{"gene":"MUC16","run_date":"2026-04-29T11:37:56","timeline":{"discoveries":[{"year":2003,"finding":"CA125/MUC16 binds to mesothelin, a GPI-anchored cell surface molecule, mediating heterotypic cell adhesion between ovarian cancer cells and mesothelial cells; anti-mesothelin antibody blocks this binding and cell attachment.","method":"Expression cloning, flow cytometry, immunoprecipitation, cell adhesion assay with antibody blockade","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP and functional adhesion assay with antibody blockade, replicated in multiple subsequent studies","pmids":["14676194"],"is_preprint":false},{"year":2008,"finding":"MUC16 cytoplasmic tail binds the N-terminus of ERM (ezrin/radixin/moesin) actin-binding proteins, linking MUC16 to the actin cytoskeleton; MUC16 knockdown in corneal epithelial cells reduces barrier function (increased dye penetrance) and increases Staphylococcus aureus binding.","method":"Cytoplasmic tail peptide pull-down of ERM proteins, RNA interference knockdown, rose bengal dye exclusion assay, bacterial binding assay, scanning and immunoelectron microscopy","journal":"Investigative ophthalmology & visual science","confidence":"High","confidence_rationale":"Tier 2 — direct peptide pull-down plus functional RNAi phenotype with multiple orthogonal readouts","pmids":["17898272"],"is_preprint":false},{"year":2008,"finding":"A 64-amino acid region (residues 296–359) at the N-terminal of cell-surface mesothelin is the minimum fragment sufficient for binding CA125/MUC16; tyrosine 318 substitution with alanine abolishes CA125 binding, and partial loss is caused by mutations at tryptophan 321 and glutamic acid 324.","method":"Truncated mutagenesis, alanine replacement, Western blot overlay assay, ELISA, flow cytometry on cancer cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — in vitro mutagenesis with quantitative binding assays and cell-based validation","pmids":["19075018"],"is_preprint":false},{"year":2003,"finding":"CA125/MUC16 is a counter receptor for galectin-1; soluble and membrane-associated CA125 fragments bind galectin-1 specifically via beta-galactose-terminated O-linked oligosaccharide chains, with preference for galectin-1 over galectin-3, and CA125 expression enhances galectin-1 presentation on the cell surface.","method":"Mass spectrometry, immunological analysis, binding specificity assays with glycan inhibitors, FACS-based non-conventional secretion reconstitution assay","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1 — MS identification plus mechanistic glycan-dependent binding assays with functional reconstitution","pmids":["12615972"],"is_preprint":false},{"year":2007,"finding":"MUC16 is lost from uterodome (pinopode) surfaces during the receptive phase of the uterine cycle (LH+6 to LH+8); siRNA knockdown of MUC16 (but not MUC1) in ECC-1 uterine epithelial cells increased trophoblast cell adhesion, demonstrating MUC16 acts as a barrier to trophoblast adherence.","method":"Immunofluorescence microscopy on uterine biopsies, siRNA knockdown, trophoblast adhesion assay","journal":"Biology of reproduction","confidence":"High","confidence_rationale":"Tier 2 — specific siRNA with functional adhesion readout and parallel MUC1 control showing specificity","pmids":["17942799"],"is_preprint":false},{"year":2014,"finding":"MUC16 knockdown in epithelial cells decreases all barrier functions tested (dye penetrance, bacterial adherence and invasion, transepithelial resistance, tight junction formation, and apical surface size), while MUC1 knockdown increases barrier to dye and bacteria; demonstrating MUC16 provides the dominant epithelial barrier.","method":"Stable shRNA knockdown of MUC16 and MUC1 independently, dye penetrance assay, bacterial invasion assay, transepithelial resistance measurement, tight junction immunostaining","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function with multiple orthogonal barrier readouts and internal control (MUC1 KD)","pmids":["24968021"],"is_preprint":false},{"year":2014,"finding":"MT1-MMP (MMP-14) mediates ectodomain shedding of MUC16/CA-125; overexpression of catalytically active MT1-MMP in OVCA433 cells causes loss of surface MUC16, while inactive E240A mutant does not; MUC16 shedding reduces adhesion to peritoneal tissue explants but enhances meso-mimetic invasion.","method":"MT1-MMP overexpression and catalytic mutant (E240A) experiments, surface MUC16 immunoreactivity, 3D meso-mimetic culture adhesion assay, ex vivo peritoneal explant adhesion assay, invasion assay","journal":"Biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — catalytic mutant controls plus multiple functional readouts in physiologically relevant model","pmids":["25205731"],"is_preprint":false},{"year":2016,"finding":"The C-terminal fragment of MUC16 (MUC16C) interacts with β-catenin (requiring β-catenin's trans-activation domain); this interaction facilitates cytosol-to-nucleus translocation of β-catenin, activating Wnt/β-catenin signaling to promote cell proliferation and migration, and tumorigenesis in nude mice.","method":"Co-immunoprecipitation with endogenous proteins using self-made MUC16 monoclonal antibody, domain-mapping, nuclear fractionation, reporter assays, MUC16 knockdown, xenograft tumor model","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP with domain mapping and in vivo validation, single lab","pmids":["27167110"],"is_preprint":false},{"year":2017,"finding":"The C-terminal domain of MUC16 activates JAK2/STAT3/glucocorticoid receptor (GR) signaling to upregulate TSPYL5 in lung cancer cells; STAT3 inhibition reduces GR and TSPYL5; MUC16 overexpression induces cisplatin and gemcitabine resistance by downregulating p53.","method":"Stable shRNA knockdown and MUC16-Cter overexpression, transcriptome analysis, rescue experiments, STAT3 inhibitor treatment, Western blot of signaling proteins, in vivo tumor growth assay","journal":"Clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — KD/OE with pathway rescue and in vivo validation, single lab","pmids":["28196872"],"is_preprint":false},{"year":2015,"finding":"MUC16 knockdown in pancreatic cancer cells reduces mTOR activity and c-MYC expression; ectopic c-MYC restores the metabolic and physiological alterations caused by MUC16 knockdown, including reduced glucose uptake, lactate secretion, and motility. Metabolomics showed MUC16 promotes glycolytic and nucleotide metabolite pools.","method":"shRNA knockdown, c-MYC overexpression rescue, glucose uptake and lactate secretion assays, LC-MS/MS metabolomics, Western blot for mTOR activity","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 — metabolomics plus genetic rescue, single lab","pmids":["26046375"],"is_preprint":false},{"year":2017,"finding":"Oncogenic KRAS upregulates MUC16 expression and CA125 shedding via the ERK/c-Myc axis in pancreatic cancer; c-Myc binds to the MUC16 promoter to transcriptionally activate its expression.","method":"KRAS manipulation in vitro and in vivo, ERK inhibitor treatment, c-Myc ChIP on MUC16 promoter, correlation analyses","journal":"Molecular cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — promoter ChIP plus pathway epistasis, single lab","pmids":["28108627"],"is_preprint":false},{"year":2012,"finding":"MUC16 and mesothelin are co-expressed and physically interact (co-immunoprecipitation) in pancreatic ductal adenocarcinoma; MUC16 shRNA knockdown and antibody blockade of MUC16-mesothelin binding inhibit invasion and migration of pancreatic cancer cells.","method":"Microarray, immunohistochemistry, co-immunoprecipitation from pancreatic cancer tissue, shRNA knockdown, antibody blockade, invasion and migration assays","journal":"Cancer science","confidence":"High","confidence_rationale":"Tier 2 — Co-IP from native tissue plus functional knockdown and antibody blockade with invasion readout","pmids":["22320398"],"is_preprint":false},{"year":2019,"finding":"MUC16 suppresses human and murine NK cell cytolysis and NK-tumor conjugate formation; MUC16-knockdown OVCAR-3 cells show increased susceptibility to NK and macrophage killing; mice bearing MUC16-knockdown tumors show >2-fold increased survival. MUC16 also suppresses ADCC by murine splenocytes.","method":"Target cell cytolysis assays, doublet formation assays, in vivo mouse survival model with MUC16-knockdown cells, in vitro NK/macrophage cytotoxicity assays","journal":"Gynecologic oncology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal immune cytotoxicity assays and in vivo model, replicates human and murine systems","pmids":["30626487"],"is_preprint":false},{"year":2017,"finding":"MUC16 oncogenic signaling through its C-terminal ectodomain requires MGAT5-dependent N-glycosylation at two specific asparagine sites; oncogenic effects depend on Galectin-3 and growth factor receptors colocalized on lipid rafts; N-glycosylation site-directed antibodies block Galectin-3-mediated MUC16 interactions and inhibit ovarian cancer cell invasion and in vivo xenograft growth.","method":"MGAT5 loss-of-function, synthetic MUC16 glycopeptide-based monoclonal antibodies, galectin-3 expression knockdown, lipid raft fractionation, in vitro invasion assay, in vivo xenograft model","journal":"ACS chemical biology","confidence":"High","confidence_rationale":"Tier 1 — glycan-specific mutagenesis/ablation plus in vitro and in vivo functional validation with mechanistic antibodies","pmids":["28617578"],"is_preprint":false},{"year":2020,"finding":"MUC16 enhances pancreatic cancer tumor malignancy through activation of AKT and GSK3β oncogenic signaling; this occurs partly through increased interactions between MUC16 and EGF-type (ErbB) receptors, which are enhanced by aberrant glycoforms of MUC16. Anti-MUC16 mAb AR9.6 blocks oncogenic signaling and reduces tumor growth in vivo.","method":"Isoform analysis, AKT/GSK3β phosphorylation Western blot, co-immunoprecipitation of MUC16 with ErbB receptors, mAb AR9.6 treatment in vitro and in mouse xenograft model","journal":"Molecular therapy","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP with functional blockade and in vivo validation, single lab","pmids":["33359791"],"is_preprint":false},{"year":2018,"finding":"MUC16 is cleaved at a specific location releasing the CA125 extracellular domain; the cleaved MUC16 subunits remain non-covalently associated with each other on the ovarian cancer cell surface, as demonstrated using antibodies specific to the retained carboxy-terminal juxtamembrane fragment.","method":"Generation and characterization of carboxy-terminus-specific monoclonal antibodies, immunoprecipitation, immunohistochemistry on ovarian tumor tissue","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 — novel antibody reagents with Co-IP confirming subunit association post-cleavage, single lab","pmids":["29708979"],"is_preprint":false},{"year":2019,"finding":"MUC16 facilitates cervical cancer cell proliferation and invasion via activation of JAK2/STAT3 phosphorylation, which promotes cyclooxygenase-2 (COX-2) expression; JAK2/STAT3 inhibition attenuates MUC16-driven COX-2 upregulation.","method":"shRNA knockdown and MUC16 overexpression, Western blot of JAK2/STAT3 phosphorylation, COX-2 expression, JAK2/STAT3 pharmacological inhibition, proliferation and invasion assays","journal":"Genes & genomics","confidence":"Medium","confidence_rationale":"Tier 2 — gain and loss of function with pathway inhibitor rescue, single lab","pmids":["31736008"],"is_preprint":false},{"year":2021,"finding":"MUC16 promotes fibrotic processes in idiopathic pulmonary fibrosis by collaborating with the TGF-β1 canonical pathway; MUC16 forms a protein complex with phospho-SMAD3 at the cell membrane after TGF-β1 stimulation, and siRNA-mediated MUC16 knockdown inhibits TGF-β1-induced SMAD3 phosphorylation, Smad Binding Element activation, and myofibroblast transformation.","method":"siRNA knockdown, immunoprecipitation of MUC16/p-SMAD3 complex, confocal immunofluorescence, SMAD3 phosphorylation assay, Smad Binding Element reporter, fibroblast proliferation and mesenchymal transformation assays","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP and colocalization with functional siRNA rescue, single lab","pmids":["34204432"],"is_preprint":false},{"year":2020,"finding":"MUC16 C-terminal domain (MUC16c) binds aldolase C (ALDOC), identified by mass spectrometry; MUC16c binding to ALDOC promotes ALDOC protein stability, disrupts ALDOC's ability to sense glucose deficiency, and activates the AMPK pathway to increase gallbladder carcinoma cell proliferation and glycolysis.","method":"Mass spectrometry interactome, co-immunoprecipitation, ALDOC knockdown rescue, glucose uptake and glycolysis assays, AMPK pathway Western blot, IHC","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 — MS identification plus Co-IP and functional rescue, single lab","pmids":["32502493"],"is_preprint":false},{"year":2022,"finding":"MUC16 promotes liver metastasis of pancreatic cancer by regulating Neuropilin-2 (NRP2) via JAK2/STAT1 signaling; NRP2 knockdown in MUC16-overexpressed cells decreases cell adhesion and migration; MUC16 also promotes endothelial/p-selectin binding and liver colonization in ex vivo and in vivo mouse models.","method":"MUC16 knockdown and MUC16-Cter ectopic overexpression, RNA-sequencing, JAK2/STAT1 pathway analysis, NRP2 knockdown rescue, ex vivo liver colonization, in vivo liver metastasis mouse model","journal":"Molecular cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — RNA-seq pathway identification with genetic rescue and in vivo validation, single lab","pmids":["35533267"],"is_preprint":false},{"year":2022,"finding":"Deletion of Muc16 in KRAS-driven and KRAS/p53 pancreatic cancer mouse models (KPCM, KCM) significantly decreases tumor progression and metastasis, and prolongs survival; MUC16 loss downregulates cytoskeletal proteins Actg2, Myh11, and Pdlim3, and knockdown of these genes reduces metastatic potential.","method":"Genetically engineered mouse models (KPC/KPCM), survival analysis, RNA-seq, syngeneic cell metastasis assays, organoid growth assay, cytoskeletal gene knockdown","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — in vivo genetic mouse model with mechanistic RNA-seq and validation of downstream effectors","pmids":["36271032"],"is_preprint":false},{"year":2022,"finding":"Truncated O-glycan (Tn and STn antigen)-bearing MUC16 interacts with α4β1 integrin complexes on pancreatic cancer cells; this interaction activates integrin-linked kinase and focal adhesion kinase (ILK/FAK) signaling and promotes tumor cell migration; CRISPR/Cas9 deletion of MUC16 or anti-MUC16 antibody reduces migration.","method":"CRISPR/Cas9 MUC16 deletion, co-immunoprecipitation of MUC16 with α4β1 integrins, FAK/ILK phosphorylation Western blot, migration assays, anti-MUC16 antibody blockade","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 — CRISPR KO plus Co-IP and functional blockade, single lab","pmids":["35628269"],"is_preprint":false},{"year":2023,"finding":"MUC16 promotes triple-negative breast cancer lung metastasis through a MUC16/HuR/cMyc axis; MUC16-Cter activates HuR (ELAVL1), which post-transcriptionally upregulates cMyc; RNA immunoprecipitation confirmed cMyc as a HuR target; MUC16 depletion or HuR pharmacological inhibition reduces cMyc expression and TNBC cell migration.","method":"MUC16 shRNA knockdown and MUC16-Cter overexpression, RNA immunoprecipitation (RIP) for HuR-cMyc association, microarray, tail vein in vivo metastasis model, HuR inhibitor (MS-444, CMLD-2) treatment","journal":"Breast cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — RIP mechanistic assay plus in vivo metastasis model and pharmacological rescue, single lab","pmids":["36918912"],"is_preprint":false},{"year":2022,"finding":"MUC16/CA125 shed from ovarian cancer tumors inhibits NK cell activation and cytotoxicity at the transcriptional level, suppressing expression of genes involved in NK cell activation and cytotoxicity pathways, as shown by RNA-sequencing of NK cells treated with ascites or CA125-enriched protein fractions.","method":"Fcγ receptor-mediated NK activation assay, RNA sequencing of NK cells from patient ascites and in vitro CA125-treated NK cells, CA125-enriched protein fraction preparation","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 — transcriptomic mechanism with CA125-specific fraction and matched patient samples, single lab","pmids":["35396222"],"is_preprint":false},{"year":2023,"finding":"MUC16 acts on neutrophils via Siglec-9 receptor to induce an inflammatory and immunosuppressive phenotype; MUC16 protein stimulation of neutrophils upregulates TNFA signaling, IL6-related pathways, and immunosuppression factors (PD-L1, IDO1, HHLA2), and supernatant from MUC16-stimulated neutrophils decreases NK cytotoxicity in vitro.","method":"MUC16 protein stimulation of neutrophils, flow cytometry, qPCR, RNA-sequencing, NK cytotoxicity assay with conditioned supernatant, Siglec-9 expression analysis on neutrophils","journal":"Journal of ovarian research","confidence":"Medium","confidence_rationale":"Tier 2 — direct protein stimulation with receptor identification and functional NK assay, single lab","pmids":["37644468"],"is_preprint":false},{"year":2009,"finding":"Targeted disruption of the Muc16 gene in mice (knockout) shows that Muc16 is dispensable for normal development, fertility, and organ histology; Muc16 null mice are viable and normal, with downregulation of Muc1 transcripts in the uterus.","method":"Gene targeting (deletion of exon 3 with lacZ reporter), histological analysis, RT-PCR for mucin gene expression, fertility assessment","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 — clean in vivo knockout mouse model with comprehensive phenotypic analysis","pmids":["19262696"],"is_preprint":false},{"year":2014,"finding":"Loss of Muc16 in knockout mice activates Stat3 signaling and upregulates IL-6 in the conjunctiva, affects JunB signaling, causes basal-like cell distribution in suprabasal corneal epithelium with increased proliferation, accelerates corneal epithelial wound healing, and increases myofibroblast appearance and macrophage invasion in stroma after epithelial repair.","method":"Muc16-null knockout mice, immunohistochemistry for phospho-Stat3/AP-1/IL-6/TNFα, BrdU proliferation labeling, corneal wound healing measurement, keratocyte phenotype assessment","journal":"Investigative ophthalmology & visual science","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo KO with multiple mechanistic readouts, single lab","pmids":["24812549"],"is_preprint":false}],"current_model":"MUC16 is a giant transmembrane mucin that functions as a physical barrier on epithelial surfaces (via ERM-mediated actin cytoskeleton linkage), mediates cell-cell adhesion through direct binding to mesothelin (via a defined 64-aa domain requiring tyrosine 318) and galectin-1 (via O-linked β-galactose chains), undergoes proteolytic shedding (by MT1-MMP) that releases the CA125 ectodomain while a retained C-terminal fragment signals oncogenically through JAK2/STAT3, Wnt/β-catenin, ErbB receptors (glycosylation-dependent), mTOR/c-Myc, and integrin/FAK pathways to drive cancer cell proliferation, invasion, and metabolic reprogramming, and suppresses innate immune responses (NK cells and macrophages/neutrophils) via Siglec-9 engagement and transcriptional suppression of cytotoxicity genes."},"narrative":{"teleology":[{"year":2003,"claim":"Identifying MUC16's binding partners established it as a cell-adhesion mediator rather than merely a passive mucin barrier, revealing that CA125/MUC16 directly binds mesothelin to mediate ovarian cancer–mesothelial cell adhesion and binds galectin-1 through O-linked β-galactose chains.","evidence":"Expression cloning, Co-IP, flow cytometry adhesion assays with antibody blockade (mesothelin); MS identification with glycan-specific binding assays (galectin-1)","pmids":["14676194","12615972"],"confidence":"High","gaps":["Cleavage site releasing CA125 ectodomain not yet mapped","Structural basis of MUC16–mesothelin interaction not resolved","In vivo significance of galectin-1 interaction not tested"]},{"year":2007,"claim":"MUC16 was shown to function as a dominant anti-adhesive barrier on uterine and corneal epithelial surfaces, resolving how the mucin glycocalyx contributes to epithelial protection at a molecular level.","evidence":"siRNA knockdown in uterine epithelial cells increasing trophoblast adhesion; RNAi in corneal epithelial cells reducing barrier function via dye exclusion and bacterial binding assays; peptide pull-down identifying ERM protein interaction with cytoplasmic tail","pmids":["17942799","17898272"],"confidence":"High","gaps":["Mechanism of MUC16 removal from uterodomes during implantation window unknown","Whether ERM linkage is required for barrier function not directly tested"]},{"year":2008,"claim":"Mapping the minimal mesothelin-binding region to 64 amino acids (residues 296–359) with critical dependence on tyrosine 318 provided the first molecular resolution of the MUC16–mesothelin adhesion interface.","evidence":"Truncation mutagenesis and alanine scanning with quantitative ELISA, Western blot overlay, and flow cytometry on cancer cells","pmids":["19075018"],"confidence":"High","gaps":["Reciprocal mapping of the binding site on MUC16 not performed","Crystal structure of the complex not determined"]},{"year":2009,"claim":"Muc16 knockout mice revealed that MUC16 is dispensable for normal development and fertility, redirecting attention to its context-dependent roles in disease rather than essential housekeeping function.","evidence":"Gene targeting (exon 3 deletion with lacZ reporter) in mice, comprehensive histological and fertility analysis","pmids":["19262696"],"confidence":"High","gaps":["Pathogen challenge and wound-healing phenotypes not assessed in initial characterization","Compensatory mucin upregulation beyond Muc1 not explored"]},{"year":2014,"claim":"Two advances established MUC16's dual role as a barrier determinant and a substrate for proteolytic shedding: comprehensive knockdown studies confirmed MUC16 as the dominant epithelial barrier mucin (over MUC1), while identification of MT1-MMP as the sheddase revealed how CA125 is released and how shedding switches cells from adhesive to invasive behavior.","evidence":"Stable shRNA knockdown with multiple barrier readouts (dye, bacteria, transepithelial resistance, tight junctions); MT1-MMP overexpression with catalytic-dead mutant control, peritoneal explant adhesion, and invasion assays","pmids":["24968021","25205731"],"confidence":"High","gaps":["Exact cleavage site within MUC16 not identified","Whether other MMPs contribute to shedding in vivo not resolved","Fate of the retained C-terminal fragment after shedding not characterized"]},{"year":2014,"claim":"Muc16-null mice showed activated Stat3/IL-6 signaling, increased corneal epithelial proliferation, and accelerated wound healing, establishing that MUC16 restrains inflammatory signaling in the ocular surface.","evidence":"Muc16-knockout mice with immunohistochemistry for phospho-STAT3, IL-6, BrdU proliferation, and corneal wound healing measurement","pmids":["24812549"],"confidence":"Medium","gaps":["Whether STAT3 activation is cell-autonomous or secondary to barrier loss not distinguished","Findings limited to ocular surface; relevance to other epithelia untested"]},{"year":2015,"claim":"Connecting MUC16 to metabolic reprogramming via the mTOR/c-Myc axis in pancreatic cancer established that MUC16 is not merely an adhesion molecule but also controls glycolysis and nucleotide metabolism.","evidence":"shRNA knockdown with c-MYC overexpression rescue, glucose uptake and lactate secretion assays, LC-MS/MS metabolomics in pancreatic cancer cells","pmids":["26046375"],"confidence":"Medium","gaps":["Direct molecular link between MUC16 and mTOR activation not identified","Metabolic reprogramming not validated in vivo"]},{"year":2016,"claim":"Discovery that the MUC16 C-terminal fragment (MUC16C) binds β-catenin and facilitates its nuclear translocation revealed a direct mechanism by which retained MUC16 activates Wnt signaling after ectodomain shedding.","evidence":"Co-IP with domain mapping, nuclear fractionation, Wnt reporter assays, MUC16 knockdown, and xenograft tumor model","pmids":["27167110"],"confidence":"Medium","gaps":["Whether MUC16C–β-catenin interaction requires prior shedding not tested","Structural basis of the interaction not resolved"]},{"year":2017,"claim":"Multiple studies converged to define MUC16 as a signaling platform whose activity depends on specific glycosylation: MGAT5-dependent N-glycosylation was required for galectin-3/growth-factor-receptor engagement on lipid rafts, the C-terminal domain activated JAK2/STAT3 to drive drug resistance, and oncogenic KRAS was shown to transcriptionally upregulate MUC16 via ERK/c-Myc, establishing a feed-forward loop.","evidence":"MGAT5 loss-of-function with glycan-specific antibodies, lipid raft fractionation, and xenograft model; JAK2/STAT3 inhibitor rescue with cisplatin/gemcitabine resistance assays; c-Myc ChIP on MUC16 promoter with ERK inhibitor epistasis","pmids":["28617578","28196872","28108627"],"confidence":"Medium","gaps":["Identity of specific growth factor receptors engaged via galectin-3 lattice incompletely defined","Whether KRAS–MUC16 feed-forward loop operates in non-pancreatic cancers unknown","Relative contributions of N- versus O-glycosylation to signaling not systematically compared"]},{"year":2019,"claim":"Functional immune assays demonstrated that MUC16 suppresses NK cell and macrophage cytotoxicity against ovarian cancer, directly linking ectodomain expression to immune evasion and in vivo survival advantage.","evidence":"NK and macrophage cytolysis assays, doublet formation assays, in vivo mouse survival with MUC16-knockdown tumors showing >2-fold increased survival","pmids":["30626487"],"confidence":"High","gaps":["Receptor on NK cells mediating MUC16 recognition not identified in this study","Contribution of shed versus membrane-bound MUC16 to immune suppression not separated"]},{"year":2020,"claim":"MUC16 was connected to additional signaling partners: aberrant glycoforms enhance MUC16–ErbB receptor interaction to activate AKT/GSK3β in pancreatic cancer, while the C-terminal domain binds aldolase C to stabilize ALDOC protein and activate AMPK-driven glycolysis in gallbladder carcinoma.","evidence":"Co-IP of MUC16 with ErbB receptors and anti-MUC16 mAb blockade in xenografts; MS-identified MUC16c–ALDOC interaction with ALDOC knockdown rescue and glycolysis assays","pmids":["33359791","32502493"],"confidence":"Medium","gaps":["Which specific ErbB family members are primary MUC16 partners not resolved","ALDOC interaction validated only in gallbladder cancer context","Whether glycoform-dependent signaling differences are tissue-specific not established"]},{"year":2022,"claim":"Genetic mouse models and integrin studies provided the strongest in vivo evidence for MUC16's pro-metastatic role: Muc16 deletion in KRAS-driven pancreatic cancer mice significantly reduced tumor progression and metastasis by downregulating cytoskeletal effectors, while truncated O-glycan-bearing MUC16 was shown to activate integrin/FAK signaling to promote migration, and shed CA125 was found to transcriptionally suppress NK cell cytotoxicity genes.","evidence":"KPCM/KCM genetically engineered mouse models with survival analysis and RNA-seq; CRISPR MUC16 KO with integrin Co-IP and FAK/ILK phosphorylation; RNA-seq of NK cells treated with CA125-enriched fractions from patient ascites","pmids":["36271032","35628269","35396222"],"confidence":"High","gaps":["Cytoskeletal effectors (Actg2, Myh11, Pdlim3) validated only by knockdown — direct transcriptional regulation not shown","Whether integrin/FAK and immune-evasion mechanisms operate simultaneously in vivo not tested","NK cell receptor(s) engaged by shed CA125 not identified"]},{"year":2023,"claim":"Two studies extended MUC16's immune-suppressive and metastatic mechanisms: MUC16 engages Siglec-9 on neutrophils to induce PD-L1/IDO1-expressing immunosuppressive phenotypes, and the MUC16-Cter/HuR/c-Myc post-transcriptional axis drives triple-negative breast cancer lung metastasis.","evidence":"MUC16 protein stimulation of neutrophils with Siglec-9 analysis, RNA-seq, and NK functional assay; MUC16 shRNA/OE with RNA immunoprecipitation for HuR–cMyc, tail-vein metastasis model, and HuR inhibitor treatment","pmids":["37644468","36918912"],"confidence":"Medium","gaps":["Direct Siglec-9 blockade/knockout not performed to confirm requirement","HuR/c-Myc axis not validated in cancer types beyond TNBC","Whether Siglec-9 engagement requires specific glycoforms of MUC16 not determined"]},{"year":null,"claim":"Key unresolved questions include the precise MUC16 cleavage site for MT1-MMP, the structural basis of MUC16–mesothelin and MUC16–β-catenin interactions, how membrane-bound versus shed MUC16 differentially contribute to immune suppression versus oncogenic signaling in vivo, and whether the diverse downstream pathways represent tissue-specific or universal MUC16 functions.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural (crystallographic or cryo-EM) data exist for any MUC16 interaction","Relative contribution of each signaling pathway in genetically defined in vivo models not established","Cleavage site for MT1-MMP-mediated shedding not mapped at amino acid resolution"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098631","term_label":"cell adhesion mediator activity","supporting_discovery_ids":[0,2,11]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[1,20]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[12,23,24]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1,4,5,6,15]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[3,6,15,23]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7,8,9,14,16,19,21]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[12,23,24]},{"term_id":"R-HSA-1500931","term_label":"Cell-Cell communication","supporting_discovery_ids":[0,2,11]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[8,9,20,22]}],"complexes":[],"partners":["MSLN","LGALS1","CTNNB1","LGALS3","ITGB1","ELAVL1","ALDOC","EZR"],"other_free_text":[]},"mechanistic_narrative":"MUC16 is a giant transmembrane mucin that serves as a dominant physical barrier on epithelial surfaces and functions as a multifaceted signaling hub in cancer progression and immune evasion. Its extracellular domain mediates heterotypic cell adhesion through direct binding to mesothelin (via a 64-amino acid region requiring tyrosine 318) and galectin-1 (via O-linked β-galactose chains), while its cytoplasmic tail links to the actin cytoskeleton through ERM proteins to maintain epithelial barrier integrity, tight junctions, and resistance to bacterial adherence [PMID:14676194, PMID:12615972, PMID:17898272, PMID:24968021]. Proteolytic shedding by MT1-MMP releases the CA125 ectodomain, while a retained C-terminal fragment drives oncogenic signaling through JAK2/STAT3, Wnt/β-catenin, mTOR/c-Myc, and integrin/FAK pathways, promoting proliferation, glycolytic metabolic reprogramming, invasion, and metastasis in multiple cancer types [PMID:25205731, PMID:27167110, PMID:28196872, PMID:26046375, PMID:35628269, PMID:36271032]. Shed MUC16 suppresses innate immunity by transcriptionally repressing NK cell cytotoxicity genes and engaging Siglec-9 on neutrophils to induce immunosuppressive phenotypes including PD-L1 and IDO1 upregulation [PMID:30626487, PMID:35396222, PMID:37644468]."},"prefetch_data":{"uniprot":{"accession":"Q8WXI7","full_name":"Mucin-16","aliases":["Ovarian cancer-related tumor marker CA125","CA-125","Ovarian carcinoma antigen CA125"],"length_aa":14507,"mass_kda":1519.2,"function":"Thought to provide a protective, lubricating barrier against particles and infectious agents at mucosal surfaces","subcellular_location":"Cell membrane; Secreted, extracellular space","url":"https://www.uniprot.org/uniprotkb/Q8WXI7/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MUC16","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MUC16","total_profiled":1310},"omim":[{"mim_id":"606154","title":"MUCIN 16; MUC16","url":"https://www.omim.org/entry/606154"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"adipose tissue","ntpm":6.5},{"tissue":"cervix","ntpm":15.0},{"tissue":"fallopian tube","ntpm":5.5},{"tissue":"salivary gland","ntpm":5.7}],"url":"https://www.proteinatlas.org/search/MUC16"},"hgnc":{"alias_symbol":["CA125","FLJ14303","CA-125"],"prev_symbol":[]},"alphafold":{"accession":"Q8WXI7","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8WXI7","model_url":"","pae_url":"","plddt_mean":null},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MUC16","jax_strain_url":"https://www.jax.org/strain/search?query=MUC16"},"sequence":{"accession":"Q8WXI7","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8WXI7.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8WXI7/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8WXI7"}},"corpus_meta":[{"pmid":"14676194","id":"PMC_14676194","title":"Binding of ovarian cancer antigen CA125/MUC16 to mesothelin mediates cell adhesion.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14676194","citation_count":455,"is_preprint":false},{"pmid":"10228898","id":"PMC_10228898","title":"CA 125: the past and the future.","date":"1998","source":"The International journal of biological markers","url":"https://pubmed.ncbi.nlm.nih.gov/10228898","citation_count":318,"is_preprint":false},{"pmid":"3457709","id":"PMC_3457709","title":"Elevated serum concentrations of CA-125 in patients with advanced endometriosis.","date":"1986","source":"Fertility and sterility","url":"https://pubmed.ncbi.nlm.nih.gov/3457709","citation_count":313,"is_preprint":false},{"pmid":"16061277","id":"PMC_16061277","title":"Potential markers that complement expression of CA125 in epithelial ovarian cancer.","date":"2005","source":"Gynecologic oncology","url":"https://pubmed.ncbi.nlm.nih.gov/16061277","citation_count":272,"is_preprint":false},{"pmid":"11786729","id":"PMC_11786729","title":"The CA 125 gene: an extracellular superstructure dominated by repeat sequences.","date":"2001","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/11786729","citation_count":232,"is_preprint":false},{"pmid":"3465595","id":"PMC_3465595","title":"The use of CA-125 in the diagnosis and management of endometriosis.","date":"1986","source":"Fertility and sterility","url":"https://pubmed.ncbi.nlm.nih.gov/3465595","citation_count":186,"is_preprint":false},{"pmid":"11920644","id":"PMC_11920644","title":"Ovarian cancer antigen CA125 is encoded by the MUC16 mucin gene.","date":"2002","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/11920644","citation_count":186,"is_preprint":false},{"pmid":"17898272","id":"PMC_17898272","title":"Functions of MUC16 in corneal epithelial cells.","date":"2007","source":"Investigative ophthalmology & visual science","url":"https://pubmed.ncbi.nlm.nih.gov/17898272","citation_count":166,"is_preprint":false},{"pmid":"19075018","id":"PMC_19075018","title":"A binding domain on mesothelin for CA125/MUC16.","date":"2008","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19075018","citation_count":157,"is_preprint":false},{"pmid":"12615972","id":"PMC_12615972","title":"The cancer antigen CA125 represents a novel counter receptor for galectin-1.","date":"2003","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/12615972","citation_count":127,"is_preprint":false},{"pmid":"22066010","id":"PMC_22066010","title":"Pathobiological implications of MUC16 expression in pancreatic cancer.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22066010","citation_count":110,"is_preprint":false},{"pmid":"24968021","id":"PMC_24968021","title":"Comparison of the transmembrane mucins MUC1 and MUC16 in epithelial barrier function.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24968021","citation_count":105,"is_preprint":false},{"pmid":"22320398","id":"PMC_22320398","title":"Coexpression of MUC16 and mesothelin is related to the invasion process in pancreatic ductal adenocarcinoma.","date":"2012","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/22320398","citation_count":101,"is_preprint":false},{"pmid":"25002120","id":"PMC_25002120","title":"MUC16: molecular analysis and its functional implications in benign and malignant conditions.","date":"2014","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/25002120","citation_count":95,"is_preprint":false},{"pmid":"28196872","id":"PMC_28196872","title":"MUC16 Regulates TSPYL5 for Lung Cancer Cell Growth and Chemoresistance by Suppressing p53.","date":"2017","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/28196872","citation_count":87,"is_preprint":false},{"pmid":"17942799","id":"PMC_17942799","title":"MUC16 is lost from the uterodome (pinopode) surface of the receptive human endometrium: in vitro evidence that MUC16 is a barrier to trophoblast adherence.","date":"2007","source":"Biology of reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/17942799","citation_count":87,"is_preprint":false},{"pmid":"26527287","id":"PMC_26527287","title":"Understanding the Unique Attributes of MUC16 (CA125): Potential Implications in Targeted Therapy.","date":"2015","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/26527287","citation_count":78,"is_preprint":false},{"pmid":"18782111","id":"PMC_18782111","title":"Alterations of the ocular surface epithelial MUC16 and goblet cell MUC5AC in patients with atopic keratoconjunctivitis.","date":"2008","source":"Allergy","url":"https://pubmed.ncbi.nlm.nih.gov/18782111","citation_count":76,"is_preprint":false},{"pmid":"32845327","id":"PMC_32845327","title":"Association of MUC16 Mutation With Response to Immune Checkpoint Inhibitors in Solid Tumors.","date":"2020","source":"JAMA network open","url":"https://pubmed.ncbi.nlm.nih.gov/32845327","citation_count":75,"is_preprint":false},{"pmid":"8365529","id":"PMC_8365529","title":"What do we know about the origin of CA 125?","date":"1993","source":"European journal of obstetrics, gynecology, and reproductive biology","url":"https://pubmed.ncbi.nlm.nih.gov/8365529","citation_count":72,"is_preprint":false},{"pmid":"18555226","id":"PMC_18555226","title":"Neutrophil-to-lymphocyte ratio as an adjunct to CA-125 for the diagnosis of endometriosis.","date":"2008","source":"Fertility and sterility","url":"https://pubmed.ncbi.nlm.nih.gov/18555226","citation_count":67,"is_preprint":false},{"pmid":"15297171","id":"PMC_15297171","title":"CA125- and tumor-specific T-cell responses correlate with prolonged survival in oregovomab-treated recurrent ovarian cancer patients.","date":"2004","source":"Gynecologic oncology","url":"https://pubmed.ncbi.nlm.nih.gov/15297171","citation_count":66,"is_preprint":false},{"pmid":"22247801","id":"PMC_22247801","title":"Evaluation of ovarian cancer biomarkers HE4 and CA-125 in women presenting with a suspicious cystic ovarian mass.","date":"2011","source":"Journal of gynecologic oncology","url":"https://pubmed.ncbi.nlm.nih.gov/22247801","citation_count":65,"is_preprint":false},{"pmid":"9023557","id":"PMC_9023557","title":"Immunohistochemical phenotype of malignant mesothelioma: predictive value of CA125 and HBME-1 expression.","date":"1997","source":"Histopathology","url":"https://pubmed.ncbi.nlm.nih.gov/9023557","citation_count":64,"is_preprint":false},{"pmid":"26046375","id":"PMC_26046375","title":"MUC16-mediated activation of mTOR and c-Myc reprograms pancreatic cancer metabolism.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26046375","citation_count":61,"is_preprint":false},{"pmid":"1934471","id":"PMC_1934471","title":"CA-125 concentrations in malignant and nonmalignant disease.","date":"1991","source":"Clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/1934471","citation_count":61,"is_preprint":false},{"pmid":"10202133","id":"PMC_10202133","title":"CA 125: fundamental and clinical aspects.","date":"1999","source":"Seminars in cancer biology","url":"https://pubmed.ncbi.nlm.nih.gov/10202133","citation_count":59,"is_preprint":false},{"pmid":"22286058","id":"PMC_22286058","title":"Pathobiological implications of MUC16/CA125 expression in intrahepatic cholangiocarcinoma-mass forming type.","date":"2012","source":"Pathobiology : journal of immunopathology, molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/22286058","citation_count":58,"is_preprint":false},{"pmid":"27483328","id":"PMC_27483328","title":"Functional Consequences of Differential O-glycosylation of MUC1, MUC4, and MUC16 (Downstream Effects on Signaling).","date":"2016","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/27483328","citation_count":56,"is_preprint":false},{"pmid":"18637025","id":"PMC_18637025","title":"MUC16 expression during embryogenesis, in adult tissues, and ovarian cancer in the mouse.","date":"2008","source":"Differentiation; research in biological diversity","url":"https://pubmed.ncbi.nlm.nih.gov/18637025","citation_count":54,"is_preprint":false},{"pmid":"12144685","id":"PMC_12144685","title":"Tissue and serum CA125 expression in endometrial cancer.","date":"2002","source":"International journal of gynecological cancer : official journal of the International Gynecological Cancer Society","url":"https://pubmed.ncbi.nlm.nih.gov/12144685","citation_count":53,"is_preprint":false},{"pmid":"33359791","id":"PMC_33359791","title":"Isoforms of MUC16 activate oncogenic signaling through EGF receptors to enhance the progression of pancreatic cancer.","date":"2020","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/33359791","citation_count":52,"is_preprint":false},{"pmid":"31472701","id":"PMC_31472701","title":"Molecularly imprinted polymer SPE sensor for analysis of CA-125 on serum.","date":"2019","source":"Analytica chimica acta","url":"https://pubmed.ncbi.nlm.nih.gov/31472701","citation_count":52,"is_preprint":false},{"pmid":"28108627","id":"PMC_28108627","title":"Oncogenic KRAS Targets MUC16/CA125 in Pancreatic Ductal Adenocarcinoma.","date":"2017","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/28108627","citation_count":48,"is_preprint":false},{"pmid":"9329618","id":"PMC_9329618","title":"Modulation of CA-125 release by inflammatory cytokines in human peritoneal mesothelial and ovarian cancer cells.","date":"1997","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/9329618","citation_count":48,"is_preprint":false},{"pmid":"12170412","id":"PMC_12170412","title":"Diagnostic laparoscopy, serum CA125, and peritoneal metastasis in gastric cancer.","date":"2002","source":"Endoscopy","url":"https://pubmed.ncbi.nlm.nih.gov/12170412","citation_count":47,"is_preprint":false},{"pmid":"19122828","id":"PMC_19122828","title":"MUC16 expression in Sjogren's syndrome, KCS, and control subjects.","date":"2008","source":"Molecular vision","url":"https://pubmed.ncbi.nlm.nih.gov/19122828","citation_count":42,"is_preprint":false},{"pmid":"28213228","id":"PMC_28213228","title":"Smartphone-based immunosensor for CA125 detection.","date":"2017","source":"Talanta","url":"https://pubmed.ncbi.nlm.nih.gov/28213228","citation_count":42,"is_preprint":false},{"pmid":"30017407","id":"PMC_30017407","title":"Comparison of CA125, HE4, and ROMA index for ovarian cancer diagnosis.","date":"2018","source":"Current problems in cancer","url":"https://pubmed.ncbi.nlm.nih.gov/30017407","citation_count":42,"is_preprint":false},{"pmid":"25205731","id":"PMC_25205731","title":"Membrane-type I matrix metalloproteinase-dependent ectodomain shedding of mucin16/ CA-125 on ovarian cancer cells modulates adhesion and invasion of peritoneal mesothelium.","date":"2014","source":"Biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25205731","citation_count":42,"is_preprint":false},{"pmid":"36552994","id":"PMC_36552994","title":"The Potential Role of MUC16 (CA125) Biomarker in Lung Cancer: A Magic Biomarker but with Adversity.","date":"2022","source":"Diagnostics (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/36552994","citation_count":41,"is_preprint":false},{"pmid":"24204560","id":"PMC_24204560","title":"MicroRNA-200c modulates the expression of MUC4 and MUC16 by directly targeting their coding sequences in human pancreatic cancer.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24204560","citation_count":41,"is_preprint":false},{"pmid":"24528057","id":"PMC_24528057","title":"CEA, AFP, CA125, CA153 and CA199 in malignant pleural effusions predict the cause.","date":"2014","source":"Asian Pacific journal of cancer prevention : APJCP","url":"https://pubmed.ncbi.nlm.nih.gov/24528057","citation_count":41,"is_preprint":false},{"pmid":"38361923","id":"PMC_38361923","title":"MUC1 and MUC16: critical for immune modulation in cancer therapeutics.","date":"2024","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/38361923","citation_count":38,"is_preprint":false},{"pmid":"27602187","id":"PMC_27602187","title":"Evaluation of HE4, CA-125, Risk of Ovarian Malignancy Algorithm (ROMA) and Risk of Malignancy Index (RMI) in the Preoperative Assessment of Patients with Adnexal Mass.","date":"2016","source":"Oman medical journal","url":"https://pubmed.ncbi.nlm.nih.gov/27602187","citation_count":38,"is_preprint":false},{"pmid":"31514860","id":"PMC_31514860","title":"A biosensor for determination of the circulating biomarker CA125/MUC16 by Surface Plasmon Resonance Imaging.","date":"2019","source":"Talanta","url":"https://pubmed.ncbi.nlm.nih.gov/31514860","citation_count":38,"is_preprint":false},{"pmid":"29241375","id":"PMC_29241375","title":"Peritoneal dissemination of ovarian cancer: role of MUC16-mesothelin interaction and implications for treatment.","date":"2017","source":"Expert review of anticancer therapy","url":"https://pubmed.ncbi.nlm.nih.gov/29241375","citation_count":36,"is_preprint":false},{"pmid":"9679738","id":"PMC_9679738","title":"Peritoneum and tissues of the female reproductive tract as physiological sources of CA-125.","date":"1998","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/9679738","citation_count":35,"is_preprint":false},{"pmid":"28617578","id":"PMC_28617578","title":"Antibodies Against Specific MUC16 Glycosylation Sites Inhibit Ovarian Cancer Growth.","date":"2017","source":"ACS chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/28617578","citation_count":34,"is_preprint":false},{"pmid":"31736008","id":"PMC_31736008","title":"MUC16 facilitates cervical cancer progression via JAK2/STAT3 phosphorylation-mediated cyclooxygenase-2 expression.","date":"2019","source":"Genes & genomics","url":"https://pubmed.ncbi.nlm.nih.gov/31736008","citation_count":33,"is_preprint":false},{"pmid":"34907079","id":"PMC_34907079","title":"ImmunoPET of Ovarian and Pancreatic Cancer with AR9.6, a Novel MUC16-Targeted Therapeutic Antibody.","date":"2022","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/34907079","citation_count":33,"is_preprint":false},{"pmid":"18342144","id":"PMC_18342144","title":"Regulation of MUC16 by inflammatory mediators in ocular surface epithelial cell lines.","date":"2007","source":"Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft","url":"https://pubmed.ncbi.nlm.nih.gov/18342144","citation_count":33,"is_preprint":false},{"pmid":"27167110","id":"PMC_27167110","title":"C-terminus of MUC16 activates Wnt signaling pathway through its interaction with β-catenin to promote tumorigenesis and metastasis.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27167110","citation_count":33,"is_preprint":false},{"pmid":"21525015","id":"PMC_21525015","title":"Tumour biomarkers in heart failure: is there a role for CA-125?","date":"2011","source":"European journal of heart failure","url":"https://pubmed.ncbi.nlm.nih.gov/21525015","citation_count":32,"is_preprint":false},{"pmid":"22949880","id":"PMC_22949880","title":"Deciphering the molecular nature of ovarian cancer biomarker CA125.","date":"2012","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/22949880","citation_count":32,"is_preprint":false},{"pmid":"19262696","id":"PMC_19262696","title":"CA125/MUC16 is dispensable for mouse development and reproduction.","date":"2009","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/19262696","citation_count":30,"is_preprint":false},{"pmid":"26390703","id":"PMC_26390703","title":"Diagnostic performances of CA125, HE4, and ROMA index in ovarian cancer.","date":"2015","source":"European journal of gynaecological oncology","url":"https://pubmed.ncbi.nlm.nih.gov/26390703","citation_count":30,"is_preprint":false},{"pmid":"11860541","id":"PMC_11860541","title":"Use of CA-125 and ultrasound in high-risk women.","date":"2002","source":"International journal of gynecological cancer : official journal of the International Gynecological Cancer Society","url":"https://pubmed.ncbi.nlm.nih.gov/11860541","citation_count":29,"is_preprint":false},{"pmid":"29609043","id":"PMC_29609043","title":"Evaluation of HE4 and TTR for diagnosis of ovarian cancer: Comparison with CA-125.","date":"2018","source":"Journal of gynecology obstetrics and human reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/29609043","citation_count":28,"is_preprint":false},{"pmid":"30626487","id":"PMC_30626487","title":"MUC16 suppresses human and murine innate immune responses.","date":"2019","source":"Gynecologic oncology","url":"https://pubmed.ncbi.nlm.nih.gov/30626487","citation_count":27,"is_preprint":false},{"pmid":"32502493","id":"PMC_32502493","title":"MUC16 C-terminal binding with ALDOC disrupts the ability of ALDOC to sense glucose and promotes gallbladder carcinoma growth.","date":"2020","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/32502493","citation_count":27,"is_preprint":false},{"pmid":"24812549","id":"PMC_24812549","title":"Effects of the loss of conjunctival Muc16 on corneal epithelium and stroma in mice.","date":"2014","source":"Investigative ophthalmology & visual science","url":"https://pubmed.ncbi.nlm.nih.gov/24812549","citation_count":27,"is_preprint":false},{"pmid":"35533267","id":"PMC_35533267","title":"MUC16 Promotes Liver Metastasis of Pancreatic Ductal Adenocarcinoma by Upregulating NRP2-Associated Cell Adhesion.","date":"2022","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/35533267","citation_count":26,"is_preprint":false},{"pmid":"28848147","id":"PMC_28848147","title":"Evaluation of Transvaginal Ultrasound plus CA-125 Measurement and Prophylactic Salpingo-Oophorectomy in Women at Different Risk Levels of Ovarian Cancer: The Modena Study Group Cohort Study.","date":"2017","source":"Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/28848147","citation_count":26,"is_preprint":false},{"pmid":"37644468","id":"PMC_37644468","title":"MUC16 stimulates neutrophils to an inflammatory and immunosuppressive phenotype in ovarian cancer.","date":"2023","source":"Journal of ovarian research","url":"https://pubmed.ncbi.nlm.nih.gov/37644468","citation_count":25,"is_preprint":false},{"pmid":"28576630","id":"PMC_28576630","title":"CA-125 in Disease Progression and Treatment of Lymphangioleiomyomatosis.","date":"2017","source":"Chest","url":"https://pubmed.ncbi.nlm.nih.gov/28576630","citation_count":24,"is_preprint":false},{"pmid":"28761326","id":"PMC_28761326","title":"Concentrations of MUC16 and MUC5AC using three tear collection methods.","date":"2017","source":"Molecular vision","url":"https://pubmed.ncbi.nlm.nih.gov/28761326","citation_count":24,"is_preprint":false},{"pmid":"36918912","id":"PMC_36918912","title":"MUC16 promotes triple-negative breast cancer lung metastasis by modulating RNA-binding protein ELAVL1/HUR.","date":"2023","source":"Breast cancer research : BCR","url":"https://pubmed.ncbi.nlm.nih.gov/36918912","citation_count":23,"is_preprint":false},{"pmid":"36813666","id":"PMC_36813666","title":"In silico designed mRNA vaccines targeting CA-125 neoantigen in breast and ovarian cancer.","date":"2023","source":"Vaccine","url":"https://pubmed.ncbi.nlm.nih.gov/36813666","citation_count":23,"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":"29708979","id":"PMC_29708979","title":"Development and characterization of carboxy-terminus specific monoclonal antibodies for understanding MUC16 cleavage in human ovarian cancer.","date":"2018","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/29708979","citation_count":23,"is_preprint":false},{"pmid":"10228901","id":"PMC_10228901","title":"CA 125 production and release by ovarian cancer cells in vitro.","date":"1998","source":"The International journal of biological markers","url":"https://pubmed.ncbi.nlm.nih.gov/10228901","citation_count":21,"is_preprint":false},{"pmid":"1832911","id":"PMC_1832911","title":"CA-125 in ovarian cancer: relation between half-life, doubling time and survival.","date":"1991","source":"European journal of cancer (Oxford, England : 1990)","url":"https://pubmed.ncbi.nlm.nih.gov/1832911","citation_count":21,"is_preprint":false},{"pmid":"26165164","id":"PMC_26165164","title":"Comparison of ovarian cancer markers in endometriosis favours HE4 over CA125.","date":"2015","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/26165164","citation_count":21,"is_preprint":false},{"pmid":"36271032","id":"PMC_36271032","title":"Muc16 depletion diminishes KRAS-induced tumorigenesis and metastasis by altering tumor microenvironment factors in pancreatic ductal adenocarcinoma.","date":"2022","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/36271032","citation_count":20,"is_preprint":false},{"pmid":"34204432","id":"PMC_34204432","title":"MUC16 Is Overexpressed in Idiopathic Pulmonary Fibrosis and Induces Fibrotic Responses Mediated by Transforming Growth Factor-β1 Canonical Pathway.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34204432","citation_count":20,"is_preprint":false},{"pmid":"28352780","id":"PMC_28352780","title":"The diagnosis and pathological value of combined detection of HE4 and CA125 for patients with ovarian cancer.","date":"2016","source":"Open medicine (Warsaw, Poland)","url":"https://pubmed.ncbi.nlm.nih.gov/28352780","citation_count":20,"is_preprint":false},{"pmid":"15788735","id":"PMC_15788735","title":"Introducing the MUC16 gene: implications for prevention and early detection in epithelial ovarian cancer.","date":"2005","source":"Biological research for nursing","url":"https://pubmed.ncbi.nlm.nih.gov/15788735","citation_count":19,"is_preprint":false},{"pmid":"24323399","id":"PMC_24323399","title":"HE4 combined with CA125: favorable screening tool for ovarian cancer.","date":"2013","source":"Medical oncology (Northwood, London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/24323399","citation_count":19,"is_preprint":false},{"pmid":"38758220","id":"PMC_38758220","title":"MUC16: clinical targets with great potential.","date":"2024","source":"Clinical and experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38758220","citation_count":19,"is_preprint":false},{"pmid":"34441373","id":"PMC_34441373","title":"Salvaging Detection of Early-Stage Ovarian Malignancies When CA125 Is Not Informative.","date":"2021","source":"Diagnostics (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/34441373","citation_count":19,"is_preprint":false},{"pmid":"20683153","id":"PMC_20683153","title":"Conflicting views on the molecular structure of the cancer antigen CA125/MUC16.","date":"2010","source":"Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/20683153","citation_count":18,"is_preprint":false},{"pmid":"26613889","id":"PMC_26613889","title":"CA-125, but not galectin-3, complements CA 19-9 for discriminating ductal adenocarcinoma versus non-malignant pancreatic diseases.","date":"2015","source":"Pancreatology : official journal of the International Association of Pancreatology (IAP) ... [et al.]","url":"https://pubmed.ncbi.nlm.nih.gov/26613889","citation_count":18,"is_preprint":false},{"pmid":"19589703","id":"PMC_19589703","title":"Premenarchal ovarian torsion and elevated CA-125.","date":"2009","source":"Journal of pediatric and adolescent gynecology","url":"https://pubmed.ncbi.nlm.nih.gov/19589703","citation_count":18,"is_preprint":false},{"pmid":"32977221","id":"PMC_32977221","title":"Concordance between CA-125 and RECIST progression in patients with germline BRCA-mutated platinum-sensitive relapsed ovarian cancer treated in the SOLO2 trial with olaparib as maintenance therapy after response to chemotherapy.","date":"2020","source":"European journal of cancer (Oxford, England : 1990)","url":"https://pubmed.ncbi.nlm.nih.gov/32977221","citation_count":17,"is_preprint":false},{"pmid":"31139280","id":"PMC_31139280","title":"The Biomarkers NT-proBNP and CA-125 are Elevated in Patients with Idiopathic Atrial Fibrillation.","date":"2018","source":"Journal of atrial fibrillation","url":"https://pubmed.ncbi.nlm.nih.gov/31139280","citation_count":17,"is_preprint":false},{"pmid":"35035759","id":"PMC_35035759","title":"Early diagonosis of ovarian cancer: serum HE4, CA125 and ROMA model.","date":"2021","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/35035759","citation_count":17,"is_preprint":false},{"pmid":"20566912","id":"PMC_20566912","title":"Expression of membrane-bound mucins in human nasal mucosa: different patterns for MUC4 and MUC16.","date":"2010","source":"Archives of otolaryngology--head & neck surgery","url":"https://pubmed.ncbi.nlm.nih.gov/20566912","citation_count":17,"is_preprint":false},{"pmid":"33832498","id":"PMC_33832498","title":"Significance of mesothelin and CA125 expression in endometrial carcinoma: a retrospective analysis.","date":"2021","source":"Diagnostic pathology","url":"https://pubmed.ncbi.nlm.nih.gov/33832498","citation_count":16,"is_preprint":false},{"pmid":"38050316","id":"PMC_38050316","title":"Diagnostic value of the combination of circulating serum miRNAs and CA125 in endometriosis.","date":"2023","source":"Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38050316","citation_count":16,"is_preprint":false},{"pmid":"35396222","id":"PMC_35396222","title":"Ovarian Cancer Ascites Inhibits Transcriptional Activation of NK Cells Partly through CA125.","date":"2022","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/35396222","citation_count":15,"is_preprint":false},{"pmid":"7741989","id":"PMC_7741989","title":"Markers supplementing CA 125 in ovarian cancer.","date":"1995","source":"Annals of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/7741989","citation_count":14,"is_preprint":false},{"pmid":"18644592","id":"PMC_18644592","title":"Differential expression of MUC16 in human oral mucosal epithelium and cultivated epithelial sheets.","date":"2008","source":"Experimental eye research","url":"https://pubmed.ncbi.nlm.nih.gov/18644592","citation_count":14,"is_preprint":false},{"pmid":"10228902","id":"PMC_10228902","title":"Regulation of CA 125 expression in cultured human carcinoma cells.","date":"1998","source":"The International journal of biological markers","url":"https://pubmed.ncbi.nlm.nih.gov/10228902","citation_count":13,"is_preprint":false},{"pmid":"37195923","id":"PMC_37195923","title":"NAV-001, a high-efficacy antibody-drug conjugate targeting mesothelin with improved delivery of a potent payload by counteracting MUC16/CA125 inhibitory effects.","date":"2023","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/37195923","citation_count":13,"is_preprint":false},{"pmid":"35884367","id":"PMC_35884367","title":"Urine CA125 and HE4 for the Triage of Symptomatic Women with Suspected Endometrial Cancer.","date":"2022","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/35884367","citation_count":13,"is_preprint":false},{"pmid":"8946867","id":"PMC_8946867","title":"Radiation-induced CA 125 production by mesothelial cells.","date":"1996","source":"Gynecologic oncology","url":"https://pubmed.ncbi.nlm.nih.gov/8946867","citation_count":13,"is_preprint":false},{"pmid":"10698036","id":"PMC_10698036","title":"CA-125 levels are related to the likelihood of pregnancy after in vitro fertilization and embryo transfer.","date":"2000","source":"American journal of reproductive immunology (New York, N.Y. : 1989)","url":"https://pubmed.ncbi.nlm.nih.gov/10698036","citation_count":13,"is_preprint":false},{"pmid":"35628269","id":"PMC_35628269","title":"Truncated O-Glycan-Bearing MUC16 Enhances Pancreatic Cancer Cells Aggressiveness via α4β1 Integrin Complexes and FAK Signaling.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35628269","citation_count":13,"is_preprint":false},{"pmid":"34150633","id":"PMC_34150633","title":"Knockdown of MUC16 (CA125) Enhances the Migration and Invasion of Hepatocellular Carcinoma Cells.","date":"2021","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34150633","citation_count":12,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":52153,"output_tokens":6686,"usd":0.128375},"stage2":{"model":"claude-opus-4-6","input_tokens":10365,"output_tokens":4116,"usd":0.232088},"total_usd":0.360463,"stage1_batch_id":"msgbatch_012ZiiCUBfB62NCTQd4Bk1Jo","stage2_batch_id":"msgbatch_01Xuqq9aUHoQouS7UVGZNStZ","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2003,\n      \"finding\": \"CA125/MUC16 binds to mesothelin, a GPI-anchored cell surface molecule, mediating heterotypic cell adhesion between ovarian cancer cells and mesothelial cells; anti-mesothelin antibody blocks this binding and cell attachment.\",\n      \"method\": \"Expression cloning, flow cytometry, immunoprecipitation, cell adhesion assay with antibody blockade\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP and functional adhesion assay with antibody blockade, replicated in multiple subsequent studies\",\n      \"pmids\": [\"14676194\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"MUC16 cytoplasmic tail binds the N-terminus of ERM (ezrin/radixin/moesin) actin-binding proteins, linking MUC16 to the actin cytoskeleton; MUC16 knockdown in corneal epithelial cells reduces barrier function (increased dye penetrance) and increases Staphylococcus aureus binding.\",\n      \"method\": \"Cytoplasmic tail peptide pull-down of ERM proteins, RNA interference knockdown, rose bengal dye exclusion assay, bacterial binding assay, scanning and immunoelectron microscopy\",\n      \"journal\": \"Investigative ophthalmology & visual science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct peptide pull-down plus functional RNAi phenotype with multiple orthogonal readouts\",\n      \"pmids\": [\"17898272\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"A 64-amino acid region (residues 296–359) at the N-terminal of cell-surface mesothelin is the minimum fragment sufficient for binding CA125/MUC16; tyrosine 318 substitution with alanine abolishes CA125 binding, and partial loss is caused by mutations at tryptophan 321 and glutamic acid 324.\",\n      \"method\": \"Truncated mutagenesis, alanine replacement, Western blot overlay assay, ELISA, flow cytometry on cancer cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro mutagenesis with quantitative binding assays and cell-based validation\",\n      \"pmids\": [\"19075018\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"CA125/MUC16 is a counter receptor for galectin-1; soluble and membrane-associated CA125 fragments bind galectin-1 specifically via beta-galactose-terminated O-linked oligosaccharide chains, with preference for galectin-1 over galectin-3, and CA125 expression enhances galectin-1 presentation on the cell surface.\",\n      \"method\": \"Mass spectrometry, immunological analysis, binding specificity assays with glycan inhibitors, FACS-based non-conventional secretion reconstitution assay\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — MS identification plus mechanistic glycan-dependent binding assays with functional reconstitution\",\n      \"pmids\": [\"12615972\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"MUC16 is lost from uterodome (pinopode) surfaces during the receptive phase of the uterine cycle (LH+6 to LH+8); siRNA knockdown of MUC16 (but not MUC1) in ECC-1 uterine epithelial cells increased trophoblast cell adhesion, demonstrating MUC16 acts as a barrier to trophoblast adherence.\",\n      \"method\": \"Immunofluorescence microscopy on uterine biopsies, siRNA knockdown, trophoblast adhesion assay\",\n      \"journal\": \"Biology of reproduction\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — specific siRNA with functional adhesion readout and parallel MUC1 control showing specificity\",\n      \"pmids\": [\"17942799\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MUC16 knockdown in epithelial cells decreases all barrier functions tested (dye penetrance, bacterial adherence and invasion, transepithelial resistance, tight junction formation, and apical surface size), while MUC1 knockdown increases barrier to dye and bacteria; demonstrating MUC16 provides the dominant epithelial barrier.\",\n      \"method\": \"Stable shRNA knockdown of MUC16 and MUC1 independently, dye penetrance assay, bacterial invasion assay, transepithelial resistance measurement, tight junction immunostaining\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with multiple orthogonal barrier readouts and internal control (MUC1 KD)\",\n      \"pmids\": [\"24968021\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MT1-MMP (MMP-14) mediates ectodomain shedding of MUC16/CA-125; overexpression of catalytically active MT1-MMP in OVCA433 cells causes loss of surface MUC16, while inactive E240A mutant does not; MUC16 shedding reduces adhesion to peritoneal tissue explants but enhances meso-mimetic invasion.\",\n      \"method\": \"MT1-MMP overexpression and catalytic mutant (E240A) experiments, surface MUC16 immunoreactivity, 3D meso-mimetic culture adhesion assay, ex vivo peritoneal explant adhesion assay, invasion assay\",\n      \"journal\": \"Biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — catalytic mutant controls plus multiple functional readouts in physiologically relevant model\",\n      \"pmids\": [\"25205731\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The C-terminal fragment of MUC16 (MUC16C) interacts with β-catenin (requiring β-catenin's trans-activation domain); this interaction facilitates cytosol-to-nucleus translocation of β-catenin, activating Wnt/β-catenin signaling to promote cell proliferation and migration, and tumorigenesis in nude mice.\",\n      \"method\": \"Co-immunoprecipitation with endogenous proteins using self-made MUC16 monoclonal antibody, domain-mapping, nuclear fractionation, reporter assays, MUC16 knockdown, xenograft tumor model\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP with domain mapping and in vivo validation, single lab\",\n      \"pmids\": [\"27167110\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"The C-terminal domain of MUC16 activates JAK2/STAT3/glucocorticoid receptor (GR) signaling to upregulate TSPYL5 in lung cancer cells; STAT3 inhibition reduces GR and TSPYL5; MUC16 overexpression induces cisplatin and gemcitabine resistance by downregulating p53.\",\n      \"method\": \"Stable shRNA knockdown and MUC16-Cter overexpression, transcriptome analysis, rescue experiments, STAT3 inhibitor treatment, Western blot of signaling proteins, in vivo tumor growth assay\",\n      \"journal\": \"Clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KD/OE with pathway rescue and in vivo validation, single lab\",\n      \"pmids\": [\"28196872\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"MUC16 knockdown in pancreatic cancer cells reduces mTOR activity and c-MYC expression; ectopic c-MYC restores the metabolic and physiological alterations caused by MUC16 knockdown, including reduced glucose uptake, lactate secretion, and motility. Metabolomics showed MUC16 promotes glycolytic and nucleotide metabolite pools.\",\n      \"method\": \"shRNA knockdown, c-MYC overexpression rescue, glucose uptake and lactate secretion assays, LC-MS/MS metabolomics, Western blot for mTOR activity\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — metabolomics plus genetic rescue, single lab\",\n      \"pmids\": [\"26046375\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Oncogenic KRAS upregulates MUC16 expression and CA125 shedding via the ERK/c-Myc axis in pancreatic cancer; c-Myc binds to the MUC16 promoter to transcriptionally activate its expression.\",\n      \"method\": \"KRAS manipulation in vitro and in vivo, ERK inhibitor treatment, c-Myc ChIP on MUC16 promoter, correlation analyses\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — promoter ChIP plus pathway epistasis, single lab\",\n      \"pmids\": [\"28108627\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"MUC16 and mesothelin are co-expressed and physically interact (co-immunoprecipitation) in pancreatic ductal adenocarcinoma; MUC16 shRNA knockdown and antibody blockade of MUC16-mesothelin binding inhibit invasion and migration of pancreatic cancer cells.\",\n      \"method\": \"Microarray, immunohistochemistry, co-immunoprecipitation from pancreatic cancer tissue, shRNA knockdown, antibody blockade, invasion and migration assays\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP from native tissue plus functional knockdown and antibody blockade with invasion readout\",\n      \"pmids\": [\"22320398\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MUC16 suppresses human and murine NK cell cytolysis and NK-tumor conjugate formation; MUC16-knockdown OVCAR-3 cells show increased susceptibility to NK and macrophage killing; mice bearing MUC16-knockdown tumors show >2-fold increased survival. MUC16 also suppresses ADCC by murine splenocytes.\",\n      \"method\": \"Target cell cytolysis assays, doublet formation assays, in vivo mouse survival model with MUC16-knockdown cells, in vitro NK/macrophage cytotoxicity assays\",\n      \"journal\": \"Gynecologic oncology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal immune cytotoxicity assays and in vivo model, replicates human and murine systems\",\n      \"pmids\": [\"30626487\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"MUC16 oncogenic signaling through its C-terminal ectodomain requires MGAT5-dependent N-glycosylation at two specific asparagine sites; oncogenic effects depend on Galectin-3 and growth factor receptors colocalized on lipid rafts; N-glycosylation site-directed antibodies block Galectin-3-mediated MUC16 interactions and inhibit ovarian cancer cell invasion and in vivo xenograft growth.\",\n      \"method\": \"MGAT5 loss-of-function, synthetic MUC16 glycopeptide-based monoclonal antibodies, galectin-3 expression knockdown, lipid raft fractionation, in vitro invasion assay, in vivo xenograft model\",\n      \"journal\": \"ACS chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — glycan-specific mutagenesis/ablation plus in vitro and in vivo functional validation with mechanistic antibodies\",\n      \"pmids\": [\"28617578\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"MUC16 enhances pancreatic cancer tumor malignancy through activation of AKT and GSK3β oncogenic signaling; this occurs partly through increased interactions between MUC16 and EGF-type (ErbB) receptors, which are enhanced by aberrant glycoforms of MUC16. Anti-MUC16 mAb AR9.6 blocks oncogenic signaling and reduces tumor growth in vivo.\",\n      \"method\": \"Isoform analysis, AKT/GSK3β phosphorylation Western blot, co-immunoprecipitation of MUC16 with ErbB receptors, mAb AR9.6 treatment in vitro and in mouse xenograft model\",\n      \"journal\": \"Molecular therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP with functional blockade and in vivo validation, single lab\",\n      \"pmids\": [\"33359791\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"MUC16 is cleaved at a specific location releasing the CA125 extracellular domain; the cleaved MUC16 subunits remain non-covalently associated with each other on the ovarian cancer cell surface, as demonstrated using antibodies specific to the retained carboxy-terminal juxtamembrane fragment.\",\n      \"method\": \"Generation and characterization of carboxy-terminus-specific monoclonal antibodies, immunoprecipitation, immunohistochemistry on ovarian tumor tissue\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — novel antibody reagents with Co-IP confirming subunit association post-cleavage, single lab\",\n      \"pmids\": [\"29708979\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MUC16 facilitates cervical cancer cell proliferation and invasion via activation of JAK2/STAT3 phosphorylation, which promotes cyclooxygenase-2 (COX-2) expression; JAK2/STAT3 inhibition attenuates MUC16-driven COX-2 upregulation.\",\n      \"method\": \"shRNA knockdown and MUC16 overexpression, Western blot of JAK2/STAT3 phosphorylation, COX-2 expression, JAK2/STAT3 pharmacological inhibition, proliferation and invasion assays\",\n      \"journal\": \"Genes & genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — gain and loss of function with pathway inhibitor rescue, single lab\",\n      \"pmids\": [\"31736008\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MUC16 promotes fibrotic processes in idiopathic pulmonary fibrosis by collaborating with the TGF-β1 canonical pathway; MUC16 forms a protein complex with phospho-SMAD3 at the cell membrane after TGF-β1 stimulation, and siRNA-mediated MUC16 knockdown inhibits TGF-β1-induced SMAD3 phosphorylation, Smad Binding Element activation, and myofibroblast transformation.\",\n      \"method\": \"siRNA knockdown, immunoprecipitation of MUC16/p-SMAD3 complex, confocal immunofluorescence, SMAD3 phosphorylation assay, Smad Binding Element reporter, fibroblast proliferation and mesenchymal transformation assays\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP and colocalization with functional siRNA rescue, single lab\",\n      \"pmids\": [\"34204432\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"MUC16 C-terminal domain (MUC16c) binds aldolase C (ALDOC), identified by mass spectrometry; MUC16c binding to ALDOC promotes ALDOC protein stability, disrupts ALDOC's ability to sense glucose deficiency, and activates the AMPK pathway to increase gallbladder carcinoma cell proliferation and glycolysis.\",\n      \"method\": \"Mass spectrometry interactome, co-immunoprecipitation, ALDOC knockdown rescue, glucose uptake and glycolysis assays, AMPK pathway Western blot, IHC\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — MS identification plus Co-IP and functional rescue, single lab\",\n      \"pmids\": [\"32502493\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MUC16 promotes liver metastasis of pancreatic cancer by regulating Neuropilin-2 (NRP2) via JAK2/STAT1 signaling; NRP2 knockdown in MUC16-overexpressed cells decreases cell adhesion and migration; MUC16 also promotes endothelial/p-selectin binding and liver colonization in ex vivo and in vivo mouse models.\",\n      \"method\": \"MUC16 knockdown and MUC16-Cter ectopic overexpression, RNA-sequencing, JAK2/STAT1 pathway analysis, NRP2 knockdown rescue, ex vivo liver colonization, in vivo liver metastasis mouse model\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — RNA-seq pathway identification with genetic rescue and in vivo validation, single lab\",\n      \"pmids\": [\"35533267\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Deletion of Muc16 in KRAS-driven and KRAS/p53 pancreatic cancer mouse models (KPCM, KCM) significantly decreases tumor progression and metastasis, and prolongs survival; MUC16 loss downregulates cytoskeletal proteins Actg2, Myh11, and Pdlim3, and knockdown of these genes reduces metastatic potential.\",\n      \"method\": \"Genetically engineered mouse models (KPC/KPCM), survival analysis, RNA-seq, syngeneic cell metastasis assays, organoid growth assay, cytoskeletal gene knockdown\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic mouse model with mechanistic RNA-seq and validation of downstream effectors\",\n      \"pmids\": [\"36271032\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Truncated O-glycan (Tn and STn antigen)-bearing MUC16 interacts with α4β1 integrin complexes on pancreatic cancer cells; this interaction activates integrin-linked kinase and focal adhesion kinase (ILK/FAK) signaling and promotes tumor cell migration; CRISPR/Cas9 deletion of MUC16 or anti-MUC16 antibody reduces migration.\",\n      \"method\": \"CRISPR/Cas9 MUC16 deletion, co-immunoprecipitation of MUC16 with α4β1 integrins, FAK/ILK phosphorylation Western blot, migration assays, anti-MUC16 antibody blockade\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — CRISPR KO plus Co-IP and functional blockade, single lab\",\n      \"pmids\": [\"35628269\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MUC16 promotes triple-negative breast cancer lung metastasis through a MUC16/HuR/cMyc axis; MUC16-Cter activates HuR (ELAVL1), which post-transcriptionally upregulates cMyc; RNA immunoprecipitation confirmed cMyc as a HuR target; MUC16 depletion or HuR pharmacological inhibition reduces cMyc expression and TNBC cell migration.\",\n      \"method\": \"MUC16 shRNA knockdown and MUC16-Cter overexpression, RNA immunoprecipitation (RIP) for HuR-cMyc association, microarray, tail vein in vivo metastasis model, HuR inhibitor (MS-444, CMLD-2) treatment\",\n      \"journal\": \"Breast cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — RIP mechanistic assay plus in vivo metastasis model and pharmacological rescue, single lab\",\n      \"pmids\": [\"36918912\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MUC16/CA125 shed from ovarian cancer tumors inhibits NK cell activation and cytotoxicity at the transcriptional level, suppressing expression of genes involved in NK cell activation and cytotoxicity pathways, as shown by RNA-sequencing of NK cells treated with ascites or CA125-enriched protein fractions.\",\n      \"method\": \"Fcγ receptor-mediated NK activation assay, RNA sequencing of NK cells from patient ascites and in vitro CA125-treated NK cells, CA125-enriched protein fraction preparation\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — transcriptomic mechanism with CA125-specific fraction and matched patient samples, single lab\",\n      \"pmids\": [\"35396222\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MUC16 acts on neutrophils via Siglec-9 receptor to induce an inflammatory and immunosuppressive phenotype; MUC16 protein stimulation of neutrophils upregulates TNFA signaling, IL6-related pathways, and immunosuppression factors (PD-L1, IDO1, HHLA2), and supernatant from MUC16-stimulated neutrophils decreases NK cytotoxicity in vitro.\",\n      \"method\": \"MUC16 protein stimulation of neutrophils, flow cytometry, qPCR, RNA-sequencing, NK cytotoxicity assay with conditioned supernatant, Siglec-9 expression analysis on neutrophils\",\n      \"journal\": \"Journal of ovarian research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct protein stimulation with receptor identification and functional NK assay, single lab\",\n      \"pmids\": [\"37644468\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Targeted disruption of the Muc16 gene in mice (knockout) shows that Muc16 is dispensable for normal development, fertility, and organ histology; Muc16 null mice are viable and normal, with downregulation of Muc1 transcripts in the uterus.\",\n      \"method\": \"Gene targeting (deletion of exon 3 with lacZ reporter), histological analysis, RT-PCR for mucin gene expression, fertility assessment\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean in vivo knockout mouse model with comprehensive phenotypic analysis\",\n      \"pmids\": [\"19262696\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Loss of Muc16 in knockout mice activates Stat3 signaling and upregulates IL-6 in the conjunctiva, affects JunB signaling, causes basal-like cell distribution in suprabasal corneal epithelium with increased proliferation, accelerates corneal epithelial wound healing, and increases myofibroblast appearance and macrophage invasion in stroma after epithelial repair.\",\n      \"method\": \"Muc16-null knockout mice, immunohistochemistry for phospho-Stat3/AP-1/IL-6/TNFα, BrdU proliferation labeling, corneal wound healing measurement, keratocyte phenotype assessment\",\n      \"journal\": \"Investigative ophthalmology & visual science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo KO with multiple mechanistic readouts, single lab\",\n      \"pmids\": [\"24812549\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MUC16 is a giant transmembrane mucin that functions as a physical barrier on epithelial surfaces (via ERM-mediated actin cytoskeleton linkage), mediates cell-cell adhesion through direct binding to mesothelin (via a defined 64-aa domain requiring tyrosine 318) and galectin-1 (via O-linked β-galactose chains), undergoes proteolytic shedding (by MT1-MMP) that releases the CA125 ectodomain while a retained C-terminal fragment signals oncogenically through JAK2/STAT3, Wnt/β-catenin, ErbB receptors (glycosylation-dependent), mTOR/c-Myc, and integrin/FAK pathways to drive cancer cell proliferation, invasion, and metabolic reprogramming, and suppresses innate immune responses (NK cells and macrophages/neutrophils) via Siglec-9 engagement and transcriptional suppression of cytotoxicity genes.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"MUC16 is a giant transmembrane mucin that serves as a dominant physical barrier on epithelial surfaces and functions as a multifaceted signaling hub in cancer progression and immune evasion. Its extracellular domain mediates heterotypic cell adhesion through direct binding to mesothelin (via a 64-amino acid region requiring tyrosine 318) and galectin-1 (via O-linked β-galactose chains), while its cytoplasmic tail links to the actin cytoskeleton through ERM proteins to maintain epithelial barrier integrity, tight junctions, and resistance to bacterial adherence [PMID:14676194, PMID:12615972, PMID:17898272, PMID:24968021]. Proteolytic shedding by MT1-MMP releases the CA125 ectodomain, while a retained C-terminal fragment drives oncogenic signaling through JAK2/STAT3, Wnt/β-catenin, mTOR/c-Myc, and integrin/FAK pathways, promoting proliferation, glycolytic metabolic reprogramming, invasion, and metastasis in multiple cancer types [PMID:25205731, PMID:27167110, PMID:28196872, PMID:26046375, PMID:35628269, PMID:36271032]. Shed MUC16 suppresses innate immunity by transcriptionally repressing NK cell cytotoxicity genes and engaging Siglec-9 on neutrophils to induce immunosuppressive phenotypes including PD-L1 and IDO1 upregulation [PMID:30626487, PMID:35396222, PMID:37644468].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Identifying MUC16's binding partners established it as a cell-adhesion mediator rather than merely a passive mucin barrier, revealing that CA125/MUC16 directly binds mesothelin to mediate ovarian cancer–mesothelial cell adhesion and binds galectin-1 through O-linked β-galactose chains.\",\n      \"evidence\": \"Expression cloning, Co-IP, flow cytometry adhesion assays with antibody blockade (mesothelin); MS identification with glycan-specific binding assays (galectin-1)\",\n      \"pmids\": [\"14676194\", \"12615972\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Cleavage site releasing CA125 ectodomain not yet mapped\",\n        \"Structural basis of MUC16–mesothelin interaction not resolved\",\n        \"In vivo significance of galectin-1 interaction not tested\"\n      ]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"MUC16 was shown to function as a dominant anti-adhesive barrier on uterine and corneal epithelial surfaces, resolving how the mucin glycocalyx contributes to epithelial protection at a molecular level.\",\n      \"evidence\": \"siRNA knockdown in uterine epithelial cells increasing trophoblast adhesion; RNAi in corneal epithelial cells reducing barrier function via dye exclusion and bacterial binding assays; peptide pull-down identifying ERM protein interaction with cytoplasmic tail\",\n      \"pmids\": [\"17942799\", \"17898272\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Mechanism of MUC16 removal from uterodomes during implantation window unknown\",\n        \"Whether ERM linkage is required for barrier function not directly tested\"\n      ]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Mapping the minimal mesothelin-binding region to 64 amino acids (residues 296–359) with critical dependence on tyrosine 318 provided the first molecular resolution of the MUC16–mesothelin adhesion interface.\",\n      \"evidence\": \"Truncation mutagenesis and alanine scanning with quantitative ELISA, Western blot overlay, and flow cytometry on cancer cells\",\n      \"pmids\": [\"19075018\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Reciprocal mapping of the binding site on MUC16 not performed\",\n        \"Crystal structure of the complex not determined\"\n      ]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Muc16 knockout mice revealed that MUC16 is dispensable for normal development and fertility, redirecting attention to its context-dependent roles in disease rather than essential housekeeping function.\",\n      \"evidence\": \"Gene targeting (exon 3 deletion with lacZ reporter) in mice, comprehensive histological and fertility analysis\",\n      \"pmids\": [\"19262696\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Pathogen challenge and wound-healing phenotypes not assessed in initial characterization\",\n        \"Compensatory mucin upregulation beyond Muc1 not explored\"\n      ]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Two advances established MUC16's dual role as a barrier determinant and a substrate for proteolytic shedding: comprehensive knockdown studies confirmed MUC16 as the dominant epithelial barrier mucin (over MUC1), while identification of MT1-MMP as the sheddase revealed how CA125 is released and how shedding switches cells from adhesive to invasive behavior.\",\n      \"evidence\": \"Stable shRNA knockdown with multiple barrier readouts (dye, bacteria, transepithelial resistance, tight junctions); MT1-MMP overexpression with catalytic-dead mutant control, peritoneal explant adhesion, and invasion assays\",\n      \"pmids\": [\"24968021\", \"25205731\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Exact cleavage site within MUC16 not identified\",\n        \"Whether other MMPs contribute to shedding in vivo not resolved\",\n        \"Fate of the retained C-terminal fragment after shedding not characterized\"\n      ]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Muc16-null mice showed activated Stat3/IL-6 signaling, increased corneal epithelial proliferation, and accelerated wound healing, establishing that MUC16 restrains inflammatory signaling in the ocular surface.\",\n      \"evidence\": \"Muc16-knockout mice with immunohistochemistry for phospho-STAT3, IL-6, BrdU proliferation, and corneal wound healing measurement\",\n      \"pmids\": [\"24812549\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether STAT3 activation is cell-autonomous or secondary to barrier loss not distinguished\",\n        \"Findings limited to ocular surface; relevance to other epithelia untested\"\n      ]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Connecting MUC16 to metabolic reprogramming via the mTOR/c-Myc axis in pancreatic cancer established that MUC16 is not merely an adhesion molecule but also controls glycolysis and nucleotide metabolism.\",\n      \"evidence\": \"shRNA knockdown with c-MYC overexpression rescue, glucose uptake and lactate secretion assays, LC-MS/MS metabolomics in pancreatic cancer cells\",\n      \"pmids\": [\"26046375\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct molecular link between MUC16 and mTOR activation not identified\",\n        \"Metabolic reprogramming not validated in vivo\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Discovery that the MUC16 C-terminal fragment (MUC16C) binds β-catenin and facilitates its nuclear translocation revealed a direct mechanism by which retained MUC16 activates Wnt signaling after ectodomain shedding.\",\n      \"evidence\": \"Co-IP with domain mapping, nuclear fractionation, Wnt reporter assays, MUC16 knockdown, and xenograft tumor model\",\n      \"pmids\": [\"27167110\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether MUC16C–β-catenin interaction requires prior shedding not tested\",\n        \"Structural basis of the interaction not resolved\"\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Multiple studies converged to define MUC16 as a signaling platform whose activity depends on specific glycosylation: MGAT5-dependent N-glycosylation was required for galectin-3/growth-factor-receptor engagement on lipid rafts, the C-terminal domain activated JAK2/STAT3 to drive drug resistance, and oncogenic KRAS was shown to transcriptionally upregulate MUC16 via ERK/c-Myc, establishing a feed-forward loop.\",\n      \"evidence\": \"MGAT5 loss-of-function with glycan-specific antibodies, lipid raft fractionation, and xenograft model; JAK2/STAT3 inhibitor rescue with cisplatin/gemcitabine resistance assays; c-Myc ChIP on MUC16 promoter with ERK inhibitor epistasis\",\n      \"pmids\": [\"28617578\", \"28196872\", \"28108627\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Identity of specific growth factor receptors engaged via galectin-3 lattice incompletely defined\",\n        \"Whether KRAS–MUC16 feed-forward loop operates in non-pancreatic cancers unknown\",\n        \"Relative contributions of N- versus O-glycosylation to signaling not systematically compared\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Functional immune assays demonstrated that MUC16 suppresses NK cell and macrophage cytotoxicity against ovarian cancer, directly linking ectodomain expression to immune evasion and in vivo survival advantage.\",\n      \"evidence\": \"NK and macrophage cytolysis assays, doublet formation assays, in vivo mouse survival with MUC16-knockdown tumors showing >2-fold increased survival\",\n      \"pmids\": [\"30626487\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Receptor on NK cells mediating MUC16 recognition not identified in this study\",\n        \"Contribution of shed versus membrane-bound MUC16 to immune suppression not separated\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"MUC16 was connected to additional signaling partners: aberrant glycoforms enhance MUC16–ErbB receptor interaction to activate AKT/GSK3β in pancreatic cancer, while the C-terminal domain binds aldolase C to stabilize ALDOC protein and activate AMPK-driven glycolysis in gallbladder carcinoma.\",\n      \"evidence\": \"Co-IP of MUC16 with ErbB receptors and anti-MUC16 mAb blockade in xenografts; MS-identified MUC16c–ALDOC interaction with ALDOC knockdown rescue and glycolysis assays\",\n      \"pmids\": [\"33359791\", \"32502493\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Which specific ErbB family members are primary MUC16 partners not resolved\",\n        \"ALDOC interaction validated only in gallbladder cancer context\",\n        \"Whether glycoform-dependent signaling differences are tissue-specific not established\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Genetic mouse models and integrin studies provided the strongest in vivo evidence for MUC16's pro-metastatic role: Muc16 deletion in KRAS-driven pancreatic cancer mice significantly reduced tumor progression and metastasis by downregulating cytoskeletal effectors, while truncated O-glycan-bearing MUC16 was shown to activate integrin/FAK signaling to promote migration, and shed CA125 was found to transcriptionally suppress NK cell cytotoxicity genes.\",\n      \"evidence\": \"KPCM/KCM genetically engineered mouse models with survival analysis and RNA-seq; CRISPR MUC16 KO with integrin Co-IP and FAK/ILK phosphorylation; RNA-seq of NK cells treated with CA125-enriched fractions from patient ascites\",\n      \"pmids\": [\"36271032\", \"35628269\", \"35396222\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Cytoskeletal effectors (Actg2, Myh11, Pdlim3) validated only by knockdown — direct transcriptional regulation not shown\",\n        \"Whether integrin/FAK and immune-evasion mechanisms operate simultaneously in vivo not tested\",\n        \"NK cell receptor(s) engaged by shed CA125 not identified\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Two studies extended MUC16's immune-suppressive and metastatic mechanisms: MUC16 engages Siglec-9 on neutrophils to induce PD-L1/IDO1-expressing immunosuppressive phenotypes, and the MUC16-Cter/HuR/c-Myc post-transcriptional axis drives triple-negative breast cancer lung metastasis.\",\n      \"evidence\": \"MUC16 protein stimulation of neutrophils with Siglec-9 analysis, RNA-seq, and NK functional assay; MUC16 shRNA/OE with RNA immunoprecipitation for HuR–cMyc, tail-vein metastasis model, and HuR inhibitor treatment\",\n      \"pmids\": [\"37644468\", \"36918912\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct Siglec-9 blockade/knockout not performed to confirm requirement\",\n        \"HuR/c-Myc axis not validated in cancer types beyond TNBC\",\n        \"Whether Siglec-9 engagement requires specific glycoforms of MUC16 not determined\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the precise MUC16 cleavage site for MT1-MMP, the structural basis of MUC16–mesothelin and MUC16–β-catenin interactions, how membrane-bound versus shed MUC16 differentially contribute to immune suppression versus oncogenic signaling in vivo, and whether the diverse downstream pathways represent tissue-specific or universal MUC16 functions.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural (crystallographic or cryo-EM) data exist for any MUC16 interaction\",\n        \"Relative contribution of each signaling pathway in genetically defined in vivo models not established\",\n        \"Cleavage site for MT1-MMP-mediated shedding not mapped at amino acid resolution\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098631\", \"supporting_discovery_ids\": [0, 2, 11]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [1, 20]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [12, 23, 24]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1, 4, 5, 6, 15]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [3, 6, 15, 23]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7, 8, 9, 14, 16, 19, 21]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [12, 23, 24]},\n      {\"term_id\": \"R-HSA-1500931\", \"supporting_discovery_ids\": [0, 2, 11]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [8, 9, 20, 22]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"MSLN\",\n      \"LGALS1\",\n      \"CTNNB1\",\n      \"LGALS3\",\n      \"ITGB1\",\n      \"ELAVL1\",\n      \"ALDOC\",\n      \"EZR\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}