31806375","citation_count":24,"is_preprint":false},{"pmid":"28315854","id":"PMC_28315854","title":"Mutation of N-linked glycosylation in EpCAM affected cell adhesion in breast cancer cells.","date":"2017","source":"Biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/28315854","citation_count":24,"is_preprint":false},{"pmid":"28560675","id":"PMC_28560675","title":"Enrichment, Isolation and Molecular Characterization of EpCAM-Negative Circulating Tumor Cells.","date":"2017","source":"Advances in experimental medicine and biology","url":"https://pubmed.ncbi.nlm.nih.gov/28560675","citation_count":24,"is_preprint":false},{"pmid":"33490064","id":"PMC_33490064","title":"EpCAM-Mediated Cellular Plasticity Promotes Radiation Resistance and Metastasis in Breast Cancer.","date":"2021","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/33490064","citation_count":24,"is_preprint":false},{"pmid":"27038681","id":"PMC_27038681","title":"Expression of CEA, CA19-9, CA125, and EpCAM in pseudomyxoma peritonei.","date":"2016","source":"Human pathology","url":"https://pubmed.ncbi.nlm.nih.gov/27038681","citation_count":23,"is_preprint":false},{"pmid":"22328825","id":"PMC_22328825","title":"EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy.","date":"2012","source":"Molecular vision","url":"https://pubmed.ncbi.nlm.nih.gov/22328825","citation_count":23,"is_preprint":false},{"pmid":"31354723","id":"PMC_31354723","title":"Immunocyte Profiling Using Single-Cell Mass Cytometry Reveals EpCAM+ CD4+ T Cells Abnormal in Colon Cancer.","date":"2019","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/31354723","citation_count":23,"is_preprint":false},{"pmid":"25550831","id":"PMC_25550831","title":"Overexpression of EpCAM and Trop2 in pituitary adenomas.","date":"2014","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/25550831","citation_count":22,"is_preprint":false},{"pmid":"32392820","id":"PMC_32392820","title":"Feasibility of Imaging EpCAM Expression in Ovarian Cancer Using Radiolabeled DARPin Ec1.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32392820","citation_count":22,"is_preprint":false},{"pmid":"32486423","id":"PMC_32486423","title":"Current View on EpCAM Structural Biology.","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32486423","citation_count":21,"is_preprint":false},{"pmid":"18508569","id":"PMC_18508569","title":"EpCAM in morphogenesis.","date":"2008","source":"Frontiers in bioscience : a journal and virtual library","url":"https://pubmed.ncbi.nlm.nih.gov/18508569","citation_count":21,"is_preprint":false},{"pmid":"29435146","id":"PMC_29435146","title":"Upregulation of LncRNA BCYRN1 promotes tumor progression and enhances EpCAM expression in gastric carcinoma.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29435146","citation_count":21,"is_preprint":false},{"pmid":"30862714","id":"PMC_30862714","title":"Genome-Wide RNAi Screen Identifies PMPCB as a Therapeutic Vulnerability in EpCAM+ Hepatocellular Carcinoma.","date":"2019","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/30862714","citation_count":21,"is_preprint":false},{"pmid":"24566863","id":"PMC_24566863","title":"EpCAM, a potential therapeutic target for esophageal squamous cell carcinoma.","date":"2014","source":"Annals of surgical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/24566863","citation_count":20,"is_preprint":false},{"pmid":"24906438","id":"PMC_24906438","title":"The overexpression of epithelial cell adhesion molecule (EpCAM) in glioma.","date":"2014","source":"Journal of neuro-oncology","url":"https://pubmed.ncbi.nlm.nih.gov/24906438","citation_count":20,"is_preprint":false},{"pmid":"35735360","id":"PMC_35735360","title":"Development of a Novel Anti-EpCAM Monoclonal Antibody for Various Applications.","date":"2022","source":"Antibodies (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/35735360","citation_count":19,"is_preprint":false},{"pmid":"21710497","id":"PMC_21710497","title":"EpCAM- and EGFR-targeted selective gene therapy for biliary cancers using Z33-fiber-modified adenovirus.","date":"2011","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/21710497","citation_count":19,"is_preprint":false},{"pmid":"32961790","id":"PMC_32961790","title":"EpCAM as Modulator of Tissue Plasticity.","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32961790","citation_count":19,"is_preprint":false},{"pmid":"10728619","id":"PMC_10728619","title":"Evaluating GA733-2 mRNA as a marker for the detection of micrometastatic breast cancer in peripheral blood and bone marrow.","date":"1999","source":"Archives of gynecology and obstetrics","url":"https://pubmed.ncbi.nlm.nih.gov/10728619","citation_count":18,"is_preprint":false},{"pmid":"37543570","id":"PMC_37543570","title":"Epithelial cell adhesion molecule (EpCAM) regulates HGFR signaling to promote colon cancer progression and metastasis.","date":"2023","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/37543570","citation_count":18,"is_preprint":false},{"pmid":"12844224","id":"PMC_12844224","title":"Expression of MUC 1 and Ep-CAM in Merkel cell carcinomas: implications for immunotherapy.","date":"2003","source":"Archives of dermatological research","url":"https://pubmed.ncbi.nlm.nih.gov/12844224","citation_count":18,"is_preprint":false},{"pmid":"30116994","id":"PMC_30116994","title":"Drug-induced expression of EpCAM contributes to therapy resistance in esophageal adenocarcinoma.","date":"2018","source":"Cellular oncology (Dordrecht, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/30116994","citation_count":17,"is_preprint":false},{"pmid":"32284393","id":"PMC_32284393","title":"The Biodistribution of a CD3 and EpCAM Bispecific T-Cell Engager Is Driven by the CD3 Arm.","date":"2020","source":"Journal of nuclear medicine : official publication, Society of Nuclear Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/32284393","citation_count":16,"is_preprint":false},{"pmid":"25502397","id":"PMC_25502397","title":"EpCAM knockdown alters microRNA expression in retinoblastoma--functional implication of EpCAM regulated miRNA in tumor progression.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25502397","citation_count":14,"is_preprint":false},{"pmid":"35454789","id":"PMC_35454789","title":"Combined Targeting of AKT and mTOR Inhibits Tumor Formation of EpCAM+ and CD90+ Human Hepatocellular Carcinoma Cells in an Orthotopic Mouse Model.","date":"2022","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/35454789","citation_count":14,"is_preprint":false},{"pmid":"28553629","id":"PMC_28553629","title":"EPCAM Expression in Colon Adenocarcinoma and its Relationship with TNM Staging.","date":"2017","source":"Advanced biomedical research","url":"https://pubmed.ncbi.nlm.nih.gov/28553629","citation_count":13,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":49589,"output_tokens":5282,"usd":0.113999},"stage2":{"model":"claude-opus-4-6","input_tokens":8895,"output_tokens":3850,"usd":0.211088},"total_usd":0.325087,"stage1_batch_id":"msgbatch_0118RScg3WGaGkaZRtWZbtce","stage2_batch_id":"msgbatch_01Cpfzvtbj42aqp5oYZ5Te2s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2009,\n      \"finding\": \"EpCAM undergoes regulated intramembrane proteolysis (RIP): TACE sequentially sheds the ectodomain (EpEX) and presenilin-2 cleaves the remaining stub, releasing the intracellular domain (EpICD). EpICD then associates with FHL2, β-catenin, and Lef-1 to form a nuclear complex that contacts DNA at Lef-1 consensus sites, induces target gene transcription (including c-myc), and is oncogenic in immunodeficient mice.\",\n      \"method\": \"Pharmacological inhibition and genetic silencing of TACE and presenilin-2; nuclear fractionation; Co-IP of EpICD/FHL2/β-catenin/Lef-1 complex; reporter assays; xenograft tumor formation\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods (inhibitor + siRNA + co-IP + reporter + in vivo), strong mechanistic resolution, high citation count\",\n      \"pmids\": [\"19136966\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"The intracellular domain of EpCAM is necessary and sufficient to upregulate c-myc and cyclins A/E, reduce growth-factor dependence, and enhance colony formation, establishing a direct link between EpCAM intracellular signaling and cell-cycle progression.\",\n      \"method\": \"Domain-swapping experiments; antisense-mediated knockdown; metabolic activity and colony formation assays in human 293 cells and NIH3T3 fibroblasts\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — domain-swap mutagenesis with loss-of-function and gain-of-function, replicated in two cell systems\",\n      \"pmids\": [\"15195135\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Silencing EpCAM with siRNA decreases breast cancer cell proliferation, migration, and invasion, and increases the detergent-insoluble (membrane-associated) fractions of E-cadherin, α-catenin, and β-catenin, indicating EpCAM negatively regulates E-cadherin-mediated adhesion complexes.\",\n      \"method\": \"siRNA knockdown; proliferation, migration, and invasion assays; detergent-solubility fractionation of cadherin/catenin complexes\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KD with multiple phenotypic readouts in four cell lines, single lab\",\n      \"pmids\": [\"15313925\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Initial activation of EpCAM cleavage requires cell-to-cell contact (juxtacrine signaling); once EpEX is released it can act in a paracrine manner. EpICD-driven nuclear translocation and c-myc induction require the cleavage step and subsequent nuclear import.\",\n      \"method\": \"Confocal microscopy; immunoblotting; conditional cell-density experiments; conditional EpICD expression system\",\n      \"journal\": \"BMC cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic dissection with cell-contact manipulation and conditional expression, single lab\",\n      \"pmids\": [\"19925656\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"EpCAM promotes cell-cycle progression by transcriptionally upregulating cyclin D1 in a manner dependent on its direct interaction partner FHL2; downstream consequences include Rb phosphorylation and induction of cyclins E and A.\",\n      \"method\": \"EpCAM overexpression and knockdown; cyclin D1 promoter assays; co-IP of EpICD–FHL2; immunohistochemistry for Ki67, cyclin D1, and phospho-Rb in patient tumors\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reporter assay + co-IP + in vivo correlation, moderate evidence from single lab with orthogonal methods\",\n      \"pmids\": [\"22391566\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EpCAM physically associates with claudin-7 (and claudin-1 via claudin-7) and protects them from lysosomal degradation; EpCAM knockdown reduces claudin-7 and claudin-1 protein levels and paradoxically increases their accumulation at tight junctions, altering trans-epithelial resistance.\",\n      \"method\": \"Preparative immunoprecipitation; co-IP; shRNA knockdown; trans-epithelial electroresistance measurements; immunofluorescence microscopy with lysosome inhibitors\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP + functional rescue with lysosome inhibitors + shRNA phenotype, strong evidence from multiple orthogonal approaches\",\n      \"pmids\": [\"23486470\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"EpCAM–claudin-7 interaction requires an AxxxG motif in EpCAM's transmembrane region; the resulting complex is recruited into tetraspanin-enriched membrane microdomains (TEMs) where it supports sustained ERK1/2 phosphorylation, upregulation of anti-apoptotic proteins, drug resistance, and enhanced cell motility, but abolishes EpCAM-mediated homophilic adhesion.\",\n      \"method\": \"Transfection of deletion and point-mutant EpCAM/claudin-7 constructs in HEK293 and BSp73AS cells; co-IP; TEM fractionation; proliferation, migration, and tumorigenicity assays\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — mutagenesis identifying specific transmembrane motif + co-IP + TEM fractionation + functional assays\",\n      \"pmids\": [\"19276185\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EpCAM acts as a potent inhibitor of novel PKC (nPKC) via a short pseudosubstrate-like segment in its cytoplasmic tail that binds nPKCs with high affinity; loss of EpCAM in Xenopus embryos leads to PKC overstimulation, ERK pathway hyperactivation, exacerbated myosin contractility, loss of cadherin-mediated adhesion, and tissue dissociation.\",\n      \"method\": \"Xenopus embryo loss-of-function; in vitro PKC binding assay with cytoplasmic tail peptides; mutagenesis; myosin contractility measurements; cadherin adhesion assays\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro binding assay with mutagenesis + in vivo phenotype in Xenopus, strong mechanistic resolution\",\n      \"pmids\": [\"24183651\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Matriptase cleaves EpCAM after Arg80; loss of the matriptase inhibitor HAI-2 in intestinal epithelial cells leads to unrestrained matriptase activity, efficient EpCAM cleavage, reduced EpCAM–claudin-7 association, and lysosomal degradation of both EpCAM and claudin-7, causing intestinal epithelial dysplasia (congenital tufting enteropathy phenotype).\",\n      \"method\": \"In vitro cleavage of purified recombinant proteins; cell transfection; HAI-2 knockout mouse model; immunoprecipitation; HAI-2 CTE mutant functional rescue experiments\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution + mutagenesis + knockout mouse model with defined disease phenotype, strong mechanistic resolution\",\n      \"pmids\": [\"28094766\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EpCAM homo-oligomerization does not mediate intercellular adhesion; EpCAM monomers do not form inter-cellular oligomers detectable by SAXS or XL-MS, and bead aggregation assays confirm no homophilic adhesion. EpCAM forms stable cis-dimers on the cell surface at pre-formed cell–cell contacts as detected by FLIM-FRET.\",\n      \"method\": \"SAXS; cross-linking mass spectrometry (XL-MS); bead aggregation assay; FLIM-FRET on live cells\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — multiple biophysical methods (SAXS, XL-MS, FLIM-FRET, bead aggregation) in one study\",\n      \"pmids\": [\"30185875\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"The extracellular domain of EpCAM (EpEX) activates EGFR and downstream ERK1/2 signaling to promote colon cancer cell migration and proliferation; EpEX–EGFR–ERK1/2 signaling positively regulates RIP of EpCAM and EpICD shedding, which then drives nuclear β-catenin accumulation and HIF1α target gene expression.\",\n      \"method\": \"EGFR/MEK inhibitor treatment; EpEX-stimulation assays; immunoprecipitation; Western blotting; xenograft tumor models; analysis of colorectal cancer tissues\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological inhibition + IP + in vivo validation, single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"29981429\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"EpCAM and its complex partner claudin-7 (Cldn7) facilitate transcription-factor-mediated somatic cell reprogramming to iPSCs; overexpression of EpCAM or EpICD enhances reprogramming efficiency, activates the Oct4 promoter, and suppresses p53 and p21 expression.\",\n      \"method\": \"Quantitative RT-PCR; alkaline phosphatase and Nanog colony assays; EpCAM/EpICD overexpression; shRNA knockdown; Oct4 promoter reporter assay; iPSC characterization in vitro and in vivo\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — gain/loss-of-function with promoter reporter and iPSC functional characterization, single lab\",\n      \"pmids\": [\"21799003\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Mutations in EPCAM (including a splice-site mutation causing deletion of exon 4) are the genetic cause of congenital tufting enteropathy, resulting in loss of EpCAM protein from intestinal epithelium.\",\n      \"method\": \"SNP homozygosity mapping; direct DNA sequencing; RT-PCR; immunohistochemistry; Western blotting of patient intestinal tissue\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic cause established by sequencing + protein confirmation in patient tissue, replicated in multiple unrelated patients\",\n      \"pmids\": [\"18572020\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"3′-end deletions of EPCAM encompassing the transcription termination signal cause allele-specific epigenetic silencing (promoter hypermethylation) of the neighboring DNA mismatch repair gene MSH2 in tissues expressing EPCAM, leading to Lynch syndrome.\",\n      \"method\": \"Multiplex ligation-dependent probe amplification (MLPA); MSH2 promoter methylation analysis; tissue-specific expression profiling in germline deletion carriers\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — mechanism (read-through transcription silencing MSH2) confirmed in 45 families with methylation analysis, strong replicated evidence\",\n      \"pmids\": [\"21309036\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EpCAM mutation (exon 4 deletion) causes loss of EpCAM–claudin-7 colocalization and complex formation in vivo, leading to intestinal barrier dysfunction (increased permeability, reduced ion transport, altered desmosomes) and the CTE pathological phenotype.\",\n      \"method\": \"Cre-LoxP conditional mouse model; histology; electron microscopy; immunohistochemistry; barrier permeability assays; ion transport measurements; comparison with CTE patient tissue\",\n      \"journal\": \"American journal of physiology. Gastrointestinal and liver physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo knockout mouse model with defined molecular (EpCAM/claudin-7 complex disruption) and functional (barrier/transport) phenotype, validated in human CTE tissue\",\n      \"pmids\": [\"24337010\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"EpCAM signaling activates NF-κB, promotes c-Jun phosphorylation via the JNK pathway, and increases AP-1 transcription factor activity; soluble extracellular EpCAM (rEpEX) can rescue invasion, AP-1 activity, and c-Jun phosphorylation after EpCAM ablation.\",\n      \"method\": \"RNAi loss-of-function; gain-of-function with cDNA construct; recombinant EpEX rescue; phosphoprotein analysis; AP-1 and JNK reporter/inhibitor experiments; invasion assays in vitro and in vivo\",\n      \"journal\": \"Breast cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — RNAi + constitutively active constructs + pharmacological inhibitors + rescue experiments, single lab\",\n      \"pmids\": [\"22132731\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EpCAM ablation decreases NF-κB transcription factor activity and RELA phosphorylation, increases IκBα protein, and reduces IL-8 expression; forced expression of IκBα or RELA ablation blocks EpCAM-dependent rescue of IL-8 promoter activity.\",\n      \"method\": \"RNAi; constitutively active/dominant negative NF-κB constructs; IL-8 promoter reporter assay; ELISA; Western blotting\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reporter assay + genetic rescue experiments, single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"23378578\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"EpCAM associates with integrin β1 (shown by co-immunoprecipitation); EpCAM knockout reduces integrin α5 expression and decreases phosphorylation of FAK, AKT, and ERK, impairing cell adhesion, migration, and proliferation on extracellular matrix proteins.\",\n      \"method\": \"CRISPR/Cas9 EpCAM knockout; co-immunoprecipitation; Western blotting for FAK/AKT/ERK phosphorylation; adhesion and migration assays\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — CRISPR KO with co-IP and signaling readouts, single lab\",\n      \"pmids\": [\"31806375\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EpCAM organizes into heterogeneous clusters on the cell membrane and localizes within tetraspanin-enriched microdomains (TEMs) through co-association with CD9; cytoskeleton integrity and N-glycosylation both limit EpCAM cluster assembly.\",\n      \"method\": \"Direct stochastic optical reconstruction microscopy (dSTORM) with fluorophore-conjugated peptides; dual-color super-resolution imaging; CD9 knockdown; cytoskeleton disruption; glycosylation inhibition\",\n      \"journal\": \"Analytical chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 — super-resolution imaging with single-molecule precision + genetic/pharmacological perturbations, single lab\",\n      \"pmids\": [\"31876148\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Matriptase cleaves both EpCAM and its homolog TROP2 in keratinocytes; EpCAM and TROP2 are redundant in stabilizing claudin-1 and claudin-7 (knockdown of either alone has small effects, but combined knockdown markedly reduces claudins); HAI-1 (not HAI-2) is the principal physiological inhibitor of matriptase in keratinocytes.\",\n      \"method\": \"In vitro cleavage of purified recombinant proteins; siRNA knockdown of EpCAM, TROP2, HAI-1, HAI-2, matriptase in HaCaT cells; Western blotting; lysosome inhibitor (chloroquine) rescue\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with purified proteins + cell-based genetic validation with multiple orthogonal siRNA knockdowns\",\n      \"pmids\": [\"32326212\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The extracellular domain of EpCAM (EpEX) binds directly to HGFR (c-Met) and cooperates with HGF to activate downstream ERK and FAK-AKT signaling; EpEX stabilizes β-catenin and Snail by decreasing GSK3β activity, promoting EMT and metastasis in colon cancer.\",\n      \"method\": \"Immunoprecipitation; ELISA; FRET; Western blotting; migration/invasion assays; tail vein injection and orthotopic metastasis mouse models\",\n      \"journal\": \"Journal of translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP/FRET demonstrating direct EpEX–HGFR interaction + in vivo metastasis model, single lab\",\n      \"pmids\": [\"37543570\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EpCAM is a multifunctional transmembrane glycoprotein that undergoes regulated intramembrane proteolysis (by TACE and presenilin-2, initiated by cell-to-cell contact) to release a soluble ectodomain (EpEX) and a nuclear intracellular domain (EpICD); EpICD forms a complex with FHL2, β-catenin, and Lef-1 to drive transcription of proliferative target genes (c-myc, cyclin D1); EpEX acts as a ligand for EGFR and HGFR to activate ERK, AKT, and downstream pathways; EpCAM also directly inhibits novel PKC via a pseudosubstrate motif in its cytoplasmic tail to suppress actomyosin contractility, interacts with claudin-7 and claudin-1 to stabilize them from lysosomal degradation (a function regulated by the matriptase/HAI protease axis), and associates with integrin β1 to modulate FAK/ERK signaling, while germline mutations cause congenital tufting enteropathy and 3′ deletions silence MSH2 to cause Lynch syndrome.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"EpCAM is a transmembrane glycoprotein that functions as a signaling hub in epithelial proliferation, adhesion, and differentiation through regulated intramembrane proteolysis (RIP) and direct modulation of multiple signaling pathways. TACE-mediated ectodomain shedding followed by presenilin-2 cleavage releases a soluble ectodomain (EpEX) that activates EGFR and HGFR to stimulate ERK, AKT, and FAK signaling, and an intracellular domain (EpICD) that forms a nuclear complex with FHL2, β-catenin, and Lef-1 to drive transcription of c-myc and cyclin D1, promoting cell-cycle progression [PMID:19136966, PMID:15195135, PMID:22391566, PMID:29981429, PMID:37543570]. EpCAM directly inhibits novel PKC isoforms via a pseudosubstrate motif in its cytoplasmic tail, thereby restraining actomyosin contractility and maintaining cadherin-mediated adhesion, and it physically associates with claudin-7 through a transmembrane AxxxG motif to protect claudins from matriptase-initiated lysosomal degradation — a mechanism whose disruption causes congenital tufting enteropathy [PMID:24183651, PMID:23486470, PMID:19276185, PMID:28094766, PMID:18572020]. Germline 3′ deletions of EPCAM additionally cause epigenetic silencing of the adjacent MSH2 gene, resulting in Lynch syndrome [PMID:21309036].\",\n  \"teleology\": [\n    {\n      \"year\": 2004,\n      \"claim\": \"Establishing that EpCAM is not merely an adhesion molecule but an active signaling receptor whose intracellular domain is necessary and sufficient for upregulating c-myc and cyclins and promoting proliferation answered the fundamental question of whether EpCAM has cell-autonomous oncogenic signaling capacity.\",\n      \"evidence\": \"Domain-swap mutagenesis with gain/loss-of-function in HEK293 and NIH3T3 cells; siRNA knockdown in breast cancer lines with proliferation, migration, and cadherin fractionation readouts\",\n      \"pmids\": [\"15195135\", \"15313925\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Protease responsible for EpICD release unknown\", \"Identity of nuclear partners of EpICD not yet defined\", \"Mechanism linking EpCAM to E-cadherin regulation unclear\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identification of homozygous EPCAM mutations as the genetic cause of congenital tufting enteropathy established EpCAM as essential for intestinal epithelial integrity and linked it to a Mendelian disease.\",\n      \"evidence\": \"SNP homozygosity mapping and sequencing in multiple unrelated CTE families; immunohistochemistry confirming protein loss in patient intestinal tissue\",\n      \"pmids\": [\"18572020\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism of epithelial disruption upon EpCAM loss not defined\", \"Claudin interactions not yet identified\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Discovery of the complete regulated intramembrane proteolysis (RIP) cascade — TACE ectodomain shedding followed by presenilin-2 cleavage releasing EpICD, which assembles a nuclear FHL2/β-catenin/Lef-1 transcription complex — provided the full mechanistic pathway from membrane to gene activation.\",\n      \"evidence\": \"Pharmacological and siRNA inhibition of TACE and presenilin-2; nuclear fractionation and co-IP of EpICD complex; reporter assays; xenograft tumorigenesis; cell-contact density experiments showing juxtacrine initiation\",\n      \"pmids\": [\"19136966\", \"19925656\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Crystal structure of EpICD-containing nuclear complex unavailable\", \"Upstream signals triggering TACE activation at cell contacts not defined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Mapping the EpCAM–claudin-7 interaction to a transmembrane AxxxG motif and demonstrating that this complex is recruited to tetraspanin-enriched microdomains (TEMs) where it sustains ERK1/2 signaling and drug resistance revealed a second, adhesion-independent oncogenic function of EpCAM.\",\n      \"evidence\": \"Deletion and point-mutant constructs in HEK293/BSp73AS cells; co-IP; TEM fractionation; proliferation, migration, and tumorigenicity assays\",\n      \"pmids\": [\"19276185\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the AxxxG-mediated transmembrane interaction not resolved\", \"Relative contributions of RIP signaling versus TEM-based signaling to tumorigenesis unclear\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Demonstration that 3′ EPCAM deletions cause read-through transcription and epigenetic silencing of MSH2 defined a cis-regulatory mechanism for Lynch syndrome independent of MSH2 coding mutations.\",\n      \"evidence\": \"MLPA and MSH2 promoter methylation analysis in 45 carrier families; tissue-specific expression profiling\",\n      \"pmids\": [\"21309036\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Precise chromatin mechanism of read-through-induced silencing not fully dissected\", \"Genotype–phenotype correlation for different deletion sizes incomplete\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identification of cyclin D1 as a direct transcriptional target of EpICD–FHL2 signaling, with consequent Rb phosphorylation, connected EpCAM proteolytic signaling to a canonical cell-cycle progression mechanism.\",\n      \"evidence\": \"EpCAM overexpression/knockdown; cyclin D1 promoter reporter; co-IP of EpICD–FHL2; Ki67 and cyclin D1 IHC in patient tumors\",\n      \"pmids\": [\"22391566\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full set of EpICD transcriptional targets undefined\", \"Chromatin occupancy by the EpICD complex not mapped genome-wide\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Discovery that EpCAM's cytoplasmic tail contains a pseudosubstrate motif that directly inhibits novel PKC isoforms, thereby restraining myosin contractility and preserving cadherin adhesion, established a proteolysis-independent signaling function for full-length EpCAM.\",\n      \"evidence\": \"In vitro PKC binding assay with synthetic cytoplasmic-tail peptides; mutagenesis; Xenopus embryo loss-of-function with actomyosin and cadherin phenotyping\",\n      \"pmids\": [\"24183651\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether nPKC inhibition is operative in mammalian epithelia not confirmed\", \"Relative importance of PKC-inhibitory versus RIP functions in CTE pathogenesis unknown\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Showing that EpCAM protects claudin-7 and claudin-1 from lysosomal degradation, and that CTE-causing EpCAM mutations ablate EpCAM–claudin-7 complex formation causing intestinal barrier dysfunction, provided a molecular explanation for congenital tufting enteropathy.\",\n      \"evidence\": \"Reciprocal co-IP; shRNA knockdown with lysosome inhibitor rescue; conditional EpCAM-knockout mouse; electron microscopy; barrier permeability and ion transport assays; comparison with CTE patient tissue\",\n      \"pmids\": [\"23486470\", \"24337010\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether claudin destabilization alone accounts for the full CTE phenotype is untested\", \"Role of desmosome alterations observed in KO mice not mechanistically dissected\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identification of matriptase as the protease cleaving EpCAM at Arg80, with HAI-2 as its inhibitor, showed that uncontrolled matriptase activity phenocopies CTE by destabilizing the EpCAM–claudin-7 complex via ectodomain cleavage.\",\n      \"evidence\": \"In vitro cleavage with purified proteins; HAI-2 knockout mouse; co-IP; CTE mutant rescue\",\n      \"pmids\": [\"28094766\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether matriptase cleavage feeds into RIP-dependent EpICD signaling or only triggers degradation is unclear\", \"Tissue-specific regulation of matriptase/HAI balance not mapped comprehensively\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Biophysical evidence that EpCAM does not mediate intercellular homophilic adhesion but forms cis-dimers at pre-existing contacts overturned the long-held model of EpCAM as a cell-adhesion molecule and reframed it primarily as a signaling receptor.\",\n      \"evidence\": \"SAXS, cross-linking mass spectrometry, bead aggregation assay, and FLIM-FRET on live cells\",\n      \"pmids\": [\"30185875\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional role of cis-dimerization not established\", \"Whether cis-dimers are the signaling-competent unit for RIP is untested\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Showing that the shed ectodomain EpEX activates EGFR–ERK1/2 signaling to promote proliferation and migration, and that this pathway feeds back to stimulate further EpCAM RIP and nuclear β-catenin accumulation, established a positive-feedback loop linking extracellular and intracellular EpCAM signals.\",\n      \"evidence\": \"EGFR/MEK inhibitor treatment; EpEX stimulation; co-IP; xenograft models; colorectal cancer tissue analysis\",\n      \"pmids\": [\"29981429\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct EpEX–EGFR binding site not mapped\", \"Relative importance of autocrine versus paracrine EpEX signaling in vivo unknown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Super-resolution imaging revealed that EpCAM forms heterogeneous nanoclusters within CD9-positive tetraspanin microdomains, regulated by cytoskeletal integrity and N-glycosylation, providing a spatial framework for its signaling functions. Parallel work confirmed functional redundancy between EpCAM and TROP2 in stabilizing claudins against matriptase-mediated degradation.\",\n      \"evidence\": \"dSTORM with CD9 knockdown and pharmacological perturbations; in vitro cleavage assays with siRNA knockdown of EpCAM/TROP2/matriptase/HAI-1/HAI-2 in keratinocytes\",\n      \"pmids\": [\"31876148\", \"32326212\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How nanocluster organization relates to RIP activation is unknown\", \"TROP2 redundancy complicates tissue-specific phenotype predictions\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstration that EpEX directly binds HGFR (c-Met) and cooperates with HGF to activate ERK and FAK-AKT signaling, stabilize β-catenin/Snail, and promote EMT and metastasis expanded the receptor repertoire of shed EpEX beyond EGFR.\",\n      \"evidence\": \"Co-IP, FRET, ELISA for direct EpEX–HGFR interaction; migration/invasion assays; orthotopic and tail-vein metastasis mouse models\",\n      \"pmids\": [\"37543570\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"EpEX binding site on HGFR not mapped structurally\", \"Whether EpEX activates additional RTKs is unknown\", \"Relative contribution of EGFR versus HGFR arm in vivo not delineated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include how cis-dimerization and nanocluster organization couple to RIP initiation, the genome-wide transcriptional program of the EpICD nuclear complex, and how the PKC-inhibitory and claudin-stabilizing functions of full-length EpCAM are coordinated with proteolytic signaling in different epithelial contexts.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No ChIP-seq or CUT&RUN mapping of EpICD-containing complex\", \"Structural basis of EpICD–FHL2–β-catenin–Lef-1 assembly unknown\", \"Relative contribution of PKC inhibition versus RIP signaling to epithelial homeostasis not dissected in mammalian models\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 1, 4]},\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [10, 20]},\n      {\"term_id\": \"GO:0098631\", \"supporting_discovery_ids\": [5, 14]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6, 9, 18]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 3, 4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 7, 10, 15, 17, 20]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [1, 4]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0, 4, 16]},\n      {\"term_id\": \"R-HSA-1500931\", \"supporting_discovery_ids\": [5, 14]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [12, 13]}\n    ],\n    \"complexes\": [\n      \"EpICD/FHL2/β-catenin/Lef-1 nuclear transcription complex\",\n      \"EpCAM–claudin-7 membrane complex\",\n      \"Tetraspanin-enriched microdomain (TEM) complex\"\n    ],\n    \"partners\": [\n      \"FHL2\",\n      \"CTNNB1\",\n      \"CLDN7\",\n      \"CLDN1\",\n      \"EGFR\",\n      \"MET\",\n      \"ITGB1\",\n      \"CD9\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}