{"gene":"EPCAM","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":2009,"finding":"EpCAM undergoes regulated intramembrane proteolysis (RIP): its ectodomain EpEX is shed by TACE, and the remaining stub is cleaved by presenilin-2 to release the intracellular domain EpICD, which translocates to the nucleus. Nuclear EpICD forms a complex with FHL2, β-catenin, and Lef-1 that binds Lef-1 consensus DNA sites, drives target gene transcription, and is oncogenic in immunodeficient mice. Pharmacological or genetic inhibition of either protease impairs EpCAM-dependent proliferative signaling.","method":"Pharmacological inhibition (TACE/presenilin-2 inhibitors), genetic silencing (siRNA), conditional cell systems, confocal microscopy, immunoblotting, nuclear fractionation, xenograft oncogenesis assay, patient tissue immunohistochemistry","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (biochemical cleavage assays, genetic silencing of both proteases, nuclear fractionation, in vivo oncogenesis, patient tissue validation) in a single rigorous study; widely replicated by subsequent papers","pmids":["19136966"],"is_preprint":false},{"year":2009,"finding":"Activation of EpCAM's RIP-dependent oncogenic signaling requires cell-to-cell contact: when intercellular contact is prevented, EpCAM does not confer growth advantage. Contact triggers initial cleavage (juxtacrine), releasing soluble EpEX that can act in a paracrine manner. The pre-cleaved EpICD fragment bypasses the contact requirement but still requires nuclear translocation to induce c-myc and proliferation.","method":"Density-dependent cell culture experiments, conditional cell systems expressing pre-cleaved EpICD, confocal laser scanning microscopy, immunoblotting, cell counting, c-myc reporter assays","journal":"BMC cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple complementary cell-based assays in a single lab; consistent with the RIP mechanism established in PMID 19136966","pmids":["19925656"],"is_preprint":false},{"year":2012,"finding":"EpCAM drives cell cycle progression by upregulating cyclin D1 transcription in an FHL2-dependent manner. Downstream consequences include phosphorylation of retinoblastoma protein (Rb) and induction of cyclins E and A. In vivo, EpCAM expression level positively correlates with Ki67, nuclear cyclin D1, and Rb phosphorylation.","method":"EpCAM knockdown/overexpression in cancer cells, RT-PCR and Western blotting for cyclins, Rb phosphorylation assays, FHL2 co-depletion rescue experiments, immunohistochemistry of patient tissues","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic manipulation with defined molecular readouts, corroborated by in vivo IHC, single lab","pmids":["22391566"],"is_preprint":false},{"year":2013,"finding":"EpCAM physically associates with claudin-7 (tight interaction) and, indirectly via claudin-7, with claudin-1, but not claudin-2 or claudin-4. This interaction stabilizes claudin-7 and claudin-1 by preventing their lysosomal degradation. EpCAM knockdown reduces claudin-7 and claudin-1 protein levels, which are restored by lysosomal inhibitors; this also paradoxically increases claudin accumulation at tight junctions and enhances transepithelial electroresistance.","method":"shRNA knockdown of EpCAM in T84 and Caco-2 cells, preparative immunoprecipitation, co-immunoprecipitation, co-transfection experiments, lysosomal inhibitor rescue, immunofluorescence microscopy, transepithelial electrical resistance measurement","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — reciprocal co-IP, preparative IP, genetic knockdown, lysosomal rescue, functional TJ assays; multiple orthogonal methods in one rigorous study","pmids":["23486470"],"is_preprint":false},{"year":2013,"finding":"EpCAM acts as a potent inhibitor of novel protein kinase C (nPKC) through a short pseudosubstrate-like motif in its cytoplasmic tail that binds nPKCs with high affinity. Loss of EpCAM in amphibian embryos sequentially overstimulates PKC, activates the Erk pathway, exacerbates myosin contractility, disrupts cadherin-mediated adhesion, and causes tissue dissociation. This PKC-inhibitory mechanism is shared with other plasma membrane adhesion molecules.","method":"Loss-of-function in Xenopus embryos, PKC activity assays, Erk pathway measurements, myosin contractility readouts, in vitro binding assays of EpCAM cytoplasmic tail segment to nPKCs, bioinformatics motif analysis","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro binding assay identifying the minimal inhibitory motif, loss-of-function in intact tissue with sequential phenotypic readouts, multiple orthogonal methods","pmids":["24183651"],"is_preprint":false},{"year":2000,"finding":"GA733-2/EpCAM extracellular domain disulfide-bonding pattern was determined: cysteines 1–6 form a novel pattern (Cys1-Cys4, Cys2-Cys6, Cys3-Cys5) distinct from EGF-like structure, and cysteines 7–12 form a thyroglobulin type 1A pattern (Cys7-Cys8, Cys9-Cys10, Cys11-Cys12). N-linked glycosylation occurs at Asn88 (complete) and Asn51 (partial); Asn175 is not glycosylated; no O-linked carbohydrate was detected.","method":"MALDI mass spectrometry, N-terminal sequencing of tryptic peptides, chemical reduction and alkylation, glycosylation site analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct biochemical structural determination with mass spectrometry and sequencing; constitutes the primary structural characterization of EpCAM's disulfide architecture","pmids":["11080501"],"is_preprint":false},{"year":2000,"finding":"Full-length GA733-2/EpCAM exists as high-affinity non-covalent cis-dimers (Kd <10 nM for monomer-dimer) and lower-affinity tetramers (~10 µM for dimer-tetramer) in solution. The extracellular domain alone is monomeric. Cell-cell adhesion activity resides in the full-length dimer/tetramer; monomeric extracellular domain is inactive in cell aggregation inhibition assays.","method":"Sedimentation equilibrium ultracentrifugation, chemical cross-linking of purified protein and cells in suspension vs. monolayer, cell aggregation inhibition assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — biophysical (sedimentation equilibrium) plus biochemical (cross-linking) plus functional (aggregation) methods establish oligomeric state and link it to adhesion activity","pmids":["11058587"],"is_preprint":false},{"year":2018,"finding":"EpCAM monomers do not associate into intercellular homo-oligomers capable of mediating cell-cell adhesion. EpCAM forms stable cis-dimers on the surface of cells with pre-formed contacts (detected by FLIM-FRET), but no trans-oligomers between opposing cells were detectable. These findings argue against EpCAM functioning as a direct homophilic cell-cell adhesion molecule.","method":"SAXS, cross-linking mass spectrometry (XL-MS), bead aggregation assays, FLIM-FRET on cell surfaces","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1 / Moderate — multiple structural/biophysical methods (SAXS, XL-MS, FLIM-FRET) in one study; result is explicitly a negative finding regarding intercellular homo-oligomerization","pmids":["30185875"],"is_preprint":false},{"year":2013,"finding":"In a mouse model of congenital tufting enteropathy (CTE), deletion of EpCAM exon 4 causes mislocalization of residual mutant EpCAM protein, loss of claudin-7 colocalization, and disruption of the EpCAM/claudin-7 complex, leading to enhanced intestinal permeability and epithelial cell migration. These findings were confirmed in CTE patient intestinal tissue.","method":"Cre-LoxP mouse model (Epcam Δ4/Δ4), histology, light and electron microscopy, immunohistochemistry, permeability assays, co-localization studies in mouse and patient tissue","journal":"American journal of physiology. Gastrointestinal and liver physiology","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo genetic model validated against human patient tissue, with functional permeability/migration readouts and protein complex analysis","pmids":["24337010"],"is_preprint":false},{"year":2014,"finding":"EpCAM mutation (exon 4 deletion) in intestinal epithelial cells leads to decreased electrical resistance, increased paracellular permeability, and decreased ion transport. In the inducible mouse model, EpCAM deficiency also decreases tight junctional protein expression in intestine.","method":"EpCAM shRNA knockdown in T84 colonic epithelial cells, transepithelial electrical resistance measurement, permeability assays, ion transport assays, tamoxifen-inducible EpCAM Δ4/Δ4 mouse model, Western blotting","journal":"Journal of molecular medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — both in vitro (knockdown) and in vivo (inducible mouse model) with quantitative functional barrier and transport readouts","pmids":["25482158"],"is_preprint":false},{"year":2020,"finding":"The EGF-like domain I within the extracellular domain of EpCAM (EpEX) binds EGFR, activating AKT and MAPK signaling. AKT signaling inhibits FOXO3a and stabilizes PD-L1 protein; MAPK signaling also contributes to PD-L1 stabilization. Neutralizing EpCAM (anti-EpCAM antibody EpAb2-6) reduces PD-L1 levels and promotes FOXO3a nuclear translocation and HtrA2-mediated apoptosis.","method":"Co-immunoprecipitation of EpEX with EGFR, Western blotting for AKT/MAPK/FOXO3a/PD-L1, immunofluorescence for FOXO3a localization, in vivo mouse metastasis and orthotopic colorectal cancer models","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP establishing EpEX-EGFR interaction combined with signaling pathway readouts and in vivo validation, single lab","pmids":["32978170"],"is_preprint":false},{"year":2023,"finding":"The extracellular domain of EpCAM (EpEX) binds HGFR (c-Met) and cooperates with HGF to activate downstream ERK and FAK-AKT signaling, stabilizes active β-catenin and Snail by reducing GSK3β activity, and promotes EMT and metastasis of colon cancer cells.","method":"Immunoprecipitation, ELISA, FRET to confirm EpEX-HGFR interaction, Western blotting for downstream signaling, cell proliferation/migration/invasion assays, tail-vein metastasis and orthotopic animal models","journal":"Journal of translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — IP/FRET establishing the EpEX-HGFR interaction plus downstream signaling and in vivo validation, single lab","pmids":["37543570"],"is_preprint":false},{"year":2013,"finding":"EpCAM knockdown in breast cancer cells decreases NF-κB transcription factor activity (reduced RELA phosphorylation, increased IκBα) and reduces IL-8 expression. Rescue experiments show that RELA ablation or forced IκBα expression prevented EpCAM-dependent IL-8 promoter reactivation, placing EpCAM upstream of the NF-κB/RELA/IL-8 axis in breast cancer invasion.","method":"EpCAM siRNA knockdown in breast cancer cell lines, NF-κB reporter assays, RELA phosphorylation Western blotting, IκBα Western blotting, IL-8 ELISA, IL-1β stimulation, functional rescue with RELA siRNA and IκBα overexpression","journal":"Molecular cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockdown with epistatic rescue experiments defining pathway position, single lab","pmids":["23378578"],"is_preprint":false},{"year":2019,"finding":"EpCAM associates with integrin β1 (demonstrated by co-immunoprecipitation). EpCAM knockout (CRISPR/Cas9) in colorectal CW-2 and epidermoid A431 cells reduces integrin α5 expression, decreases FAK, AKT, and ERK phosphorylation, and impairs cell adhesion, migration, and colony formation.","method":"CRISPR/Cas9 knockout, co-immunoprecipitation, Western blotting for phospho-FAK/AKT/ERK, cell adhesion assays, migration assays, colony formation assays","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — co-IP identifying EpCAM–integrin β1 association plus CRISPR KO with defined signaling readouts, single lab","pmids":["31806375"],"is_preprint":false},{"year":2020,"finding":"Matriptase cleaves both EpCAM and TROP2 in keratinocytes (HaCaT cells). Matriptase cleavage of EpCAM and TROP2 destabilizes claudin-1 and claudin-7 by routing them to lysosomes. This cleavage is inhibited by HAI proteins (HAI-1 more than HAI-2); dual knockdown of HAI-1 and HAI-2 causes near-complete cleavage of EpCAM/TROP2 and drastic reduction of claudins, reversible by concurrent matriptase knockdown. Protease-disabled matriptase or G827R mutant does not cleave EpCAM or TROP2.","method":"In vitro cleavage assay with purified recombinant proteins, co-transfection of matriptase in 293T cells, siRNA knockdown of HAI-1/HAI-2/matriptase in HaCaT cells, Western blotting, lysosomal inhibitor (chloroquine) rescue","journal":"Cells","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro reconstitution with purified proteins, active-site mutant controls, cell-based corroboration with epistatic knockdowns, multiple orthogonal methods","pmids":["32326212"],"is_preprint":false},{"year":2022,"finding":"EPCAM and TROP2 share redundant roles in stabilizing claudin-7 at the cell membrane and in epithelial development across multiple tissues. TROP2 can compensate for loss of EPCAM in claudin-7 stabilization in tissues co-expressing both proteins; combined loss causes much more severe developmental defects than either single knockout alone.","method":"Epcam knockout mice, Trop2 knockout mice, Epcam/Trop2 double knockout mice, histology of multiple tissues, immunofluorescence for claudin-7 localization and expression, viability and growth tracking","journal":"Biology open","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis using single and double knockout mice with molecular readout (claudin-7 stabilization), replicated across multiple tissues","pmids":["35730316"],"is_preprint":false},{"year":1994,"finding":"DNA methylation of the TROP1/EpCAM gene locus prevents gene amplification after transfection. Demethylation of TROP1 (by 5-azacytidine treatment) allows efficient amplification of transfected TROP1 copies (up to 40 copies per haploid genome), demonstrating that methylation status controls the amplifiability of this locus.","method":"5-azacytidine demethylation treatment of JAR choriocarcinoma cells, DNA transfection, FACS-based selection, Southern blot copy-number analysis, in vitro Sss I methylase treatment of DNA","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — enzymatic methylation rescue experiment plus demethylation treatment establishing causal link between methylation and amplification resistance, single lab","pmids":["8016075"],"is_preprint":false},{"year":2003,"finding":"Loss of TP53 (p53) induces demethylation of the TROP1/EpCAM gene, leading to TROP1 gene amplification. Reintroduction of wild-type TP53 into p53-null cells reverses the demethylation. In vitro methylation of transfected TROP1 DNA with Sss I methylase prevents amplification, establishing a p53→DNA methylation→TROP1 amplification axis.","method":"TP53 transfection into TP53-null cells, Southern blot for methylation and copy number, in vitro methylation with Sss I methylase, gene amplification assay in vivo","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic rescue (wtTP53 reintroduction) plus in vitro methylation control, single lab","pmids":["12642870"],"is_preprint":false},{"year":2015,"finding":"An anti-EpCAM antibody (EpAb2-6) binding to positions Y95 and D96 of the EGF-II/TY domain of EpCAM inhibits production of EpICD, thereby decreasing its nuclear translocation and downstream signal activation (oncogenic signaling). This antibody induces cancer cell apoptosis in vitro and inhibits tumor growth in vivo.","method":"Antibody epitope mapping, Western blotting for EpICD production and nuclear translocation, cell apoptosis assays, xenograft mouse models","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epitope-defined antibody blocking EpICD generation with downstream signaling readouts, single lab","pmids":["26317650"],"is_preprint":false}],"current_model":"EpCAM is a transmembrane glycoprotein that functions through multiple mechanisms: (1) it undergoes regulated intramembrane proteolysis sequentially by TACE and presenilin-2, releasing soluble EpEX and nuclear EpICD, which complexes with FHL2, β-catenin, and Lef-1 to drive proliferative gene transcription (including cyclin D1 and c-myc) in a cell-contact-initiated manner; (2) its cytoplasmic pseudosubstrate-like motif directly inhibits novel PKC, suppressing actomyosin contractility and promoting tissue plasticity; (3) it stabilizes claudin-1 and claudin-7 by protecting them from lysosomal degradation via physical association—a function shared redundantly with TROP2 and disrupted by matriptase cleavage; (4) its extracellular domain (EpEX) acts as a ligand for EGFR and HGFR, activating AKT/MAPK signaling cascades that modulate PD-L1 stability, EMT, and metastasis; (5) it associates with integrin β1 to regulate FAK/ERK-dependent cell adhesion and migration; and (6) structural data show it forms cis-dimers but does not form the intercellular trans-oligomers originally proposed for homophilic adhesion, challenging the adhesion-molecule model."},"narrative":{"mechanistic_narrative":"EpCAM is a transmembrane epithelial glycoprotein that integrates cell-contact-dependent proliferative signaling with epithelial barrier maintenance [PMID:19136966, PMID:23486470]. Its extracellular domain adopts a characteristic disulfide architecture with a thyroglobulin type-1A region, and the full-length protein assembles into high-affinity non-covalent cis-dimers [PMID:11080501, PMID:11058587]. Upon cell-cell contact, EpCAM undergoes regulated intramembrane proteolysis: TACE sheds the EpEX ectodomain and presenilin-2 releases the intracellular domain EpICD, which translocates to the nucleus and forms a transcriptional complex with FHL2, beta-catenin, and Lef-1 to drive proliferative target genes including c-myc and cyclin D1, the latter feeding Rb phosphorylation and cell-cycle progression [PMID:19136966, PMID:19925656, PMID:22391566]. The shed EpEX ectodomain acts as a ligand for receptor tyrosine kinases: it binds EGFR to activate AKT/MAPK signaling that stabilizes PD-L1 and suppresses FOXO3a, and binds HGFR (c-Met) to cooperate with HGF in activating ERK and FAK-AKT signaling, stabilizing beta-catenin and Snail to promote EMT and metastasis [PMID:32978170, PMID:37543570]. In parallel, EpCAM physically associates with claudin-7 (and indirectly claudin-1), protecting these tight-junction proteins from lysosomal degradation; this stabilizing role is shared redundantly with TROP2 and is abolished by matriptase cleavage of EpCAM and TROP2 [PMID:23486470, PMID:32326212, PMID:35730316]. Through its cytoplasmic tail, a pseudosubstrate-like motif directly inhibits novel PKC, restraining Erk activity and myosin contractility to preserve cadherin-mediated adhesion and tissue integrity [PMID:24183651]. Loss of EpCAM function via exon-4 deletion disrupts the EpCAM/claudin-7 complex and increases intestinal permeability, the basis of congenital tufting enteropathy [PMID:24337010, PMID:25482158]. Structural and biophysical analyses establish that EpCAM forms cis-dimers but not intercellular trans-oligomers, arguing against a direct homophilic cell-cell adhesion function [PMID:30185875].","teleology":[{"year":1994,"claim":"Established that the amplifiability of the EpCAM/TROP1 locus is controlled by DNA methylation, providing the first regulatory handle on its genomic behavior in cancer cells.","evidence":"5-azacytidine demethylation, transfection, and Southern blot copy-number analysis in JAR choriocarcinoma cells","pmids":["8016075"],"confidence":"Medium","gaps":["Does not address the protein's function","Mechanism of methylation control at the endogenous locus not defined"]},{"year":2000,"claim":"Defined EpCAM's extracellular disulfide architecture, glycosylation sites, and showed it exists as non-covalent cis-dimers whose oligomeric state is required for cell aggregation activity, framing it as a putative adhesion molecule.","evidence":"MALDI-MS, peptide sequencing, sedimentation equilibrium, cross-linking, and cell aggregation inhibition assays on purified GA733-2/EpCAM","pmids":["11080501","11058587"],"confidence":"High","gaps":["Whether dimers act in cis or trans not resolved by these methods","No high-resolution structure of the intact protein"]},{"year":2003,"claim":"Connected loss of p53 to demethylation and amplification of the EpCAM/TROP1 gene, defining a p53→methylation→amplification axis relevant to tumorigenesis.","evidence":"wtTP53 reintroduction into p53-null cells with Southern blot methylation/copy-number analysis and in vitro Sss I methylation control","pmids":["12642870"],"confidence":"Medium","gaps":["Direct mechanism linking p53 to locus methylation unknown","Single lab"]},{"year":2009,"claim":"Resolved how EpCAM signals by demonstrating sequential TACE/presenilin-2 proteolysis releasing nuclear EpICD that complexes with FHL2/beta-catenin/Lef-1 to drive oncogenic transcription, and that this requires cell-cell contact.","evidence":"Protease inhibition, siRNA silencing, nuclear fractionation, xenograft oncogenesis, and density-dependent culture with pre-cleaved EpICD constructs","pmids":["19136966","19925656"],"confidence":"High","gaps":["Trigger coupling contact to TACE activity not molecularly defined","Identity of all EpICD target genes incomplete"]},{"year":2012,"claim":"Linked EpICD signaling to the cell cycle by showing FHL2-dependent cyclin D1 transcription, Rb phosphorylation, and induction of cyclins E and A.","evidence":"EpCAM knockdown/overexpression, cyclin RT-PCR/Western, FHL2 co-depletion rescue, and patient-tissue IHC","pmids":["22391566"],"confidence":"Medium","gaps":["Direct promoter occupancy by EpICD/FHL2 not shown","Single lab"]},{"year":2013,"claim":"Identified EpCAM's barrier function: it binds claudin-7 (and indirectly claudin-1) to protect them from lysosomal degradation, and validated this in a CTE mouse model and patient tissue.","evidence":"Reciprocal co-IP, knockdown with lysosomal-inhibitor rescue, TEER assays, and Epcam exon-4-deletion mouse with patient tissue confirmation","pmids":["23486470","24337010"],"confidence":"High","gaps":["Structural basis of EpCAM-claudin-7 interaction not defined","Paradoxical TJ accumulation on knockdown not fully explained"]},{"year":2013,"claim":"Revealed a contact-adhesion regulatory mechanism whereby EpCAM's cytoplasmic pseudosubstrate-like motif directly inhibits novel PKC to restrain Erk and myosin contractility and preserve tissue integrity.","evidence":"Xenopus loss-of-function, in vitro nPKC binding assays of the cytoplasmic tail, PKC/Erk/myosin readouts","pmids":["24183651"],"confidence":"High","gaps":["Relationship between PKC inhibition and RIP signaling not integrated","Which nPKC isoforms in mammalian epithelia not specified"]},{"year":2013,"claim":"Positioned EpCAM upstream of an NF-kB/RELA/IL-8 invasion axis in breast cancer through epistatic rescue.","evidence":"EpCAM siRNA knockdown, NF-kB reporter, RELA/IkBa Westerns, IL-8 ELISA, and RELA/IkBa rescue experiments","pmids":["23378578"],"confidence":"Medium","gaps":["Mechanism linking EpCAM to NF-kB activation unknown","Single lab and cell-type-specific"]},{"year":2014,"claim":"Quantified the barrier consequences of EpCAM loss, showing reduced electrical resistance, increased paracellular permeability, decreased ion transport, and reduced TJ protein expression.","evidence":"EpCAM shRNA in T84 cells and inducible Epcam exon-4-deletion mouse with TEER, permeability, ion transport, and Western readouts","pmids":["25482158"],"confidence":"High","gaps":["Causal chain from claudin loss to ion transport defects not dissected"]},{"year":2015,"claim":"Demonstrated that a defined-epitope antibody blocking EpICD production reduces nuclear signaling and tumor growth, validating RIP-dependent signaling as a therapeutic target.","evidence":"Antibody epitope mapping, EpICD Westerns, apoptosis assays, and xenograft models","pmids":["26317650"],"confidence":"Medium","gaps":["Mechanism by which epitope binding blocks cleavage unclear","Single lab"]},{"year":2018,"claim":"Challenged the homophilic-adhesion model by showing EpCAM forms cis-dimers but no detectable intercellular trans-oligomers.","evidence":"SAXS, cross-linking mass spectrometry, bead aggregation, and FLIM-FRET on cell surfaces","pmids":["30185875"],"confidence":"High","gaps":["Functional role of the cis-dimer not fully defined","Cannot exclude weak transient trans-interactions below detection"]},{"year":2020,"claim":"Established that the shed EpEX ectodomain acts as an EGFR ligand activating AKT/MAPK to stabilize PD-L1 and suppress FOXO3a, linking EpCAM to immune evasion.","evidence":"Co-IP of EpEX with EGFR, signaling Westerns, FOXO3a immunofluorescence, neutralizing antibody, and in vivo metastasis/orthotopic models","pmids":["32978170"],"confidence":"Medium","gaps":["Stoichiometry/affinity of EpEX-EGFR binding not quantified","Single lab"]},{"year":2020,"claim":"Showed matriptase cleaves EpCAM and TROP2 to destabilize claudins via lysosomal routing, with HAI proteins restraining this cleavage, defining a proteolytic switch on barrier integrity.","evidence":"In vitro cleavage with purified proteins, active-site mutant controls, HAI/matriptase knockdowns in HaCaT cells, and chloroquine rescue","pmids":["32326212"],"confidence":"High","gaps":["Physiological contexts where matriptase regulates EpCAM in vivo not established"]},{"year":2019,"claim":"Identified an EpCAM-integrin beta1 association controlling FAK/ERK-dependent adhesion and migration.","evidence":"Co-IP, CRISPR/Cas9 knockout, phospho-FAK/AKT/ERK Westerns, adhesion/migration/colony assays in CW-2 and A431 cells","pmids":["31806375"],"confidence":"Medium","gaps":["Direct vs indirect nature of EpCAM-integrin association unresolved","Single lab"]},{"year":2022,"claim":"Established genetic redundancy between EPCAM and TROP2 in claudin-7 stabilization and epithelial development using single and double knockout mice.","evidence":"Epcam, Trop2, and double-knockout mice with multi-tissue histology and claudin-7 immunofluorescence","pmids":["35730316"],"confidence":"High","gaps":["Molecular basis of functional substitution not defined","Tissue-specific differences in redundancy not fully mapped"]},{"year":2023,"claim":"Extended EpEX ligand activity to HGFR (c-Met), showing cooperation with HGF to drive ERK/FAK-AKT signaling, beta-catenin/Snail stabilization, EMT, and metastasis.","evidence":"IP, ELISA, FRET for EpEX-HGFR interaction, signaling Westerns, invasion assays, and tail-vein/orthotopic metastasis models","pmids":["37543570"],"confidence":"Medium","gaps":["Interplay between EpEX-EGFR and EpEX-HGFR signaling not reconciled","Single lab"]},{"year":null,"claim":"How EpCAM's distinct activities — RIP-driven nuclear transcription, EpEX-mediated RTK ligand function, claudin stabilization, integrin association, and PKC inhibition — are coordinated within a single cell and which dominate in specific tissue contexts remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model integrating proteolytic, ligand, and scaffolding functions","Context-dependent dominance of each mechanism unknown","No full-length structure linking dimerization to function"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[4]},{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[10,11]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[0,2]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[3,14]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3,4,7]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,10,11]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[2]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[8,15]},{"term_id":"R-HSA-1500931","term_label":"Cell-Cell communication","supporting_discovery_ids":[3,9]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[8]}],"complexes":["EpICD-FHL2-beta-catenin-Lef-1 transcriptional complex","EpCAM-claudin-7 complex"],"partners":["FHL2","CTNNB1","LEF1","CLDN7","CLDN1","EGFR","MET","ITGB1"],"other_free_text":[]}},"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":569,"is_preprint":false},{"pmid":"10606205","id":"PMC_10606205","title":"The biology of the 17-1A antigen (Ep-CAM).","date":"1999","source":"Journal of molecular medicine (Berlin, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/10606205","citation_count":467,"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":"17211480","id":"PMC_17211480","title":"EpCAM (CD326) finding its role in cancer.","date":"2007","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/17211480","citation_count":420,"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":268,"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":257,"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":236,"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":229,"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":158,"is_preprint":false},{"pmid":"22647938","id":"PMC_22647938","title":"EpCAM and its potential role in tumor-initiating cells.","date":"2012","source":"Cell adhesion & migration","url":"https://pubmed.ncbi.nlm.nih.gov/22647938","citation_count":152,"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":149,"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":146,"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":133,"is_preprint":false},{"pmid":"14508099","id":"PMC_14508099","title":"EpCAM: A new therapeutic target for an old cancer antigen.","date":"2003","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/14508099","citation_count":127,"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":"23264089","id":"PMC_23264089","title":"EPCAM deletion carriers constitute a unique subgroup of Lynch syndrome patients.","date":"2013","source":"Familial cancer","url":"https://pubmed.ncbi.nlm.nih.gov/23264089","citation_count":105,"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":"25477371","id":"PMC_25477371","title":"EpCAM and the biology of hepatic stem/progenitor cells.","date":"2014","source":"American journal of physiology. Gastrointestinal and liver physiology","url":"https://pubmed.ncbi.nlm.nih.gov/25477371","citation_count":98,"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":"17981779","id":"PMC_17981779","title":"EpCAM expression in normal, non-pathological tissues.","date":"2008","source":"Frontiers in bioscience : a journal and virtual library","url":"https://pubmed.ncbi.nlm.nih.gov/17981779","citation_count":89,"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":88,"is_preprint":false},{"pmid":"25103341","id":"PMC_25103341","title":"Dynamic EpCAM expression on circulating and disseminating tumor cells: causes and consequences.","date":"2014","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/25103341","citation_count":87,"is_preprint":false},{"pmid":"22482828","id":"PMC_22482828","title":"Significance of EpCAM and TROP2 expression in non-small cell lung cancer.","date":"2012","source":"World journal of surgical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/22482828","citation_count":86,"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":84,"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":"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":76,"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":75,"is_preprint":false},{"pmid":"11080501","id":"PMC_11080501","title":"Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM).","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11080501","citation_count":73,"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":"30479699","id":"PMC_30479699","title":"EpCAMhigh and EpCAMlow circulating tumor cells in metastatic prostate and breast cancer patients.","date":"2018","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/30479699","citation_count":68,"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":"32978170","id":"PMC_32978170","title":"EpCAM Signaling Promotes Tumor Progression and Protein Stability of PD-L1 through the EGFR Pathway.","date":"2020","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/32978170","citation_count":64,"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":59,"is_preprint":false},{"pmid":"19249674","id":"PMC_19249674","title":"EpCAM: another surface-to-nucleus missile.","date":"2009","source":"Cancer cell","url":"https://pubmed.ncbi.nlm.nih.gov/19249674","citation_count":58,"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":56,"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":"28403178","id":"PMC_28403178","title":"Relationship between epithelial cell adhesion molecule (EpCAM) overexpression and gastric cancer patients: A systematic review and meta-analysis.","date":"2017","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/28403178","citation_count":51,"is_preprint":false},{"pmid":"29329202","id":"PMC_29329202","title":"EpCAM Immunotherapy versus Specific Targeted Delivery of Drugs.","date":"2018","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/29329202","citation_count":50,"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":"11058587","id":"PMC_11058587","title":"Oligomeric state of the colon carcinoma-associated glycoprotein GA733-2 (Ep-CAM/EGP40) and its role in GA733-mediated homotypic cell-cell adhesion.","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11058587","citation_count":45,"is_preprint":false},{"pmid":"24465008","id":"PMC_24465008","title":"Expression of EpCAM(MF) and EpCAM(MT) variants in human carcinomas.","date":"2014","source":"Journal of clinical pathology","url":"https://pubmed.ncbi.nlm.nih.gov/24465008","citation_count":44,"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":"8016075","id":"PMC_8016075","title":"DNA methylation prevents the amplification of TROP1, a tumor-associated cell surface antigen gene.","date":"1994","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/8016075","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":"27842504","id":"PMC_27842504","title":"EpCAM as multi-tumour target for near-infrared fluorescence guided surgery.","date":"2016","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/27842504","citation_count":41,"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":"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":"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":"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":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":"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":36,"is_preprint":false},{"pmid":"24370904","id":"PMC_24370904","title":"EpCAM-targeted therapy for human hepatocellular carcinoma.","date":"2013","source":"Annals of surgical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/24370904","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":"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":33,"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":33,"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":"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":"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":31,"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":"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":30,"is_preprint":false},{"pmid":"12642870","id":"PMC_12642870","title":"Mutations of TP53 induce loss of DNA methylation and amplification of the TROP1 gene.","date":"2003","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/12642870","citation_count":30,"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":"31636732","id":"PMC_31636732","title":"EpCAMlow Circulating Tumor Cells: Gold in the Waste.","date":"2019","source":"Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/31636732","citation_count":29,"is_preprint":false},{"pmid":"28604994","id":"PMC_28604994","title":"EpCAM-expressing circulating tumor cells in colorectal cancer.","date":"2017","source":"The International journal of biological markers","url":"https://pubmed.ncbi.nlm.nih.gov/28604994","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":"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":28,"is_preprint":false},{"pmid":"27697766","id":"PMC_27697766","title":"EpCAM Inhibition Sensitizes Chemoresistant Leukemia to Immune Surveillance.","date":"2016","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/27697766","citation_count":28,"is_preprint":false},{"pmid":"17559145","id":"PMC_17559145","title":"Trop-1 are conserved growth stimulatory molecules that mark early stages of tumor progression.","date":"2007","source":"Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/17559145","citation_count":26,"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":26,"is_preprint":false},{"pmid":"35013776","id":"PMC_35013776","title":"Preclinical evaluation of [99mTc]Tc-labeled anti-EpCAM nanobody for EpCAM receptor expression imaging by immuno-SPECT/CT.","date":"2022","source":"European journal of nuclear medicine and molecular imaging","url":"https://pubmed.ncbi.nlm.nih.gov/35013776","citation_count":25,"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":"26401964","id":"PMC_26401964","title":"Expression of epithelial cell adhesion molecule (EpCAM) in oral squamous cell carcinoma.","date":"2015","source":"Histopathology","url":"https://pubmed.ncbi.nlm.nih.gov/26401964","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":"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":"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":23,"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":22,"is_preprint":false},{"pmid":"35682800","id":"PMC_35682800","title":"EpCAM- and EGFR-Specific Antibody Drug Conjugates for Triple-Negative Breast Cancer Treatment.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35682800","citation_count":22,"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":"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":"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":20,"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":"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":"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":"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":"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":"35730316","id":"PMC_35730316","title":"EPCAM and TROP2 share a role in claudin stabilization and development of intestinal and extraintestinal epithelia in mice.","date":"2022","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/35730316","citation_count":17,"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}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":48450,"output_tokens":5772,"usd":0.115965,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14273,"output_tokens":5190,"usd":0.100557,"stage2_stop_reason":"end_turn"},"total_usd":0.216522,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2009,\n      \"finding\": \"EpCAM undergoes regulated intramembrane proteolysis (RIP): its ectodomain EpEX is shed by TACE, and the remaining stub is cleaved by presenilin-2 to release the intracellular domain EpICD, which translocates to the nucleus. Nuclear EpICD forms a complex with FHL2, β-catenin, and Lef-1 that binds Lef-1 consensus DNA sites, drives target gene transcription, and is oncogenic in immunodeficient mice. Pharmacological or genetic inhibition of either protease impairs EpCAM-dependent proliferative signaling.\",\n      \"method\": \"Pharmacological inhibition (TACE/presenilin-2 inhibitors), genetic silencing (siRNA), conditional cell systems, confocal microscopy, immunoblotting, nuclear fractionation, xenograft oncogenesis assay, patient tissue immunohistochemistry\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (biochemical cleavage assays, genetic silencing of both proteases, nuclear fractionation, in vivo oncogenesis, patient tissue validation) in a single rigorous study; widely replicated by subsequent papers\",\n      \"pmids\": [\"19136966\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Activation of EpCAM's RIP-dependent oncogenic signaling requires cell-to-cell contact: when intercellular contact is prevented, EpCAM does not confer growth advantage. Contact triggers initial cleavage (juxtacrine), releasing soluble EpEX that can act in a paracrine manner. The pre-cleaved EpICD fragment bypasses the contact requirement but still requires nuclear translocation to induce c-myc and proliferation.\",\n      \"method\": \"Density-dependent cell culture experiments, conditional cell systems expressing pre-cleaved EpICD, confocal laser scanning microscopy, immunoblotting, cell counting, c-myc reporter assays\",\n      \"journal\": \"BMC cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple complementary cell-based assays in a single lab; consistent with the RIP mechanism established in PMID 19136966\",\n      \"pmids\": [\"19925656\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"EpCAM drives cell cycle progression by upregulating cyclin D1 transcription in an FHL2-dependent manner. Downstream consequences include phosphorylation of retinoblastoma protein (Rb) and induction of cyclins E and A. In vivo, EpCAM expression level positively correlates with Ki67, nuclear cyclin D1, and Rb phosphorylation.\",\n      \"method\": \"EpCAM knockdown/overexpression in cancer cells, RT-PCR and Western blotting for cyclins, Rb phosphorylation assays, FHL2 co-depletion rescue experiments, immunohistochemistry of patient tissues\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic manipulation with defined molecular readouts, corroborated by in vivo IHC, single lab\",\n      \"pmids\": [\"22391566\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EpCAM physically associates with claudin-7 (tight interaction) and, indirectly via claudin-7, with claudin-1, but not claudin-2 or claudin-4. This interaction stabilizes claudin-7 and claudin-1 by preventing their lysosomal degradation. EpCAM knockdown reduces claudin-7 and claudin-1 protein levels, which are restored by lysosomal inhibitors; this also paradoxically increases claudin accumulation at tight junctions and enhances transepithelial electroresistance.\",\n      \"method\": \"shRNA knockdown of EpCAM in T84 and Caco-2 cells, preparative immunoprecipitation, co-immunoprecipitation, co-transfection experiments, lysosomal inhibitor rescue, immunofluorescence microscopy, transepithelial electrical resistance measurement\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — reciprocal co-IP, preparative IP, genetic knockdown, lysosomal rescue, functional TJ assays; multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"23486470\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EpCAM acts as a potent inhibitor of novel protein kinase C (nPKC) through a short pseudosubstrate-like motif in its cytoplasmic tail that binds nPKCs with high affinity. Loss of EpCAM in amphibian embryos sequentially overstimulates PKC, activates the Erk pathway, exacerbates myosin contractility, disrupts cadherin-mediated adhesion, and causes tissue dissociation. This PKC-inhibitory mechanism is shared with other plasma membrane adhesion molecules.\",\n      \"method\": \"Loss-of-function in Xenopus embryos, PKC activity assays, Erk pathway measurements, myosin contractility readouts, in vitro binding assays of EpCAM cytoplasmic tail segment to nPKCs, bioinformatics motif analysis\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro binding assay identifying the minimal inhibitory motif, loss-of-function in intact tissue with sequential phenotypic readouts, multiple orthogonal methods\",\n      \"pmids\": [\"24183651\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"GA733-2/EpCAM extracellular domain disulfide-bonding pattern was determined: cysteines 1–6 form a novel pattern (Cys1-Cys4, Cys2-Cys6, Cys3-Cys5) distinct from EGF-like structure, and cysteines 7–12 form a thyroglobulin type 1A pattern (Cys7-Cys8, Cys9-Cys10, Cys11-Cys12). N-linked glycosylation occurs at Asn88 (complete) and Asn51 (partial); Asn175 is not glycosylated; no O-linked carbohydrate was detected.\",\n      \"method\": \"MALDI mass spectrometry, N-terminal sequencing of tryptic peptides, chemical reduction and alkylation, glycosylation site analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct biochemical structural determination with mass spectrometry and sequencing; constitutes the primary structural characterization of EpCAM's disulfide architecture\",\n      \"pmids\": [\"11080501\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Full-length GA733-2/EpCAM exists as high-affinity non-covalent cis-dimers (Kd <10 nM for monomer-dimer) and lower-affinity tetramers (~10 µM for dimer-tetramer) in solution. The extracellular domain alone is monomeric. Cell-cell adhesion activity resides in the full-length dimer/tetramer; monomeric extracellular domain is inactive in cell aggregation inhibition assays.\",\n      \"method\": \"Sedimentation equilibrium ultracentrifugation, chemical cross-linking of purified protein and cells in suspension vs. monolayer, cell aggregation inhibition assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — biophysical (sedimentation equilibrium) plus biochemical (cross-linking) plus functional (aggregation) methods establish oligomeric state and link it to adhesion activity\",\n      \"pmids\": [\"11058587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EpCAM monomers do not associate into intercellular homo-oligomers capable of mediating cell-cell adhesion. EpCAM forms stable cis-dimers on the surface of cells with pre-formed contacts (detected by FLIM-FRET), but no trans-oligomers between opposing cells were detectable. These findings argue against EpCAM functioning as a direct homophilic cell-cell adhesion molecule.\",\n      \"method\": \"SAXS, cross-linking mass spectrometry (XL-MS), bead aggregation assays, FLIM-FRET on cell surfaces\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — multiple structural/biophysical methods (SAXS, XL-MS, FLIM-FRET) in one study; result is explicitly a negative finding regarding intercellular homo-oligomerization\",\n      \"pmids\": [\"30185875\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"In a mouse model of congenital tufting enteropathy (CTE), deletion of EpCAM exon 4 causes mislocalization of residual mutant EpCAM protein, loss of claudin-7 colocalization, and disruption of the EpCAM/claudin-7 complex, leading to enhanced intestinal permeability and epithelial cell migration. These findings were confirmed in CTE patient intestinal tissue.\",\n      \"method\": \"Cre-LoxP mouse model (Epcam Δ4/Δ4), histology, light and electron microscopy, immunohistochemistry, permeability assays, co-localization studies in mouse and patient tissue\",\n      \"journal\": \"American journal of physiology. Gastrointestinal and liver physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo genetic model validated against human patient tissue, with functional permeability/migration readouts and protein complex analysis\",\n      \"pmids\": [\"24337010\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"EpCAM mutation (exon 4 deletion) in intestinal epithelial cells leads to decreased electrical resistance, increased paracellular permeability, and decreased ion transport. In the inducible mouse model, EpCAM deficiency also decreases tight junctional protein expression in intestine.\",\n      \"method\": \"EpCAM shRNA knockdown in T84 colonic epithelial cells, transepithelial electrical resistance measurement, permeability assays, ion transport assays, tamoxifen-inducible EpCAM Δ4/Δ4 mouse model, Western blotting\",\n      \"journal\": \"Journal of molecular medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — both in vitro (knockdown) and in vivo (inducible mouse model) with quantitative functional barrier and transport readouts\",\n      \"pmids\": [\"25482158\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"The EGF-like domain I within the extracellular domain of EpCAM (EpEX) binds EGFR, activating AKT and MAPK signaling. AKT signaling inhibits FOXO3a and stabilizes PD-L1 protein; MAPK signaling also contributes to PD-L1 stabilization. Neutralizing EpCAM (anti-EpCAM antibody EpAb2-6) reduces PD-L1 levels and promotes FOXO3a nuclear translocation and HtrA2-mediated apoptosis.\",\n      \"method\": \"Co-immunoprecipitation of EpEX with EGFR, Western blotting for AKT/MAPK/FOXO3a/PD-L1, immunofluorescence for FOXO3a localization, in vivo mouse metastasis and orthotopic colorectal cancer models\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP establishing EpEX-EGFR interaction combined with signaling pathway readouts and in vivo validation, single lab\",\n      \"pmids\": [\"32978170\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The extracellular domain of EpCAM (EpEX) binds HGFR (c-Met) and cooperates with HGF to activate downstream ERK and FAK-AKT signaling, stabilizes active β-catenin and Snail by reducing GSK3β activity, and promotes EMT and metastasis of colon cancer cells.\",\n      \"method\": \"Immunoprecipitation, ELISA, FRET to confirm EpEX-HGFR interaction, Western blotting for downstream signaling, cell proliferation/migration/invasion assays, tail-vein metastasis and orthotopic animal models\",\n      \"journal\": \"Journal of translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — IP/FRET establishing the EpEX-HGFR interaction plus downstream signaling and in vivo validation, single lab\",\n      \"pmids\": [\"37543570\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EpCAM knockdown in breast cancer cells decreases NF-κB transcription factor activity (reduced RELA phosphorylation, increased IκBα) and reduces IL-8 expression. Rescue experiments show that RELA ablation or forced IκBα expression prevented EpCAM-dependent IL-8 promoter reactivation, placing EpCAM upstream of the NF-κB/RELA/IL-8 axis in breast cancer invasion.\",\n      \"method\": \"EpCAM siRNA knockdown in breast cancer cell lines, NF-κB reporter assays, RELA phosphorylation Western blotting, IκBα Western blotting, IL-8 ELISA, IL-1β stimulation, functional rescue with RELA siRNA and IκBα overexpression\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockdown with epistatic rescue experiments defining pathway position, single lab\",\n      \"pmids\": [\"23378578\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"EpCAM associates with integrin β1 (demonstrated by co-immunoprecipitation). EpCAM knockout (CRISPR/Cas9) in colorectal CW-2 and epidermoid A431 cells reduces integrin α5 expression, decreases FAK, AKT, and ERK phosphorylation, and impairs cell adhesion, migration, and colony formation.\",\n      \"method\": \"CRISPR/Cas9 knockout, co-immunoprecipitation, Western blotting for phospho-FAK/AKT/ERK, cell adhesion assays, migration assays, colony formation assays\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — co-IP identifying EpCAM–integrin β1 association plus CRISPR KO with defined signaling readouts, single lab\",\n      \"pmids\": [\"31806375\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Matriptase cleaves both EpCAM and TROP2 in keratinocytes (HaCaT cells). Matriptase cleavage of EpCAM and TROP2 destabilizes claudin-1 and claudin-7 by routing them to lysosomes. This cleavage is inhibited by HAI proteins (HAI-1 more than HAI-2); dual knockdown of HAI-1 and HAI-2 causes near-complete cleavage of EpCAM/TROP2 and drastic reduction of claudins, reversible by concurrent matriptase knockdown. Protease-disabled matriptase or G827R mutant does not cleave EpCAM or TROP2.\",\n      \"method\": \"In vitro cleavage assay with purified recombinant proteins, co-transfection of matriptase in 293T cells, siRNA knockdown of HAI-1/HAI-2/matriptase in HaCaT cells, Western blotting, lysosomal inhibitor (chloroquine) rescue\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro reconstitution with purified proteins, active-site mutant controls, cell-based corroboration with epistatic knockdowns, multiple orthogonal methods\",\n      \"pmids\": [\"32326212\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"EPCAM and TROP2 share redundant roles in stabilizing claudin-7 at the cell membrane and in epithelial development across multiple tissues. TROP2 can compensate for loss of EPCAM in claudin-7 stabilization in tissues co-expressing both proteins; combined loss causes much more severe developmental defects than either single knockout alone.\",\n      \"method\": \"Epcam knockout mice, Trop2 knockout mice, Epcam/Trop2 double knockout mice, histology of multiple tissues, immunofluorescence for claudin-7 localization and expression, viability and growth tracking\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis using single and double knockout mice with molecular readout (claudin-7 stabilization), replicated across multiple tissues\",\n      \"pmids\": [\"35730316\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"DNA methylation of the TROP1/EpCAM gene locus prevents gene amplification after transfection. Demethylation of TROP1 (by 5-azacytidine treatment) allows efficient amplification of transfected TROP1 copies (up to 40 copies per haploid genome), demonstrating that methylation status controls the amplifiability of this locus.\",\n      \"method\": \"5-azacytidine demethylation treatment of JAR choriocarcinoma cells, DNA transfection, FACS-based selection, Southern blot copy-number analysis, in vitro Sss I methylase treatment of DNA\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — enzymatic methylation rescue experiment plus demethylation treatment establishing causal link between methylation and amplification resistance, single lab\",\n      \"pmids\": [\"8016075\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Loss of TP53 (p53) induces demethylation of the TROP1/EpCAM gene, leading to TROP1 gene amplification. Reintroduction of wild-type TP53 into p53-null cells reverses the demethylation. In vitro methylation of transfected TROP1 DNA with Sss I methylase prevents amplification, establishing a p53→DNA methylation→TROP1 amplification axis.\",\n      \"method\": \"TP53 transfection into TP53-null cells, Southern blot for methylation and copy number, in vitro methylation with Sss I methylase, gene amplification assay in vivo\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic rescue (wtTP53 reintroduction) plus in vitro methylation control, single lab\",\n      \"pmids\": [\"12642870\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"An anti-EpCAM antibody (EpAb2-6) binding to positions Y95 and D96 of the EGF-II/TY domain of EpCAM inhibits production of EpICD, thereby decreasing its nuclear translocation and downstream signal activation (oncogenic signaling). This antibody induces cancer cell apoptosis in vitro and inhibits tumor growth in vivo.\",\n      \"method\": \"Antibody epitope mapping, Western blotting for EpICD production and nuclear translocation, cell apoptosis assays, xenograft mouse models\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epitope-defined antibody blocking EpICD generation with downstream signaling readouts, single lab\",\n      \"pmids\": [\"26317650\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EpCAM is a transmembrane glycoprotein that functions through multiple mechanisms: (1) it undergoes regulated intramembrane proteolysis sequentially by TACE and presenilin-2, releasing soluble EpEX and nuclear EpICD, which complexes with FHL2, β-catenin, and Lef-1 to drive proliferative gene transcription (including cyclin D1 and c-myc) in a cell-contact-initiated manner; (2) its cytoplasmic pseudosubstrate-like motif directly inhibits novel PKC, suppressing actomyosin contractility and promoting tissue plasticity; (3) it stabilizes claudin-1 and claudin-7 by protecting them from lysosomal degradation via physical association—a function shared redundantly with TROP2 and disrupted by matriptase cleavage; (4) its extracellular domain (EpEX) acts as a ligand for EGFR and HGFR, activating AKT/MAPK signaling cascades that modulate PD-L1 stability, EMT, and metastasis; (5) it associates with integrin β1 to regulate FAK/ERK-dependent cell adhesion and migration; and (6) structural data show it forms cis-dimers but does not form the intercellular trans-oligomers originally proposed for homophilic adhesion, challenging the adhesion-molecule model.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EpCAM is a transmembrane epithelial glycoprotein that integrates cell-contact-dependent proliferative signaling with epithelial barrier maintenance [#0, #3]. Its extracellular domain adopts a characteristic disulfide architecture with a thyroglobulin type-1A region, and the full-length protein assembles into high-affinity non-covalent cis-dimers [#5, #6]. Upon cell-cell contact, EpCAM undergoes regulated intramembrane proteolysis: TACE sheds the EpEX ectodomain and presenilin-2 releases the intracellular domain EpICD, which translocates to the nucleus and forms a transcriptional complex with FHL2, beta-catenin, and Lef-1 to drive proliferative target genes including c-myc and cyclin D1, the latter feeding Rb phosphorylation and cell-cycle progression [#0, #1, #2]. The shed EpEX ectodomain acts as a ligand for receptor tyrosine kinases: it binds EGFR to activate AKT/MAPK signaling that stabilizes PD-L1 and suppresses FOXO3a, and binds HGFR (c-Met) to cooperate with HGF in activating ERK and FAK-AKT signaling, stabilizing beta-catenin and Snail to promote EMT and metastasis [#10, #11]. In parallel, EpCAM physically associates with claudin-7 (and indirectly claudin-1), protecting these tight-junction proteins from lysosomal degradation; this stabilizing role is shared redundantly with TROP2 and is abolished by matriptase cleavage of EpCAM and TROP2 [#3, #14, #15]. Through its cytoplasmic tail, a pseudosubstrate-like motif directly inhibits novel PKC, restraining Erk activity and myosin contractility to preserve cadherin-mediated adhesion and tissue integrity [#4]. Loss of EpCAM function via exon-4 deletion disrupts the EpCAM/claudin-7 complex and increases intestinal permeability, the basis of congenital tufting enteropathy [#8, #9]. Structural and biophysical analyses establish that EpCAM forms cis-dimers but not intercellular trans-oligomers, arguing against a direct homophilic cell-cell adhesion function [#7].\",\n  \"teleology\": [\n    {\n      \"year\": 1994,\n      \"claim\": \"Established that the amplifiability of the EpCAM/TROP1 locus is controlled by DNA methylation, providing the first regulatory handle on its genomic behavior in cancer cells.\",\n      \"evidence\": \"5-azacytidine demethylation, transfection, and Southern blot copy-number analysis in JAR choriocarcinoma cells\",\n      \"pmids\": [\"8016075\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not address the protein's function\", \"Mechanism of methylation control at the endogenous locus not defined\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Defined EpCAM's extracellular disulfide architecture, glycosylation sites, and showed it exists as non-covalent cis-dimers whose oligomeric state is required for cell aggregation activity, framing it as a putative adhesion molecule.\",\n      \"evidence\": \"MALDI-MS, peptide sequencing, sedimentation equilibrium, cross-linking, and cell aggregation inhibition assays on purified GA733-2/EpCAM\",\n      \"pmids\": [\"11080501\", \"11058587\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether dimers act in cis or trans not resolved by these methods\", \"No high-resolution structure of the intact protein\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Connected loss of p53 to demethylation and amplification of the EpCAM/TROP1 gene, defining a p53\\u2192methylation\\u2192amplification axis relevant to tumorigenesis.\",\n      \"evidence\": \"wtTP53 reintroduction into p53-null cells with Southern blot methylation/copy-number analysis and in vitro Sss I methylation control\",\n      \"pmids\": [\"12642870\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct mechanism linking p53 to locus methylation unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Resolved how EpCAM signals by demonstrating sequential TACE/presenilin-2 proteolysis releasing nuclear EpICD that complexes with FHL2/beta-catenin/Lef-1 to drive oncogenic transcription, and that this requires cell-cell contact.\",\n      \"evidence\": \"Protease inhibition, siRNA silencing, nuclear fractionation, xenograft oncogenesis, and density-dependent culture with pre-cleaved EpICD constructs\",\n      \"pmids\": [\"19136966\", \"19925656\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Trigger coupling contact to TACE activity not molecularly defined\", \"Identity of all EpICD target genes incomplete\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Linked EpICD signaling to the cell cycle by showing FHL2-dependent cyclin D1 transcription, Rb phosphorylation, and induction of cyclins E and A.\",\n      \"evidence\": \"EpCAM knockdown/overexpression, cyclin RT-PCR/Western, FHL2 co-depletion rescue, and patient-tissue IHC\",\n      \"pmids\": [\"22391566\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct promoter occupancy by EpICD/FHL2 not shown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identified EpCAM's barrier function: it binds claudin-7 (and indirectly claudin-1) to protect them from lysosomal degradation, and validated this in a CTE mouse model and patient tissue.\",\n      \"evidence\": \"Reciprocal co-IP, knockdown with lysosomal-inhibitor rescue, TEER assays, and Epcam exon-4-deletion mouse with patient tissue confirmation\",\n      \"pmids\": [\"23486470\", \"24337010\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of EpCAM-claudin-7 interaction not defined\", \"Paradoxical TJ accumulation on knockdown not fully explained\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Revealed a contact-adhesion regulatory mechanism whereby EpCAM's cytoplasmic pseudosubstrate-like motif directly inhibits novel PKC to restrain Erk and myosin contractility and preserve tissue integrity.\",\n      \"evidence\": \"Xenopus loss-of-function, in vitro nPKC binding assays of the cytoplasmic tail, PKC/Erk/myosin readouts\",\n      \"pmids\": [\"24183651\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relationship between PKC inhibition and RIP signaling not integrated\", \"Which nPKC isoforms in mammalian epithelia not specified\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Positioned EpCAM upstream of an NF-kB/RELA/IL-8 invasion axis in breast cancer through epistatic rescue.\",\n      \"evidence\": \"EpCAM siRNA knockdown, NF-kB reporter, RELA/IkBa Westerns, IL-8 ELISA, and RELA/IkBa rescue experiments\",\n      \"pmids\": [\"23378578\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking EpCAM to NF-kB activation unknown\", \"Single lab and cell-type-specific\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Quantified the barrier consequences of EpCAM loss, showing reduced electrical resistance, increased paracellular permeability, decreased ion transport, and reduced TJ protein expression.\",\n      \"evidence\": \"EpCAM shRNA in T84 cells and inducible Epcam exon-4-deletion mouse with TEER, permeability, ion transport, and Western readouts\",\n      \"pmids\": [\"25482158\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Causal chain from claudin loss to ion transport defects not dissected\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Demonstrated that a defined-epitope antibody blocking EpICD production reduces nuclear signaling and tumor growth, validating RIP-dependent signaling as a therapeutic target.\",\n      \"evidence\": \"Antibody epitope mapping, EpICD Westerns, apoptosis assays, and xenograft models\",\n      \"pmids\": [\"26317650\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which epitope binding blocks cleavage unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Challenged the homophilic-adhesion model by showing EpCAM forms cis-dimers but no detectable intercellular trans-oligomers.\",\n      \"evidence\": \"SAXS, cross-linking mass spectrometry, bead aggregation, and FLIM-FRET on cell surfaces\",\n      \"pmids\": [\"30185875\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional role of the cis-dimer not fully defined\", \"Cannot exclude weak transient trans-interactions below detection\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established that the shed EpEX ectodomain acts as an EGFR ligand activating AKT/MAPK to stabilize PD-L1 and suppress FOXO3a, linking EpCAM to immune evasion.\",\n      \"evidence\": \"Co-IP of EpEX with EGFR, signaling Westerns, FOXO3a immunofluorescence, neutralizing antibody, and in vivo metastasis/orthotopic models\",\n      \"pmids\": [\"32978170\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Stoichiometry/affinity of EpEX-EGFR binding not quantified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed matriptase cleaves EpCAM and TROP2 to destabilize claudins via lysosomal routing, with HAI proteins restraining this cleavage, defining a proteolytic switch on barrier integrity.\",\n      \"evidence\": \"In vitro cleavage with purified proteins, active-site mutant controls, HAI/matriptase knockdowns in HaCaT cells, and chloroquine rescue\",\n      \"pmids\": [\"32326212\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological contexts where matriptase regulates EpCAM in vivo not established\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified an EpCAM-integrin beta1 association controlling FAK/ERK-dependent adhesion and migration.\",\n      \"evidence\": \"Co-IP, CRISPR/Cas9 knockout, phospho-FAK/AKT/ERK Westerns, adhesion/migration/colony assays in CW-2 and A431 cells\",\n      \"pmids\": [\"31806375\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect nature of EpCAM-integrin association unresolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Established genetic redundancy between EPCAM and TROP2 in claudin-7 stabilization and epithelial development using single and double knockout mice.\",\n      \"evidence\": \"Epcam, Trop2, and double-knockout mice with multi-tissue histology and claudin-7 immunofluorescence\",\n      \"pmids\": [\"35730316\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of functional substitution not defined\", \"Tissue-specific differences in redundancy not fully mapped\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended EpEX ligand activity to HGFR (c-Met), showing cooperation with HGF to drive ERK/FAK-AKT signaling, beta-catenin/Snail stabilization, EMT, and metastasis.\",\n      \"evidence\": \"IP, ELISA, FRET for EpEX-HGFR interaction, signaling Westerns, invasion assays, and tail-vein/orthotopic metastasis models\",\n      \"pmids\": [\"37543570\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interplay between EpEX-EGFR and EpEX-HGFR signaling not reconciled\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How EpCAM's distinct activities — RIP-driven nuclear transcription, EpEX-mediated RTK ligand function, claudin stabilization, integrin association, and PKC inhibition — are coordinated within a single cell and which dominate in specific tissue contexts remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model integrating proteolytic, ligand, and scaffolding functions\", \"Context-dependent dominance of each mechanism unknown\", \"No full-length structure linking dimerization to function\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [10, 11]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [3, 14]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3, 4, 7]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 10, 11]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [8, 15]},\n      {\"term_id\": \"R-HSA-1500931\", \"supporting_discovery_ids\": [3, 9]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"complexes\": [\n      \"EpICD-FHL2-beta-catenin-Lef-1 transcriptional complex\",\n      \"EpCAM-claudin-7 complex\"\n    ],\n    \"partners\": [\n      \"FHL2\",\n      \"CTNNB1\",\n      \"LEF1\",\n      \"CLDN7\",\n      \"CLDN1\",\n      \"EGFR\",\n      \"MET\",\n      \"ITGB1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}