{"gene":"MSLN","run_date":"2026-06-10T02:59:51","timeline":{"discoveries":[{"year":2000,"finding":"The MSLN gene spans 8 kb of human chromosome 16, is encoded by 15 exons with an 1884-bp open reading frame, and its promoter lacks a TATA box and SP1 sites. Transient transfection assays localized a minimal constitutive promoter to a 317-bp region and identified tissue-specific enhancer elements upstream of this minimal promoter that activate transcription in a position- and orientation-independent manner.","method":"Transient transfection analysis, promoter deletion constructs, genomic sequencing","journal":"Molecular cell biology research communications : MCBRC","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct promoter functional dissection with deletion constructs and transfection assays in a single study; no replication by independent lab","pmids":["10683314"],"is_preprint":false},{"year":2007,"finding":"An 18-bp upstream enhancer element called 'CanScript' drives cancer-specific MSLN overexpression. The CanScript contains two functional sites: an Sp1-like site and an MCAT element. Gel retardation and chromatin immunoprecipitation assays showed that transcription enhancer factor TEF-1 (TEAD1) binds the MCAT element both in vitro and in vivo, and TEF-1 knockdown reduced MSLN protein overexpression in cancer cells.","method":"Promoter deletion/mutation analysis, gel retardation (EMSA), chromatin immunoprecipitation (ChIP), TEF-1 siRNA knockdown, transient transfection","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (EMSA, ChIP, mutagenesis, knockdown) in one study establishing TEF-1/TEAD1 as direct transcriptional activator of MSLN via CanScript MCAT element","pmids":["17909009"],"is_preprint":false},{"year":2021,"finding":"MSLN knockout mice (Msln-/-) are protected from cholestatic liver fibrosis (Mdr2-deficient model), bleomycin-induced lung fibrosis, and UUO-induced kidney fibrosis, while Thy1-/- mice are more susceptible, indicating that a MSLN-Thy1 signaling complex is critical for tissue fibroblast activation. Targeting MSLN+ activated portal fibroblasts with anti-MSLN immunotoxins reduced collagen deposition in bile duct ligation-injured mice.","method":"Genetic knockout mouse models (Msln-/-, Thy1-/-), multiple fibrosis models (BDL, bleomycin, UUO), anti-MSLN immunotoxin treatment, collagen deposition assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function genetic models replicated across three distinct fibrosis models with defined cellular phenotype; functional rescue with immunotoxin","pmids":["34253615"],"is_preprint":false},{"year":2021,"finding":"Metformin treatment in ovarian cancer cells reduced MSLN expression, which in turn downregulated IL-6/STAT3 signaling activity and subsequently reduced VEGF and TGFβ1 expression, leading to inhibition of cell growth, migration, and stemness.","method":"Cell-based assays (proliferation, migration, apoptosis), molecular knockdown/overexpression, Western blot for pathway components","journal":"Cell transplantation","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement based on expression changes after metformin treatment; no direct reconstitution of MSLN–IL-6/STAT3 interaction","pmids":["34238029"],"is_preprint":false},{"year":2014,"finding":"Transient siRNA-mediated MSLN knockdown in MSLN-overexpressing mesothelioma cells (Mero-14) caused decreased proliferation, reduced invasive capacity, and reduced sphere formation; combined MSLN-siRNA with cisplatin triggered increased apoptosis and decreased proliferation compared to either agent alone.","method":"siRNA knockdown, proliferation assay, invasion assay, sphere formation assay, apoptosis assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — clean loss-of-function with defined cellular phenotypes (proliferation, invasion, sphere formation) across multiple readouts in a single study; replicated previous findings from other cell lines","pmids":["24465798"],"is_preprint":false},{"year":2014,"finding":"A common SNP (rs1057147) in the 3' UTR of MSLN creates a binding site for miR-611. Dual-luciferase reporter assays showed lower reporter activity with the G allele vs. A allele; miR-611 mimic reduced reporter activity only of the G-allele construct. Transfection of miR-611 into mesothelioma cells (Mero-14) significantly reduced endogenous MSLN protein levels, indicating post-transcriptional regulation of MSLN by miR-611 in a genotype-dependent manner.","method":"Dual-luciferase reporter assay with allele-specific constructs, miRNA mimic transfection, Western blot for endogenous MSLN protein","journal":"Journal of thoracic oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional reporter assay plus endogenous protein knockdown by miRNA in single lab; two orthogonal methods establish miR-611 as post-transcriptional regulator of MSLN","pmids":["25436799"],"is_preprint":false},{"year":2019,"finding":"MSLN-targeted thorium-227 conjugate (BAY 2287411) binds MSLN-expressing tumor cells and, upon delivery of alpha-particle radiation, induces DNA double-strand breaks, apoptotic markers, and oxidative stress, leading to reduced cellular viability and significant antitumor efficacy in vivo in multiple cancer xenograft models.","method":"In vitro binding assay, DNA damage markers (γH2AX), apoptosis assays, in vivo xenograft models (breast, colorectal, lung, ovarian, pancreatic), biodistribution studies","journal":"Clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple in vitro and in vivo models with defined mechanism of action (MSLN-targeted alpha-radiation causing DSBs); single lab but multiple orthogonal readouts","pmids":["31064781"],"is_preprint":false}],"current_model":"MSLN encodes mesothelin, a GPI-anchored membrane glycoprotein whose cancer-specific overexpression is driven by TEF-1/TEAD1 binding to an MCAT-containing enhancer element (CanScript) in its promoter; MSLN expression is also post-transcriptionally regulated by miR-611 in a SNP-dependent manner; functionally, MSLN forms a signaling complex with Thy1 that promotes tissue fibroblast activation across multiple organs, and loss of MSLN protects mice from fibrosis, while MSLN knockdown in cancer cells reduces proliferation, invasion, and stemness, partly through modulation of IL-6/STAT3 signaling."},"narrative":{"mechanistic_narrative":"MSLN encodes mesothelin, a cell-surface glycoprotein whose cancer-specific overexpression is driven transcriptionally by the enhancer factor TEF-1/TEAD1, which binds the MCAT element within an 18-bp upstream 'CanScript' enhancer; TEF-1 knockdown reduces mesothelin overexpression in cancer cells [PMID:17909009]. The MSLN promoter lacks TATA and SP1 boxes and depends on a minimal constitutive region plus position- and orientation-independent tissue-specific enhancers [PMID:10683314]. Mesothelin expression is additionally tuned post-transcriptionally: a common 3' UTR SNP (rs1057147) creates an allele-specific miR-611 binding site, and miR-611 reduces endogenous mesothelin protein in a genotype-dependent manner [PMID:25436799]. Functionally, mesothelin acts in a MSLN-Thy1 signaling axis that drives tissue fibroblast activation; Msln-knockout mice are protected from cholestatic liver, bleomycin lung, and UUO kidney fibrosis, and targeting MSLN+ portal fibroblasts reduces collagen deposition [PMID:34253615]. In cancer cells, loss of mesothelin diminishes proliferation, invasion, and sphere-forming stemness [PMID:24465798]. Its surface accessibility makes mesothelin a tractable therapeutic target, exemplified by a MSLN-directed thorium-227 alpha-particle conjugate that induces DNA double-strand breaks and antitumor efficacy across xenograft models [PMID:31064781].","teleology":[{"year":2000,"claim":"Defining the architecture of the MSLN promoter was needed to understand how the gene is transcriptionally controlled; this established a TATA-less minimal promoter governed by separable upstream tissue-specific enhancers.","evidence":"Promoter deletion constructs and transient transfection assays with genomic sequencing","pmids":["10683314"],"confidence":"Medium","gaps":["Identity of the trans-acting factors binding the enhancers not resolved","No link to cancer-specific expression at this stage","Single study without independent replication"]},{"year":2007,"claim":"It was unknown which factor drove cancer-selective MSLN overexpression; identification of TEF-1/TEAD1 binding the MCAT element of the CanScript enhancer established a direct transcriptional activator of MSLN in cancer.","evidence":"EMSA, ChIP, promoter mutagenesis, and TEF-1 siRNA knockdown in cancer cells","pmids":["17909009"],"confidence":"High","gaps":["Upstream signals controlling TEF-1 activity at this enhancer unknown","Role of the adjacent Sp1-like site not fully dissected","Whether the same mechanism operates in non-malignant tissue not addressed"]},{"year":2014,"claim":"Whether mesothelin had a cell-autonomous role in tumor biology was unclear; siRNA knockdown in mesothelioma cells established that mesothelin supports proliferation, invasion, and stemness and sensitizes cells to cisplatin.","evidence":"siRNA knockdown with proliferation, invasion, sphere formation, and apoptosis assays in Mero-14 cells","pmids":["24465798"],"confidence":"Medium","gaps":["Molecular effectors downstream of mesothelin not defined here","Transient knockdown only","In vivo relevance not tested"]},{"year":2014,"claim":"The basis for inter-individual variation in mesothelin levels was unknown; a 3' UTR SNP creating an allele-specific miR-611 site established post-transcriptional, genotype-dependent regulation of MSLN.","evidence":"Allele-specific dual-luciferase reporters, miR-611 mimic transfection, and Western blot of endogenous protein","pmids":["25436799"],"confidence":"Medium","gaps":["Physiological contexts where miR-611 is expressed not mapped","Clinical/phenotypic consequence of the SNP not established","Single lab"]},{"year":2019,"claim":"Whether surface mesothelin could be exploited for targeted therapy was being tested; a MSLN-directed thorium-227 alpha conjugate established that antigen binding can deliver cytotoxic radiation causing DNA double-strand breaks and tumor regression.","evidence":"In vitro binding, γH2AX/apoptosis assays, and multiple cancer xenograft efficacy and biodistribution studies","pmids":["31064781"],"confidence":"Medium","gaps":["Demonstrates targeting, not endogenous mesothelin signaling","Off-target/normal-tissue toxicity not the focus","Single developer"]},{"year":2021,"claim":"The physiological function of mesothelin outside cancer was undefined; knockout mouse studies established a MSLN-Thy1 signaling complex driving fibroblast activation and a pro-fibrotic role across multiple organs.","evidence":"Msln-/- and Thy1-/- mice across BDL, bleomycin, and UUO fibrosis models with anti-MSLN immunotoxin treatment","pmids":["34253615"],"confidence":"High","gaps":["Biochemical nature of the MSLN-Thy1 interaction not structurally resolved","Downstream signaling driving fibroblast activation not fully defined","Human fibrosis relevance inferred from mouse models"]},{"year":2021,"claim":"How mesothelin links to oncogenic signaling was unclear; metformin-induced MSLN downregulation reducing IL-6/STAT3 activity placed mesothelin upstream of this axis in ovarian cancer.","evidence":"Metformin treatment with knockdown/overexpression and Western blot of IL-6/STAT3, VEGF, TGFβ1 in ovarian cancer cells","pmids":["34238029"],"confidence":"Low","gaps":["No direct reconstitution of a MSLN–IL-6/STAT3 interaction","Pathway placement inferred from expression changes after drug treatment","Single lab, correlative"]},{"year":null,"claim":"The molecular receptor/effector mechanism by which mesothelin transduces signals to drive both fibroblast activation and tumor cell phenotypes remains undefined.","evidence":"","pmids":[],"confidence":"Medium","gaps":["Structure of the MSLN-Thy1 complex unknown","Direct downstream signaling cascade not mapped","Whether fibrosis and cancer roles share one mechanism unresolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[2]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[2,6]}],"pathway":[],"complexes":[],"partners":["THY1","TEAD1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q13421","full_name":"Mesothelin","aliases":["CAK1 antigen","Pre-pro-megakaryocyte-potentiating factor"],"length_aa":630,"mass_kda":69.0,"function":"Membrane-anchored forms may play a role in cellular adhesion Megakaryocyte-potentiating factor (MPF) potentiates megakaryocyte colony formation in vitro","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/Q13421/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MSLN","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/MSLN","total_profiled":1310},"omim":[{"mim_id":"601051","title":"MESOTHELIN; MSLN","url":"https://www.omim.org/entry/601051"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Vesicles","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"adipose tissue","ntpm":89.2},{"tissue":"fallopian tube","ntpm":261.5}],"url":"https://www.proteinatlas.org/search/MSLN"},"hgnc":{"alias_symbol":["CAK1","MPF"],"prev_symbol":[]},"alphafold":{"accession":"Q13421","domains":[{"cath_id":"-","chopping":"59-275","consensus_level":"medium","plddt":87.8198,"start":59,"end":275},{"cath_id":"-","chopping":"330-432","consensus_level":"medium","plddt":84.0996,"start":330,"end":432}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q13421","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q13421-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q13421-F1-predicted_aligned_error_v6.png","plddt_mean":78.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MSLN","jax_strain_url":"https://www.jax.org/strain/search?query=MSLN"},"sequence":{"accession":"Q13421","fasta_url":"https://rest.uniprot.org/uniprotkb/Q13421.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q13421/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q13421"}},"corpus_meta":[{"pmid":"3293802","id":"PMC_3293802","title":"The Xenopus cdc2 protein is a component of MPF, a cytoplasmic regulator of mitosis.","date":"1988","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/3293802","citation_count":748,"is_preprint":false},{"pmid":"2538242","id":"PMC_2538242","title":"Cdc2 protein kinase is complexed with both cyclin A and B: evidence for proteolytic inactivation of MPF.","date":"1989","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/2538242","citation_count":627,"is_preprint":false},{"pmid":"8428594","id":"PMC_8428594","title":"Phosphorylation and activation of human cdc25-C by cdc2--cyclin B and its involvement in the self-amplification of MPF at mitosis.","date":"1993","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/8428594","citation_count":586,"is_preprint":false},{"pmid":"2844417","id":"PMC_2844417","title":"cdc2 is a component of the M phase-specific histone H1 kinase: evidence for identity with 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in the length of the meiotic stages.","date":"2001","source":"Animal reproduction science","url":"https://pubmed.ncbi.nlm.nih.gov/11348780","citation_count":61,"is_preprint":false},{"pmid":"1825521","id":"PMC_1825521","title":"A model for the adjustment of the mitotic clock by cyclin and MPF levels.","date":"1991","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/1825521","citation_count":61,"is_preprint":false},{"pmid":"16921258","id":"PMC_16921258","title":"Cyclin B dissociation from CDK1 precedes its degradation upon MPF inactivation in mitotic extracts of Xenopus laevis embryos.","date":"2006","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/16921258","citation_count":58,"is_preprint":false},{"pmid":"25712366","id":"PMC_25712366","title":"Entry into mitosis: a solution to the decades-long enigma of MPF.","date":"2015","source":"Chromosoma","url":"https://pubmed.ncbi.nlm.nih.gov/25712366","citation_count":56,"is_preprint":false},{"pmid":"17909009","id":"PMC_17909009","title":"High cancer-specific expression of mesothelin (MSLN) is attributable to an upstream enhancer containing a transcription enhancer factor dependent MCAT motif.","date":"2007","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/17909009","citation_count":56,"is_preprint":false},{"pmid":"10491252","id":"PMC_10491252","title":"Activation of MPF at meiosis reinitiation in starfish oocytes.","date":"1999","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/10491252","citation_count":56,"is_preprint":false},{"pmid":"3044877","id":"PMC_3044877","title":"Monoclonal antibodies specific for thiophosphorylated proteins recognize Xenopus MPF.","date":"1988","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/3044877","citation_count":56,"is_preprint":false},{"pmid":"34803504","id":"PMC_34803504","title":"Tandem CAR-T cells targeting FOLR1 and MSLN enhance the antitumor effects in ovarian cancer.","date":"2021","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34803504","citation_count":54,"is_preprint":false},{"pmid":"9806800","id":"PMC_9806800","title":"MPF amplification in Xenopus oocyte extracts depends on a two-step activation of cdc25 phosphatase.","date":"1998","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/9806800","citation_count":54,"is_preprint":false},{"pmid":"9665339","id":"PMC_9665339","title":"Maturation-promoting factor (MPF) and mitogen activated protein kinase (MAPK) expression in relation to oocyte competence for in-vitro maturation in the mare.","date":"1998","source":"Molecular human reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/9665339","citation_count":52,"is_preprint":false},{"pmid":"1361167","id":"PMC_1361167","title":"A novel homo-oligomeric protein responsible for an MPF-dependent microtubule-severing activity.","date":"1992","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/1361167","citation_count":51,"is_preprint":false},{"pmid":"9427686","id":"PMC_9427686","title":"A propagated wave of MPF activation accompanies surface contraction waves at first mitosis in Xenopus.","date":"1998","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/9427686","citation_count":50,"is_preprint":false},{"pmid":"36746513","id":"PMC_36746513","title":"Tuned activation of MSLN-CAR T cells induces superior antitumor responses in ovarian cancer models.","date":"2023","source":"Journal for immunotherapy of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/36746513","citation_count":49,"is_preprint":false},{"pmid":"8381222","id":"PMC_8381222","title":"p21ras-induced meiotic maturation of Xenopus oocytes in the absence of protein synthesis: MPF activation is preceded by activation of MAP and S6 kinases.","date":"1993","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/8381222","citation_count":48,"is_preprint":false},{"pmid":"29643273","id":"PMC_29643273","title":"MPF-based meiotic cell cycle control: Half a century of lessons from starfish oocytes.","date":"2018","source":"Proceedings of the Japan Academy. Series B, Physical and biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/29643273","citation_count":45,"is_preprint":false},{"pmid":"12756171","id":"PMC_12756171","title":"Endoplasmic reticulum reorganizations and Ca2+ signaling in maturing and fertilized oocytes of marine protostome worms: the roles of MAPKs and MPF.","date":"2003","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/12756171","citation_count":45,"is_preprint":false},{"pmid":"11933170","id":"PMC_11933170","title":"Association of MPF, MAPK, and nuclear progression dynamics during activation of young and aged bovine oocytes.","date":"2002","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/11933170","citation_count":41,"is_preprint":false},{"pmid":"17014901","id":"PMC_17014901","title":"Effect of oocyte diameter on meiotic competence, embryo development, p34 (cdc2) expression and MPF activity in prepubertal goat oocytes.","date":"2006","source":"Theriogenology","url":"https://pubmed.ncbi.nlm.nih.gov/17014901","citation_count":40,"is_preprint":false},{"pmid":"35898704","id":"PMC_35898704","title":"Trogocytosis and fratricide killing impede MSLN-directed CAR T cell functionality.","date":"2022","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/35898704","citation_count":37,"is_preprint":false},{"pmid":"30260046","id":"PMC_30260046","title":"MSLN (Mesothelin), ANTXR1 (TEM8), and MUC3A are the potent antigenic targets for CAR T cell therapy of gastric adenocarcinoma.","date":"2018","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30260046","citation_count":37,"is_preprint":false},{"pmid":"15457541","id":"PMC_15457541","title":"Association of female aging with decreased parthenogenetic activation, raised MPF, and MAPKs activities and reduced levels of glutathione S-transferases activity and thiols in mouse oocytes.","date":"2004","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/15457541","citation_count":37,"is_preprint":false},{"pmid":"12482715","id":"PMC_12482715","title":"Discrimination of the roles of MPF and MAP kinase in morphological changes that occur during oocyte maturation.","date":"2002","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/12482715","citation_count":36,"is_preprint":false},{"pmid":"9520324","id":"PMC_9520324","title":"Changes in cyclin B during oocyte maturation and early embryonic cell cycle in the newt, Cynops pyrrhogaster: requirement of germinal vesicle for MPF activation.","date":"1998","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/9520324","citation_count":36,"is_preprint":false},{"pmid":"23028227","id":"PMC_23028227","title":"A MSLN-targeted multifunctional nanoimmunoliposome for MRI and targeting therapy in pancreatic cancer.","date":"2012","source":"International journal of nanomedicine","url":"https://pubmed.ncbi.nlm.nih.gov/23028227","citation_count":35,"is_preprint":false},{"pmid":"14985258","id":"PMC_14985258","title":"Polo-like kinase confers MPF autoamplification competence to growing Xenopus oocytes.","date":"2004","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/14985258","citation_count":35,"is_preprint":false},{"pmid":"16628004","id":"PMC_16628004","title":"M-phase MELK activity is regulated by MPF and MAPK.","date":"2006","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/16628004","citation_count":35,"is_preprint":false},{"pmid":"7576618","id":"PMC_7576618","title":"UV irradiation of chromosomal DNA and its effect upon MPF and meiosis in mammalian oocytes.","date":"1995","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/7576618","citation_count":35,"is_preprint":false},{"pmid":"24167356","id":"PMC_24167356","title":"Comparison of the diagnostic accuracy of the MSLN gene products, mesothelin and megakaryocyte potentiating factor, as biomarkers for mesothelioma in pleural effusions and serum.","date":"2013","source":"Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/24167356","citation_count":34,"is_preprint":false},{"pmid":"37280838","id":"PMC_37280838","title":"The Role of \"Cytostatic Factor (CSF)\" in the Control of Oocyte Cell Cycles: A Summary of 20 Years of Study: (CSF/MPF/Cell Cycle/Metaphase Arrest).","date":"1991","source":"Development, growth & differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/37280838","citation_count":34,"is_preprint":false},{"pmid":"7735115","id":"PMC_7735115","title":"Microinjection of Cdc25 protein phosphatase into Xenopus prophase oocyte activates MPF and arrests meiosis at metaphase I.","date":"1994","source":"Biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/7735115","citation_count":34,"is_preprint":false},{"pmid":"34238029","id":"PMC_34238029","title":"Metformin Antagonizes Ovarian Cancer Cells Malignancy Through MSLN Mediated IL-6/STAT3 Signaling.","date":"2021","source":"Cell transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/34238029","citation_count":33,"is_preprint":false},{"pmid":"10973988","id":"PMC_10973988","title":"Nuclei and microtubule asters stimulate maturation/M phase promoting factor (MPF) activation in Xenopus eggs and egg cytoplasmic extracts.","date":"2000","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/10973988","citation_count":33,"is_preprint":false},{"pmid":"23333317","id":"PMC_23333317","title":"Removal of centrosomal PP1 by NIMA kinase unlocks the MPF feedback loop to promote mitotic commitment in S. pombe.","date":"2013","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/23333317","citation_count":32,"is_preprint":false},{"pmid":"9291480","id":"PMC_9291480","title":"Protein kinase C is required for the disappearance of MPF upon artificial activation in mouse eggs.","date":"1997","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/9291480","citation_count":31,"is_preprint":false},{"pmid":"15457512","id":"PMC_15457512","title":"Protein synthesis and mRNA storage in cattle oocytes maintained under meiotic block by roscovitine inhibition of MPF activity.","date":"2004","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/15457512","citation_count":29,"is_preprint":false},{"pmid":"10683314","id":"PMC_10683314","title":"Structure of the Mesothelin/MPF gene and characterization of its promoter.","date":"2000","source":"Molecular cell biology research communications : MCBRC","url":"https://pubmed.ncbi.nlm.nih.gov/10683314","citation_count":27,"is_preprint":false},{"pmid":"17443687","id":"PMC_17443687","title":"Zinc regulates the ability of Cdc25C to activate MPF/cdk1.","date":"2007","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/17443687","citation_count":27,"is_preprint":false},{"pmid":"8625296","id":"PMC_8625296","title":"Radiation with 1 Gy prevents the activation of the mitotic inducers mitosis-promoting factor (MPF) and cdc25-C in HeLa cells.","date":"1996","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/8625296","citation_count":27,"is_preprint":false},{"pmid":"34253615","id":"PMC_34253615","title":"Immunotherapy-based targeting of MSLN+ activated portal fibroblasts is a strategy for treatment of cholestatic liver fibrosis.","date":"2021","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/34253615","citation_count":26,"is_preprint":false},{"pmid":"2147396","id":"PMC_2147396","title":"MPF and cyclin: modelling of the cell cycle minimum oscillator.","date":"1990","source":"Bio Systems","url":"https://pubmed.ncbi.nlm.nih.gov/2147396","citation_count":26,"is_preprint":false},{"pmid":"31887595","id":"PMC_31887595","title":"Microcystin-LR promotes zebrafish (Danio rerio) oocyte (in vivo) maturation by activating ERK1/2-MPF signaling pathways, and cAMP is involved in this process.","date":"2019","source":"Environmental pollution (Barking, Essex : 1987)","url":"https://pubmed.ncbi.nlm.nih.gov/31887595","citation_count":26,"is_preprint":false},{"pmid":"15118243","id":"PMC_15118243","title":"Activation with ethanol improves embryo development of ICSI-derived oocytes by regulation of kinetics of MPF activity.","date":"2004","source":"The Journal of reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/15118243","citation_count":26,"is_preprint":false},{"pmid":"20573372","id":"PMC_20573372","title":"Mesothelin (MSLN) promoter is hypomethylated in malignant mesothelioma, but its expression is not associated with methylation status of the promoter.","date":"2010","source":"Human pathology","url":"https://pubmed.ncbi.nlm.nih.gov/20573372","citation_count":25,"is_preprint":false},{"pmid":"24465798","id":"PMC_24465798","title":"MSLN gene silencing has an anti-malignant effect on cell lines overexpressing mesothelin deriving from malignant pleural mesothelioma.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24465798","citation_count":24,"is_preprint":false},{"pmid":"21775819","id":"PMC_21775819","title":"The relationship between tumor MSLN methylation and serum mesothelin (SMRP) in mesothelioma.","date":"2011","source":"Epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/21775819","citation_count":24,"is_preprint":false},{"pmid":"12363437","id":"PMC_12363437","title":"In vitro maturation of bovine oocytes requires polyadenylation of mRNAs coding proteins for chromatin condensation, spindle assembly, MPF and MAP kinase activation.","date":"2002","source":"Animal reproduction science","url":"https://pubmed.ncbi.nlm.nih.gov/12363437","citation_count":23,"is_preprint":false},{"pmid":"2838346","id":"PMC_2838346","title":"The cell cycle can occur in starfish oocytes and embryos without the production of transferable MPF (maturation-promoting factor).","date":"1988","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/2838346","citation_count":23,"is_preprint":false},{"pmid":"23781026","id":"PMC_23781026","title":"The phosphorylation of ARPP19 by Greatwall renders the auto-amplification of MPF independently of PKA in Xenopus oocytes.","date":"2013","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/23781026","citation_count":23,"is_preprint":false},{"pmid":"11977986","id":"PMC_11977986","title":"Characterization of MPF and MAPK activities during meiotic maturation of Xenopus tropicalis oocytes.","date":"2002","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/11977986","citation_count":22,"is_preprint":false},{"pmid":"15278898","id":"PMC_15278898","title":"Timing of Plk1 and MPF activation during porcine oocyte maturation.","date":"2004","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/15278898","citation_count":21,"is_preprint":false},{"pmid":"12713427","id":"PMC_12713427","title":"Independent activation of MAP kinase and MPF during the initiation of meiotic maturation in pig oocytes.","date":"2003","source":"Reproduction (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/12713427","citation_count":20,"is_preprint":false},{"pmid":"25436799","id":"PMC_25436799","title":"A common polymorphism within MSLN affects miR-611 binding site and soluble mesothelin levels in healthy people.","date":"2014","source":"Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/25436799","citation_count":20,"is_preprint":false},{"pmid":"3068110","id":"PMC_3068110","title":"Does autocatalytic amplification of maturation-promoting factor (MPF) exist in mammalian oocytes?","date":"1988","source":"Gamete research","url":"https://pubmed.ncbi.nlm.nih.gov/3068110","citation_count":20,"is_preprint":false},{"pmid":"19292671","id":"PMC_19292671","title":"Characterization and in vitro control of MPF activity in zebrafish eggs.","date":"2009","source":"Zebrafish","url":"https://pubmed.ncbi.nlm.nih.gov/19292671","citation_count":18,"is_preprint":false},{"pmid":"8398135","id":"PMC_8398135","title":"Mitogen-activated protein kinase (MAP kinase) activation in Xenopus oocytes: roles of MPF and protein synthesis.","date":"1993","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/8398135","citation_count":18,"is_preprint":false},{"pmid":"15107242","id":"PMC_15107242","title":"M-phase promoting factor (MPF) and mitogen activated protein kinases (MAPK) activities of domestic cat oocytes matured in vitro and in vivo.","date":"2004","source":"Cloning and stem cells","url":"https://pubmed.ncbi.nlm.nih.gov/15107242","citation_count":17,"is_preprint":false},{"pmid":"7541376","id":"PMC_7541376","title":"Brefeldin A provokes indirect activation of cdc2 kinase (MPF) in Xenopus oocytes, resulting in meiotic cell division.","date":"1995","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/7541376","citation_count":17,"is_preprint":false},{"pmid":"7962177","id":"PMC_7962177","title":"Progesterone but not ras requires MPF for in vivo activation of MAPK and S6 KII: MAPK is an essential conexion point of both signaling pathways.","date":"1994","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/7962177","citation_count":17,"is_preprint":false},{"pmid":"3539598","id":"PMC_3539598","title":"Partial purification of the maturation-promoting factor MPF from unfertilized eggs of Xenopus laevis.","date":"1986","source":"European journal of biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/3539598","citation_count":17,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":46678,"output_tokens":2551,"usd":0.08915,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9019,"output_tokens":2666,"usd":0.055872,"stage2_stop_reason":"end_turn"},"total_usd":0.145022,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"Looking at these papers, I need to classify them carefully. The query gene is MSLN (Mesothelin), with alias MPF. However, the vast majority of these papers are about \"MPF\" = Maturation-Promoting Factor (the cdc2/cyclin B complex), which is a completely different biological entity from MSLN/Mesothelin. Only a few papers actually concern the MSLN gene product (mesothelin protein).\n\nLet me identify the papers genuinely about MSLN (mesothelin):\n- Paper [28]: MSLN-targeted thorium-227 conjugate for cancer therapy\n- Paper [46]: MSLN gene transcription regulation via TEF-1/TEAD1 and CanScript enhancer\n- Paper [49]: FOLR1 and MSLN CAR-T cells for ovarian cancer\n- Paper [54]: MSLN-CAR T cells with ITAM mutations\n- Paper [60]: Trogocytosis in MSLN-CAR T cells\n- Paper [61]: MSLN as CAR T target in gastric cancer (bioinformatics, no mechanism)\n- Paper [65]: MSLN-targeted nanoimmunoliposome for pancreatic cancer\n- Paper [69]: MSLN gene products (mesothelin and MPF/megakaryocyte potentiating factor) as biomarkers\n- Paper [72]: Metformin/MSLN/IL-6/STAT3 signaling in ovarian cancer\n- Paper [77]: MSLN gene structure and promoter characterization\n- Paper [80]: MSLN role in fibroblast activation and fibrosis\n- Paper [84]: MSLN promoter methylation\n- Paper [85]: MSLN silencing effects on mesothelioma cells\n- Paper [86]: MSLN methylation and SMRP levels\n- Paper [93]: MSLN SNP affecting miR-611 binding\n\nAll other papers (the bulk) are about MPF = Maturation-Promoting Factor (cdc2/cyclin B), which is a symbol collision with MSLN's alias \"MPF\" = Megakaryocyte Potentiating Factor. These are completely unrelated proteins.\n\nNow extracting mechanistic discoveries from MSLN papers:\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2000,\n      \"finding\": \"The MSLN gene spans 8 kb of human chromosome 16, is encoded by 15 exons with an 1884-bp open reading frame, and its promoter lacks a TATA box and SP1 sites. Transient transfection assays localized a minimal constitutive promoter to a 317-bp region and identified tissue-specific enhancer elements upstream of this minimal promoter that activate transcription in a position- and orientation-independent manner.\",\n      \"method\": \"Transient transfection analysis, promoter deletion constructs, genomic sequencing\",\n      \"journal\": \"Molecular cell biology research communications : MCBRC\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct promoter functional dissection with deletion constructs and transfection assays in a single study; no replication by independent lab\",\n      \"pmids\": [\"10683314\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"An 18-bp upstream enhancer element called 'CanScript' drives cancer-specific MSLN overexpression. The CanScript contains two functional sites: an Sp1-like site and an MCAT element. Gel retardation and chromatin immunoprecipitation assays showed that transcription enhancer factor TEF-1 (TEAD1) binds the MCAT element both in vitro and in vivo, and TEF-1 knockdown reduced MSLN protein overexpression in cancer cells.\",\n      \"method\": \"Promoter deletion/mutation analysis, gel retardation (EMSA), chromatin immunoprecipitation (ChIP), TEF-1 siRNA knockdown, transient transfection\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (EMSA, ChIP, mutagenesis, knockdown) in one study establishing TEF-1/TEAD1 as direct transcriptional activator of MSLN via CanScript MCAT element\",\n      \"pmids\": [\"17909009\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MSLN knockout mice (Msln-/-) are protected from cholestatic liver fibrosis (Mdr2-deficient model), bleomycin-induced lung fibrosis, and UUO-induced kidney fibrosis, while Thy1-/- mice are more susceptible, indicating that a MSLN-Thy1 signaling complex is critical for tissue fibroblast activation. Targeting MSLN+ activated portal fibroblasts with anti-MSLN immunotoxins reduced collagen deposition in bile duct ligation-injured mice.\",\n      \"method\": \"Genetic knockout mouse models (Msln-/-, Thy1-/-), multiple fibrosis models (BDL, bleomycin, UUO), anti-MSLN immunotoxin treatment, collagen deposition assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function genetic models replicated across three distinct fibrosis models with defined cellular phenotype; functional rescue with immunotoxin\",\n      \"pmids\": [\"34253615\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Metformin treatment in ovarian cancer cells reduced MSLN expression, which in turn downregulated IL-6/STAT3 signaling activity and subsequently reduced VEGF and TGFβ1 expression, leading to inhibition of cell growth, migration, and stemness.\",\n      \"method\": \"Cell-based assays (proliferation, migration, apoptosis), molecular knockdown/overexpression, Western blot for pathway components\",\n      \"journal\": \"Cell transplantation\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement based on expression changes after metformin treatment; no direct reconstitution of MSLN–IL-6/STAT3 interaction\",\n      \"pmids\": [\"34238029\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Transient siRNA-mediated MSLN knockdown in MSLN-overexpressing mesothelioma cells (Mero-14) caused decreased proliferation, reduced invasive capacity, and reduced sphere formation; combined MSLN-siRNA with cisplatin triggered increased apoptosis and decreased proliferation compared to either agent alone.\",\n      \"method\": \"siRNA knockdown, proliferation assay, invasion assay, sphere formation assay, apoptosis assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — clean loss-of-function with defined cellular phenotypes (proliferation, invasion, sphere formation) across multiple readouts in a single study; replicated previous findings from other cell lines\",\n      \"pmids\": [\"24465798\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"A common SNP (rs1057147) in the 3' UTR of MSLN creates a binding site for miR-611. Dual-luciferase reporter assays showed lower reporter activity with the G allele vs. A allele; miR-611 mimic reduced reporter activity only of the G-allele construct. Transfection of miR-611 into mesothelioma cells (Mero-14) significantly reduced endogenous MSLN protein levels, indicating post-transcriptional regulation of MSLN by miR-611 in a genotype-dependent manner.\",\n      \"method\": \"Dual-luciferase reporter assay with allele-specific constructs, miRNA mimic transfection, Western blot for endogenous MSLN protein\",\n      \"journal\": \"Journal of thoracic oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional reporter assay plus endogenous protein knockdown by miRNA in single lab; two orthogonal methods establish miR-611 as post-transcriptional regulator of MSLN\",\n      \"pmids\": [\"25436799\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MSLN-targeted thorium-227 conjugate (BAY 2287411) binds MSLN-expressing tumor cells and, upon delivery of alpha-particle radiation, induces DNA double-strand breaks, apoptotic markers, and oxidative stress, leading to reduced cellular viability and significant antitumor efficacy in vivo in multiple cancer xenograft models.\",\n      \"method\": \"In vitro binding assay, DNA damage markers (γH2AX), apoptosis assays, in vivo xenograft models (breast, colorectal, lung, ovarian, pancreatic), biodistribution studies\",\n      \"journal\": \"Clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple in vitro and in vivo models with defined mechanism of action (MSLN-targeted alpha-radiation causing DSBs); single lab but multiple orthogonal readouts\",\n      \"pmids\": [\"31064781\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MSLN encodes mesothelin, a GPI-anchored membrane glycoprotein whose cancer-specific overexpression is driven by TEF-1/TEAD1 binding to an MCAT-containing enhancer element (CanScript) in its promoter; MSLN expression is also post-transcriptionally regulated by miR-611 in a SNP-dependent manner; functionally, MSLN forms a signaling complex with Thy1 that promotes tissue fibroblast activation across multiple organs, and loss of MSLN protects mice from fibrosis, while MSLN knockdown in cancer cells reduces proliferation, invasion, and stemness, partly through modulation of IL-6/STAT3 signaling.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MSLN encodes mesothelin, a cell-surface glycoprotein whose cancer-specific overexpression is driven transcriptionally by the enhancer factor TEF-1/TEAD1, which binds the MCAT element within an 18-bp upstream 'CanScript' enhancer; TEF-1 knockdown reduces mesothelin overexpression in cancer cells [#1]. The MSLN promoter lacks TATA and SP1 boxes and depends on a minimal constitutive region plus position- and orientation-independent tissue-specific enhancers [#0]. Mesothelin expression is additionally tuned post-transcriptionally: a common 3' UTR SNP (rs1057147) creates an allele-specific miR-611 binding site, and miR-611 reduces endogenous mesothelin protein in a genotype-dependent manner [#5]. Functionally, mesothelin acts in a MSLN-Thy1 signaling axis that drives tissue fibroblast activation; Msln-knockout mice are protected from cholestatic liver, bleomycin lung, and UUO kidney fibrosis, and targeting MSLN+ portal fibroblasts reduces collagen deposition [#2]. In cancer cells, loss of mesothelin diminishes proliferation, invasion, and sphere-forming stemness [#4]. Its surface accessibility makes mesothelin a tractable therapeutic target, exemplified by a MSLN-directed thorium-227 alpha-particle conjugate that induces DNA double-strand breaks and antitumor efficacy across xenograft models [#6].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Defining the architecture of the MSLN promoter was needed to understand how the gene is transcriptionally controlled; this established a TATA-less minimal promoter governed by separable upstream tissue-specific enhancers.\",\n      \"evidence\": \"Promoter deletion constructs and transient transfection assays with genomic sequencing\",\n      \"pmids\": [\"10683314\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Identity of the trans-acting factors binding the enhancers not resolved\", \"No link to cancer-specific expression at this stage\", \"Single study without independent replication\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"It was unknown which factor drove cancer-selective MSLN overexpression; identification of TEF-1/TEAD1 binding the MCAT element of the CanScript enhancer established a direct transcriptional activator of MSLN in cancer.\",\n      \"evidence\": \"EMSA, ChIP, promoter mutagenesis, and TEF-1 siRNA knockdown in cancer cells\",\n      \"pmids\": [\"17909009\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream signals controlling TEF-1 activity at this enhancer unknown\", \"Role of the adjacent Sp1-like site not fully dissected\", \"Whether the same mechanism operates in non-malignant tissue not addressed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Whether mesothelin had a cell-autonomous role in tumor biology was unclear; siRNA knockdown in mesothelioma cells established that mesothelin supports proliferation, invasion, and stemness and sensitizes cells to cisplatin.\",\n      \"evidence\": \"siRNA knockdown with proliferation, invasion, sphere formation, and apoptosis assays in Mero-14 cells\",\n      \"pmids\": [\"24465798\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular effectors downstream of mesothelin not defined here\", \"Transient knockdown only\", \"In vivo relevance not tested\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"The basis for inter-individual variation in mesothelin levels was unknown; a 3' UTR SNP creating an allele-specific miR-611 site established post-transcriptional, genotype-dependent regulation of MSLN.\",\n      \"evidence\": \"Allele-specific dual-luciferase reporters, miR-611 mimic transfection, and Western blot of endogenous protein\",\n      \"pmids\": [\"25436799\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological contexts where miR-611 is expressed not mapped\", \"Clinical/phenotypic consequence of the SNP not established\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Whether surface mesothelin could be exploited for targeted therapy was being tested; a MSLN-directed thorium-227 alpha conjugate established that antigen binding can deliver cytotoxic radiation causing DNA double-strand breaks and tumor regression.\",\n      \"evidence\": \"In vitro binding, \\u03b3H2AX/apoptosis assays, and multiple cancer xenograft efficacy and biodistribution studies\",\n      \"pmids\": [\"31064781\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Demonstrates targeting, not endogenous mesothelin signaling\", \"Off-target/normal-tissue toxicity not the focus\", \"Single developer\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"The physiological function of mesothelin outside cancer was undefined; knockout mouse studies established a MSLN-Thy1 signaling complex driving fibroblast activation and a pro-fibrotic role across multiple organs.\",\n      \"evidence\": \"Msln-/- and Thy1-/- mice across BDL, bleomycin, and UUO fibrosis models with anti-MSLN immunotoxin treatment\",\n      \"pmids\": [\"34253615\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biochemical nature of the MSLN-Thy1 interaction not structurally resolved\", \"Downstream signaling driving fibroblast activation not fully defined\", \"Human fibrosis relevance inferred from mouse models\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"How mesothelin links to oncogenic signaling was unclear; metformin-induced MSLN downregulation reducing IL-6/STAT3 activity placed mesothelin upstream of this axis in ovarian cancer.\",\n      \"evidence\": \"Metformin treatment with knockdown/overexpression and Western blot of IL-6/STAT3, VEGF, TGF\\u03b21 in ovarian cancer cells\",\n      \"pmids\": [\"34238029\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct reconstitution of a MSLN\\u2013IL-6/STAT3 interaction\", \"Pathway placement inferred from expression changes after drug treatment\", \"Single lab, correlative\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The molecular receptor/effector mechanism by which mesothelin transduces signals to drive both fibroblast activation and tumor cell phenotypes remains undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structure of the MSLN-Thy1 complex unknown\", \"Direct downstream signaling cascade not mapped\", \"Whether fibrosis and cancer roles share one mechanism unresolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [2, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0001618\", \"supporting_discovery_ids\": []}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"THY1\",\n      \"TEAD1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}