{"gene":"SHISA3","run_date":"2026-06-10T07:46:31","timeline":{"discoveries":[{"year":2018,"finding":"SHISA3 suppresses NPC cell invasion and metastasis by impeding TRIM21-mediated ubiquitination and degradation of SGSM1, thereby inhibiting MAPK pathway activation. Silencing SGSM1 abrogated the inhibitory effect of SHISA3 on cell migration and invasion, placing SHISA3 upstream of TRIM21-SGSM1-MAPK axis.","method":"In vitro invasion/migration assays, in vivo metastasis models, co-immunoprecipitation, ubiquitination assays, SGSM1 knockdown rescue experiments","journal":"Cancer Research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal functional epistasis (SGSM1 KD rescue), IP-based interaction, in vitro and in vivo phenotypes, single lab with multiple orthogonal methods","pmids":["30573520"],"is_preprint":false},{"year":2014,"finding":"SHISA3 accelerates β-catenin degradation to suppress WNT signaling, thereby inhibiting tumorigenesis and invasion in lung cancer cells in vitro and reducing tumor growth and metastasis in mouse models.","method":"Microarray/pathway analysis, in vitro invasion/migration assays, subcutaneous and orthotopic mouse tumor models, β-catenin degradation assays","journal":"American Journal of Respiratory and Critical Care Medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro and in vivo functional assays with mechanistic readout (β-catenin degradation), single lab, multiple methods","pmids":["25036006"],"is_preprint":false},{"year":2019,"finding":"SHISA3 interacts with FGFR1 and FGFR3 to inhibit AKT/mTOR signaling, thereby suppressing cancer stem cell properties and reversing EGFR-TKI resistance in lung adenocarcinoma.","method":"Immunoprecipitation, Western blot, sphere formation assay, Transwell assay, CCK8, in vivo NOD-SCID mouse model, pharmacological inhibition","journal":"Journal of Experimental & Clinical Cancer Research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-immunoprecipitation demonstrating FGFR1/3 interaction, downstream signaling (AKT/mTOR) by Western blot, in vivo validation, single lab","pmids":["31801598"],"is_preprint":false},{"year":2019,"finding":"SHISA3 blocks maturation and transportation of Frizzled receptors to the cell surface, inhibiting Wnt/β-catenin signaling; adenovirus-mediated overexpression of Shisa3 significantly inhibited Wnt3a-induced nuclear translocation of β-catenin and mRNA expression of Wnt target gene Axin2 in osteoblasts. Shisa3 knockout mice showed no bone phenotype, suggesting functional redundancy with other Shisa family members.","method":"Adenovirus-mediated gene transfer, β-catenin nuclear translocation assay, Axin2 mRNA expression, micro-CT, Shisa3 KO mouse phenotyping","journal":"Journal of Bone and Mineral Metabolism","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct functional assay (Wnt3a-induced β-catenin translocation inhibition) plus KO mouse phenotyping, single lab, multiple methods","pmids":["31222549"],"is_preprint":false},{"year":2019,"finding":"SHISA3 expression in the heart is regulated by KLF15-Wnt-dependent transcriptional networks; KLF15 represses Wnt-dependent transcriptional signaling postnatally, and loss of KLF15 leads to Wnt-dependent SHISA3 upregulation. SHISA3 is primarily expressed in vascular cells (VCs) in fetal hearts and during pathological remodeling, conserved in mouse and human models.","method":"Transcriptomic bioinformatics, Klf15 KO mouse phenotyping, Wnt-signaling modulated hearts, pressure overload and myocardial ischemia models, human KLF15 KO embryonic stem cells, engineered human myocardium","journal":"Journal of the American College of Cardiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO models plus human stem cell validation, multiple orthogonal methods, single lab","pmids":["31582141"],"is_preprint":false},{"year":2024,"finding":"SHISA3 expression in macrophages is induced by DAMPs/PAMPs via NF-κB transcription factors. Reciprocally, SHISA3 forms a complex with HSPA8 to activate NF-κB signaling, maintaining M1 polarization of macrophages, increasing phagocytosis and antigen presentation, and promoting CD8+ T cell-mediated antitumor immunity. Shisa3 knockout abolished the antitumor efficacy of TLR4 agonist MPLA combined with PD-1 antibody.","method":"mRNA delivery, co-immunoprecipitation (SHISA3-HSPA8 complex), NF-κB signaling assays, phagocytosis and antigen presentation assays, Shisa3 KO mouse experiments, combination immunotherapy in vivo","journal":"Advanced Science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-immunoprecipitation of SHISA3-HSPA8, NF-κB pathway validation, KO functional rescue in vivo, single lab, multiple orthogonal methods","pmids":["39054639"],"is_preprint":false},{"year":2020,"finding":"Ectopic expression of SHISA3 in breast cancer cell lines (MDA-MB231, MCF-7) significantly reduced proliferation and migration. SHISA3 acts as an antagonist of Wnt/β-catenin signaling and its promoter is epigenetically silenced by hypermethylation; 5-aza-2'-deoxycytidine treatment restored SHISA3 expression.","method":"Ectopic expression, proliferation and migration assays, methylation-specific PCR, 5-aza-2'-deoxycytidine demethylation treatment","journal":"PLoS One","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — functional assays (proliferation/migration) with ectopic expression, demethylation rescue, single lab, multiple methods but no direct mechanistic protein interaction shown","pmids":["32692756"],"is_preprint":false}],"current_model":"SHISA3 is a tumor suppressor and Wnt/β-catenin antagonist that blocks Frizzled receptor maturation/transport to inhibit Wnt signaling and accelerate β-catenin degradation; it also suppresses invasion and metastasis via a TRIM21-SGSM1-MAPK axis, interacts with FGFR1/3 to inhibit AKT/mTOR signaling and cancer stem cell properties, and in macrophages forms a complex with HSPA8 to activate NF-κB signaling and maintain M1 polarization, with its expression regulated by KLF15-Wnt transcriptional networks in the heart and by DAMPs/PAMPs-induced NF-κB signaling in immune cells."},"narrative":{"mechanistic_narrative":"SHISA3 is a tumor-suppressive antagonist of Wnt/β-catenin signaling that also engages receptor tyrosine kinase and innate immune pathways to restrain tumor growth, invasion, and metastasis [PMID:25036006, PMID:31222549]. It inhibits Wnt signaling by blocking maturation and transport of Frizzled receptors to the cell surface, preventing Wnt3a-induced nuclear translocation of β-catenin and expression of the target gene Axin2, and by accelerating β-catenin degradation [PMID:25036006, PMID:31222549]. In carcinoma cells SHISA3 suppresses invasion and metastasis by impeding TRIM21-mediated ubiquitination and degradation of SGSM1, thereby dampening MAPK pathway activation [PMID:30573520], and it interacts with FGFR1 and FGFR3 to inhibit AKT/mTOR signaling, suppress cancer stem cell properties, and reverse EGFR-TKI resistance [PMID:31801598]. Its expression is silenced by promoter hypermethylation in breast cancer cells and restored by demethylation [PMID:32692756]. Beyond its tumor-suppressor role, SHISA3 forms a complex with HSPA8 in macrophages to activate NF-κB signaling and maintain M1 polarization, enhancing phagocytosis, antigen presentation, and CD8+ T cell-mediated antitumor immunity [PMID:39054639]. Transcriptionally, SHISA3 is controlled by KLF15-Wnt networks in the heart and induced by DAMP/PAMP-driven NF-κB signaling in immune cells [PMID:31582141, PMID:39054639].","teleology":[{"year":2014,"claim":"Established SHISA3 as a functional tumor suppressor by linking it mechanistically to Wnt/β-catenin signaling, answering whether its anti-tumor effect had a defined pathway readout.","evidence":"Microarray/pathway analysis, in vitro invasion assays, and orthotopic mouse tumor models with β-catenin degradation readout in lung cancer","pmids":["25036006"],"confidence":"Medium","gaps":["Did not resolve the molecular step at which SHISA3 promotes β-catenin degradation","No direct binding partner identified"]},{"year":2018,"claim":"Defined a metastasis-suppressing axis by placing SHISA3 upstream of TRIM21-SGSM1-MAPK, explaining how it blocks invasion independent of Wnt.","evidence":"Co-IP, ubiquitination assays, and SGSM1 knockdown rescue in nasopharyngeal carcinoma with in vitro and in vivo metastasis models","pmids":["30573520"],"confidence":"High","gaps":["Mechanism by which SHISA3 inhibits TRIM21-mediated ubiquitination of SGSM1 not structurally defined","Direct SHISA3-TRIM21 vs SHISA3-SGSM1 binding hierarchy not fully resolved"]},{"year":2019,"claim":"Clarified the cell-biological mechanism of Wnt antagonism by showing SHISA3 blocks Frizzled receptor maturation/trafficking, while KO mice revealed family redundancy.","evidence":"Adenoviral overexpression, β-catenin translocation and Axin2 assays in osteoblasts, plus Shisa3 KO mouse phenotyping","pmids":["31222549"],"confidence":"Medium","gaps":["No bone phenotype in KO leaves the in vivo role uncertain due to Shisa family redundancy","Direct SHISA3-Frizzled interaction not biochemically demonstrated"]},{"year":2019,"claim":"Extended SHISA3 function to RTK signaling by identifying FGFR1/3 interaction that suppresses AKT/mTOR and cancer stem cell traits, connecting it to drug resistance.","evidence":"Co-IP, Western blot of AKT/mTOR, sphere formation, and in vivo NOD-SCID models in lung adenocarcinoma","pmids":["31801598"],"confidence":"Medium","gaps":["Whether FGFR binding and Wnt antagonism are independent or coupled is unresolved","Single-lab interaction without reciprocal structural validation"]},{"year":2019,"claim":"Placed SHISA3 within a tissue transcriptional network by showing KLF15-Wnt control of its expression in vascular cells of fetal and remodeling hearts.","evidence":"Transcriptomics, Klf15 KO mice, Wnt-modulated and pressure-overload/ischemia models, and human KLF15 KO stem cell-derived myocardium","pmids":["31582141"],"confidence":"Medium","gaps":["Functional consequence of SHISA3 expression in cardiac vascular cells not determined","Whether cardiac SHISA3 acts through the same Frizzled/β-catenin mechanism not tested"]},{"year":2024,"claim":"Revealed a non-cancer-cell role: SHISA3 partners with HSPA8 to activate NF-κB and sustain M1 macrophage polarization, linking it to antitumor immunity.","evidence":"mRNA delivery, SHISA3-HSPA8 co-IP, NF-κB and phagocytosis/antigen presentation assays, and Shisa3 KO combination immunotherapy in vivo","pmids":["39054639"],"confidence":"Medium","gaps":["Molecular mechanism by which the SHISA3-HSPA8 complex activates NF-κB is undefined","Reconciliation of NF-κB-activating immune role with Wnt-antagonist tumor-cell role not addressed"]},{"year":null,"claim":"How SHISA3's distinct activities — Frizzled trafficking blockade, FGFR binding, TRIM21-SGSM1 regulation, and HSPA8-NF-κB complex formation — are integrated within a single protein and across cell types remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model defining the domains responsible for each interaction","No unifying explanation for context-dependent (tumor-suppressive vs immune-activating) functions"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[1,3]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,2,3]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[5]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[0,1]}],"complexes":[],"partners":["TRIM21","SGSM1","FGFR1","FGFR3","HSPA8"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"A0PJX4","full_name":"Protein shisa-3 homolog","aliases":[],"length_aa":238,"mass_kda":25.8,"function":"Plays an essential role in the maturation of presomitic mesoderm cells by individual attenuation of both FGF and WNT signaling","subcellular_location":"Endoplasmic reticulum membrane","url":"https://www.uniprot.org/uniprotkb/A0PJX4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/SHISA3","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/SHISA3","total_profiled":1310},"omim":[{"mim_id":"617325","title":"SHISA FAMILY, MEMBER 3; SHISA3","url":"https://www.omim.org/entry/617325"},{"mim_id":"617324","title":"SHISA FAMILY, MEMBER 2; SHISA2","url":"https://www.omim.org/entry/617324"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Lipid droplets","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"heart muscle","ntpm":13.5},{"tissue":"kidney","ntpm":19.3}],"url":"https://www.proteinatlas.org/search/SHISA3"},"hgnc":{"alias_symbol":["hShisa3"],"prev_symbol":[]},"alphafold":{"accession":"A0PJX4","domains":[{"cath_id":"-","chopping":"25-91","consensus_level":"medium","plddt":88.5928,"start":25,"end":91},{"cath_id":"1.20.5","chopping":"94-125","consensus_level":"medium","plddt":89.4134,"start":94,"end":125}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/A0PJX4","model_url":"https://alphafold.ebi.ac.uk/files/AF-A0PJX4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-A0PJX4-F1-predicted_aligned_error_v6.png","plddt_mean":67.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=SHISA3","jax_strain_url":"https://www.jax.org/strain/search?query=SHISA3"},"sequence":{"accession":"A0PJX4","fasta_url":"https://rest.uniprot.org/uniprotkb/A0PJX4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/A0PJX4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/A0PJX4"}},"corpus_meta":[{"pmid":"30573520","id":"PMC_30573520","title":"Hypermethylation of SHISA3 Promotes Nasopharyngeal Carcinoma Metastasis by Reducing SGSM1 Stability.","date":"2018","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/30573520","citation_count":40,"is_preprint":false},{"pmid":"31801598","id":"PMC_31801598","title":"Shisa3 brakes resistance to EGFR-TKIs in lung adenocarcinoma by suppressing cancer stem cell properties.","date":"2019","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/31801598","citation_count":38,"is_preprint":false},{"pmid":"25036006","id":"PMC_25036006","title":"Shisa3 is associated with prolonged survival through promoting β-catenin degradation in lung cancer.","date":"2014","source":"American journal of respiratory and critical care medicine","url":"https://pubmed.ncbi.nlm.nih.gov/25036006","citation_count":35,"is_preprint":false},{"pmid":"39054639","id":"PMC_39054639","title":"SHISA3 Reprograms Tumor-Associated Macrophages Toward an Antitumoral Phenotype and Enhances Cancer Immunotherapy.","date":"2024","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/39054639","citation_count":29,"is_preprint":false},{"pmid":"31582141","id":"PMC_31582141","title":"KLF15-Wnt-Dependent Cardiac Reprogramming Up-Regulates SHISA3 in the Mammalian Heart.","date":"2019","source":"Journal of the American College of Cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/31582141","citation_count":25,"is_preprint":false},{"pmid":"32692756","id":"PMC_32692756","title":"SHISA3, an antagonist of the Wnt/β-catenin signaling, is epigenetically silenced and its ectopic expression suppresses growth in breast cancer.","date":"2020","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/32692756","citation_count":12,"is_preprint":false},{"pmid":"31222549","id":"PMC_31222549","title":"The Shisa3 knockout mouse exhibits normal bone phenotype.","date":"2019","source":"Journal of bone and mineral metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/31222549","citation_count":5,"is_preprint":false},{"pmid":"36686193","id":"PMC_36686193","title":"Polymorphisms in SHISA3 and RFC3 genes and their association with feed conversion ratio in Hu sheep.","date":"2023","source":"Frontiers in veterinary science","url":"https://pubmed.ncbi.nlm.nih.gov/36686193","citation_count":4,"is_preprint":false},{"pmid":"28639485","id":"PMC_28639485","title":"Increased SHISA3 expression characterizes chronic lymphocytic leukemia patients sensitive to lenalidomide.","date":"2017","source":"Leukemia & lymphoma","url":"https://pubmed.ncbi.nlm.nih.gov/28639485","citation_count":4,"is_preprint":false},{"pmid":"33642344","id":"PMC_33642344","title":"Hypermethylation of SHISA3 DNA as a blood-based biomarker for colorectal cancer.","date":"2021","source":"The Chinese journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/33642344","citation_count":2,"is_preprint":false},{"pmid":"37433260","id":"PMC_37433260","title":"Combining a tetracycline (Tet)-inducible gRNA system and CRISPRa for titratable and timely controlled enhancement of endogenous SHISA3 activation in human induced pluripotent stem cells (hiPSC).","date":"2023","source":"Stem cell research","url":"https://pubmed.ncbi.nlm.nih.gov/37433260","citation_count":2,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.08.18.670897","title":"Single Cell RNA sequencing reveals transitional states and signaling shifts in nephron progenitor cells of the late-gestation rhesus macaque kidney","date":"2025-08-22","source":"bioRxiv","url":"https://doi.org/10.1101/2025.08.18.670897","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":8803,"output_tokens":2064,"usd":0.028685,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9080,"output_tokens":2680,"usd":0.0562,"stage2_stop_reason":"end_turn"},"total_usd":0.084885,"stage1_batch_id":"msgbatch_013LWZZT6MWaqB9qyTSeQBmU","stage2_batch_id":"msgbatch_0194ZzQaTGNpruoNJBbEMbt3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2018,\n      \"finding\": \"SHISA3 suppresses NPC cell invasion and metastasis by impeding TRIM21-mediated ubiquitination and degradation of SGSM1, thereby inhibiting MAPK pathway activation. Silencing SGSM1 abrogated the inhibitory effect of SHISA3 on cell migration and invasion, placing SHISA3 upstream of TRIM21-SGSM1-MAPK axis.\",\n      \"method\": \"In vitro invasion/migration assays, in vivo metastasis models, co-immunoprecipitation, ubiquitination assays, SGSM1 knockdown rescue experiments\",\n      \"journal\": \"Cancer Research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal functional epistasis (SGSM1 KD rescue), IP-based interaction, in vitro and in vivo phenotypes, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"30573520\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"SHISA3 accelerates β-catenin degradation to suppress WNT signaling, thereby inhibiting tumorigenesis and invasion in lung cancer cells in vitro and reducing tumor growth and metastasis in mouse models.\",\n      \"method\": \"Microarray/pathway analysis, in vitro invasion/migration assays, subcutaneous and orthotopic mouse tumor models, β-catenin degradation assays\",\n      \"journal\": \"American Journal of Respiratory and Critical Care Medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro and in vivo functional assays with mechanistic readout (β-catenin degradation), single lab, multiple methods\",\n      \"pmids\": [\"25036006\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"SHISA3 interacts with FGFR1 and FGFR3 to inhibit AKT/mTOR signaling, thereby suppressing cancer stem cell properties and reversing EGFR-TKI resistance in lung adenocarcinoma.\",\n      \"method\": \"Immunoprecipitation, Western blot, sphere formation assay, Transwell assay, CCK8, in vivo NOD-SCID mouse model, pharmacological inhibition\",\n      \"journal\": \"Journal of Experimental & Clinical Cancer Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-immunoprecipitation demonstrating FGFR1/3 interaction, downstream signaling (AKT/mTOR) by Western blot, in vivo validation, single lab\",\n      \"pmids\": [\"31801598\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"SHISA3 blocks maturation and transportation of Frizzled receptors to the cell surface, inhibiting Wnt/β-catenin signaling; adenovirus-mediated overexpression of Shisa3 significantly inhibited Wnt3a-induced nuclear translocation of β-catenin and mRNA expression of Wnt target gene Axin2 in osteoblasts. Shisa3 knockout mice showed no bone phenotype, suggesting functional redundancy with other Shisa family members.\",\n      \"method\": \"Adenovirus-mediated gene transfer, β-catenin nuclear translocation assay, Axin2 mRNA expression, micro-CT, Shisa3 KO mouse phenotyping\",\n      \"journal\": \"Journal of Bone and Mineral Metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct functional assay (Wnt3a-induced β-catenin translocation inhibition) plus KO mouse phenotyping, single lab, multiple methods\",\n      \"pmids\": [\"31222549\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"SHISA3 expression in the heart is regulated by KLF15-Wnt-dependent transcriptional networks; KLF15 represses Wnt-dependent transcriptional signaling postnatally, and loss of KLF15 leads to Wnt-dependent SHISA3 upregulation. SHISA3 is primarily expressed in vascular cells (VCs) in fetal hearts and during pathological remodeling, conserved in mouse and human models.\",\n      \"method\": \"Transcriptomic bioinformatics, Klf15 KO mouse phenotyping, Wnt-signaling modulated hearts, pressure overload and myocardial ischemia models, human KLF15 KO embryonic stem cells, engineered human myocardium\",\n      \"journal\": \"Journal of the American College of Cardiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO models plus human stem cell validation, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"31582141\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"SHISA3 expression in macrophages is induced by DAMPs/PAMPs via NF-κB transcription factors. Reciprocally, SHISA3 forms a complex with HSPA8 to activate NF-κB signaling, maintaining M1 polarization of macrophages, increasing phagocytosis and antigen presentation, and promoting CD8+ T cell-mediated antitumor immunity. Shisa3 knockout abolished the antitumor efficacy of TLR4 agonist MPLA combined with PD-1 antibody.\",\n      \"method\": \"mRNA delivery, co-immunoprecipitation (SHISA3-HSPA8 complex), NF-κB signaling assays, phagocytosis and antigen presentation assays, Shisa3 KO mouse experiments, combination immunotherapy in vivo\",\n      \"journal\": \"Advanced Science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-immunoprecipitation of SHISA3-HSPA8, NF-κB pathway validation, KO functional rescue in vivo, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"39054639\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Ectopic expression of SHISA3 in breast cancer cell lines (MDA-MB231, MCF-7) significantly reduced proliferation and migration. SHISA3 acts as an antagonist of Wnt/β-catenin signaling and its promoter is epigenetically silenced by hypermethylation; 5-aza-2'-deoxycytidine treatment restored SHISA3 expression.\",\n      \"method\": \"Ectopic expression, proliferation and migration assays, methylation-specific PCR, 5-aza-2'-deoxycytidine demethylation treatment\",\n      \"journal\": \"PLoS One\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — functional assays (proliferation/migration) with ectopic expression, demethylation rescue, single lab, multiple methods but no direct mechanistic protein interaction shown\",\n      \"pmids\": [\"32692756\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"SHISA3 is a tumor suppressor and Wnt/β-catenin antagonist that blocks Frizzled receptor maturation/transport to inhibit Wnt signaling and accelerate β-catenin degradation; it also suppresses invasion and metastasis via a TRIM21-SGSM1-MAPK axis, interacts with FGFR1/3 to inhibit AKT/mTOR signaling and cancer stem cell properties, and in macrophages forms a complex with HSPA8 to activate NF-κB signaling and maintain M1 polarization, with its expression regulated by KLF15-Wnt transcriptional networks in the heart and by DAMPs/PAMPs-induced NF-κB signaling in immune cells.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"SHISA3 is a tumor-suppressive antagonist of Wnt/\\u03b2-catenin signaling that also engages receptor tyrosine kinase and innate immune pathways to restrain tumor growth, invasion, and metastasis [#1, #3]. It inhibits Wnt signaling by blocking maturation and transport of Frizzled receptors to the cell surface, preventing Wnt3a-induced nuclear translocation of \\u03b2-catenin and expression of the target gene Axin2, and by accelerating \\u03b2-catenin degradation [#1, #3]. In carcinoma cells SHISA3 suppresses invasion and metastasis by impeding TRIM21-mediated ubiquitination and degradation of SGSM1, thereby dampening MAPK pathway activation [#0], and it interacts with FGFR1 and FGFR3 to inhibit AKT/mTOR signaling, suppress cancer stem cell properties, and reverse EGFR-TKI resistance [#2]. Its expression is silenced by promoter hypermethylation in breast cancer cells and restored by demethylation [#6]. Beyond its tumor-suppressor role, SHISA3 forms a complex with HSPA8 in macrophages to activate NF-\\u03baB signaling and maintain M1 polarization, enhancing phagocytosis, antigen presentation, and CD8+ T cell-mediated antitumor immunity [#5]. Transcriptionally, SHISA3 is controlled by KLF15-Wnt networks in the heart and induced by DAMP/PAMP-driven NF-\\u03baB signaling in immune cells [#4, #5].\",\n  \"teleology\": [\n    {\n      \"year\": 2014,\n      \"claim\": \"Established SHISA3 as a functional tumor suppressor by linking it mechanistically to Wnt/\\u03b2-catenin signaling, answering whether its anti-tumor effect had a defined pathway readout.\",\n      \"evidence\": \"Microarray/pathway analysis, in vitro invasion assays, and orthotopic mouse tumor models with \\u03b2-catenin degradation readout in lung cancer\",\n      \"pmids\": [\"25036006\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not resolve the molecular step at which SHISA3 promotes \\u03b2-catenin degradation\", \"No direct binding partner identified\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined a metastasis-suppressing axis by placing SHISA3 upstream of TRIM21-SGSM1-MAPK, explaining how it blocks invasion independent of Wnt.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, and SGSM1 knockdown rescue in nasopharyngeal carcinoma with in vitro and in vivo metastasis models\",\n      \"pmids\": [\"30573520\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which SHISA3 inhibits TRIM21-mediated ubiquitination of SGSM1 not structurally defined\", \"Direct SHISA3-TRIM21 vs SHISA3-SGSM1 binding hierarchy not fully resolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Clarified the cell-biological mechanism of Wnt antagonism by showing SHISA3 blocks Frizzled receptor maturation/trafficking, while KO mice revealed family redundancy.\",\n      \"evidence\": \"Adenoviral overexpression, \\u03b2-catenin translocation and Axin2 assays in osteoblasts, plus Shisa3 KO mouse phenotyping\",\n      \"pmids\": [\"31222549\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No bone phenotype in KO leaves the in vivo role uncertain due to Shisa family redundancy\", \"Direct SHISA3-Frizzled interaction not biochemically demonstrated\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended SHISA3 function to RTK signaling by identifying FGFR1/3 interaction that suppresses AKT/mTOR and cancer stem cell traits, connecting it to drug resistance.\",\n      \"evidence\": \"Co-IP, Western blot of AKT/mTOR, sphere formation, and in vivo NOD-SCID models in lung adenocarcinoma\",\n      \"pmids\": [\"31801598\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether FGFR binding and Wnt antagonism are independent or coupled is unresolved\", \"Single-lab interaction without reciprocal structural validation\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Placed SHISA3 within a tissue transcriptional network by showing KLF15-Wnt control of its expression in vascular cells of fetal and remodeling hearts.\",\n      \"evidence\": \"Transcriptomics, Klf15 KO mice, Wnt-modulated and pressure-overload/ischemia models, and human KLF15 KO stem cell-derived myocardium\",\n      \"pmids\": [\"31582141\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of SHISA3 expression in cardiac vascular cells not determined\", \"Whether cardiac SHISA3 acts through the same Frizzled/\\u03b2-catenin mechanism not tested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed a non-cancer-cell role: SHISA3 partners with HSPA8 to activate NF-\\u03baB and sustain M1 macrophage polarization, linking it to antitumor immunity.\",\n      \"evidence\": \"mRNA delivery, SHISA3-HSPA8 co-IP, NF-\\u03baB and phagocytosis/antigen presentation assays, and Shisa3 KO combination immunotherapy in vivo\",\n      \"pmids\": [\"39054639\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism by which the SHISA3-HSPA8 complex activates NF-\\u03baB is undefined\", \"Reconciliation of NF-\\u03baB-activating immune role with Wnt-antagonist tumor-cell role not addressed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How SHISA3's distinct activities \\u2014 Frizzled trafficking blockade, FGFR binding, TRIM21-SGSM1 regulation, and HSPA8-NF-\\u03baB complex formation \\u2014 are integrated within a single protein and across cell types remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model defining the domains responsible for each interaction\", \"No unifying explanation for context-dependent (tumor-suppressive vs immune-activating) functions\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [1, 3]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 2, 3]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TRIM21\", \"SGSM1\", \"FGFR1\", \"FGFR3\", \"HSPA8\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}