{"gene":"SPINK7","run_date":"2026-06-10T07:46:40","timeline":{"discoveries":[{"year":2018,"finding":"SPINK7 depletion in esophageal epithelial cells increases urokinase plasminogen-type activator (uPA) activity, which promotes uPA receptor (uPAR)-dependent eosinophil activation, barrier dysfunction, and production of TSLP; treatment with α1-antitrypsin (broad-spectrum anti-serine protease) reversed pathologic features of SPINK7 silencing. Genetic epistasis was demonstrated between variants in TSLP and the uPA-encoding gene PLAU in vivo.","method":"siRNA silencing in esophageal epithelial progenitor cells and primary esophageal epithelial cells; uPA activity assays; α1-antitrypsin rescue experiments; genetic epistasis analysis","journal":"Science translational medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (KD with defined cellular phenotypes, enzymatic activity assay, pharmacological rescue, in vivo epistasis), replicated across cell types","pmids":["29875205"],"is_preprint":false},{"year":2003,"finding":"SPINK7/ECRG2 directly interacts with metallothionein 2A (MT2A), confirmed by GST pull-down in vitro and co-immunoprecipitation in vivo. ECRG2 co-localizes with MT2A predominantly in nuclei. ECRG2 inhibits cell proliferation and induces apoptosis in esophageal cancer cells; MT2A modulates this effect.","method":"Yeast two-hybrid screen, GST pull-down assay, co-immunoprecipitation, confocal microscopy, transfection/co-transfection with antisense constructs","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal binding confirmed by pulldown and Co-IP, localization by confocal, functional readout by proliferation/apoptosis assays; single lab","pmids":["12646258"],"is_preprint":false},{"year":2007,"finding":"SPINK7/ECRG2 binds directly to two species of uPA (55 kDa and 33 kDa) and reduces uPA/plasmin proteolytic activity (measured by D-Val-Phe-Lys-p-nitroanilide substrate cleavage), thereby inhibiting cancer cell migration, invasion, and in vivo metastasis.","method":"Binding affinity assays, co-immunoprecipitation from conditioned medium, plasmin substrate activity assay (absorbance at 405 nm), in vitro migration/invasion assays, tail-vein metastasis model in nude mice","journal":"Carcinogenesis","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — direct enzymatic activity assay combined with binding Co-IP and in vivo metastasis; replicated in subsequent studies","pmids":["17602171"],"is_preprint":false},{"year":2009,"finding":"SPINK7/ECRG2 binds specifically to the kringle domain of uPA, forms a complex with uPA·uPAR, modifies the dynamic association of uPAR with β1 integrins, and disrupts downstream Src/MAP kinase signaling to suppress cell migration and invasion. Depletion of ECRG2 enhances uPAR–β1 integrin association and elevates basal Src/MAP kinase activation.","method":"Co-immunoprecipitation, in vitro Matrigel migration/invasion assay, domain-mapping binding studies, siRNA depletion, western blotting for Src/MAPK pathway","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, domain-level binding, multiple cell lines, loss-of-function with defined signaling readout; replicated across cancer cell lines","pmids":["19717562"],"is_preprint":false},{"year":2009,"finding":"SPINK7/ECRG2 inhibits proteolytic activities of uPA/plasmin and MMP2, prevents uPAR cleavage, disrupts the association of soluble uPAR fragment (sD2D3) with FPRL1 receptor, and thereby impairs FPRL1-dependent cell migration and invasion. ECRG2 depletion elevates uPA/plasmin and MMP2 activities and enhances uPAR–FPRL1 association.","method":"Protease activity assays (uPA/plasmin, MMP2), co-immunoprecipitation, siRNA knockdown, in vitro invasion assay","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — enzymatic activity assays and Co-IP in multiple cell lines, single lab","pmids":["19796867"],"is_preprint":false},{"year":2007,"finding":"SPINK7/ECRG2 localizes to centrosomes during interphase and to kinetochores during mitosis. ECRG2 depletion destabilizes p53, downregulates p21, increases cyclin E/CDK2 activity leading to centrosome amplification, abolishes p53 localization to centrosomes, and reduces BUBR1 protein levels causing spindle assembly checkpoint failure. Overexpression of ECRG2 restores p53-dependent suppression of centrosome duplication.","method":"Immunofluorescence localization, siRNA knockdown, western blotting (p53, p21, cyclin E, CDK2, BUBR1), CDK2 kinase activity assay, centrosome counting, spindle phenotype analysis, overexpression rescue","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiments with functional consequence, loss- and gain-of-function with defined molecular readouts; single lab","pmids":["18162463"],"is_preprint":false},{"year":2015,"finding":"SPINK7/ECRG2 negatively regulates the RNA-binding protein HuR by promoting HuR ubiquitination and degradation, thereby destabilizing XIAP mRNA and reducing XIAP protein levels, activating caspases, and inducing apoptosis in cancer cells. A cancer-derived V30E mutant of ECRG2 fails to suppress HuR or XIAP and confers resistance to anticancer drugs.","method":"Co-immunoprecipitation, ubiquitination assay, mRNA stability assay, western blotting, caspase activation assay, mutant ECRG2 (V30E) functional studies, drug resistance assay","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (ubiquitination assay, mRNA stability, caspase activation, natural mutant validation) in a single rigorous study","pmids":["26434587"],"is_preprint":false},{"year":2020,"finding":"SPINK7/ECRG2 is a direct transcriptional target of p53: two p53-binding sites were identified in the ECRG2 promoter, DNA damage enhances p53 binding to these sites and induces ECRG2 promoter activity in a p53-dependent manner. A natural promoter variant (small deletion present in ~38.5% of world population) is defective in responding to p53 and DNA damage. ECRG2 genetic disruption enhances cell survival after DNA-damaging drug treatments even when p53 is induced.","method":"Promoter-reporter (luciferase) assay, ChIP (chromatin immunoprecipitation), RT-PCR, western blotting, ECRG2 knockout cells with drug survival assays","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — ChIP confirming direct p53 binding, promoter-reporter assays, loss-of-function with functional readout; single lab but multiple orthogonal methods","pmids":["32681017"],"is_preprint":false},{"year":2021,"finding":"In a murine colitis model, Spink7 is significantly upregulated and is primarily expressed in neutrophils. Spink7-deficient mice showed enhanced colitis severity. Bone marrow reconstitution experiments established that immune-compartment expression of Spink7 is the main contributor to its protective role. Loss of Spink7 leads to augmented production of multiple chemokines and cytokines.","method":"Spink7 knockout mice, DSS-induced colitis model, bone marrow reconstitution, cytokine/chemokine measurement, qPCR, histopathology","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mice with defined colitis phenotype, bone marrow reconstitution establishing immune compartment contribution; single lab","pmids":["33722746"],"is_preprint":false},{"year":2025,"finding":"Spink7 is upregulated in differentiated epidermal granular keratinocytes during the proliferative phase of wound healing. Loss of Spink7 (siRNA knockdown or KO) delays wound closure with sustained neutrophil infiltration, elevated proteolytic activities of uPA, MMP2/9, and KLK5/7, augmented chemokine/cytokine production, and impaired M2 macrophage polarization. Inhibiting the KLK5/7 downstream effector PAR2 activation worsens the KO phenotype.","method":"siRNA knockdown in wounds, Spink7 KO mice, wound healing assays, protease activity measurement (uPA, MMP2/9, KLK5/7), cytokine/chemokine multiplex assay, immunofluorescence/IHC, microarray, PAR2 inhibition epistasis","journal":"Clinical and translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO and KD with defined molecular phenotypes, protease activity assays, epistasis with PAR2; single lab","pmids":["40147022"],"is_preprint":false},{"year":2026,"finding":"The transcription factor OVOL1 directly regulates SPINK7 promoter activity in esophageal epithelium. OVOL1 overexpression increases SPINK7 expression; OVOL1 depletion decreases SPINK7, impairs epithelial barrier, and increases TSLP production. IL-13 abrogates OVOL1 nuclear translocation and promotes OVOL1 protein degradation via the esophageal-specific cysteine protease calpain-14, thereby suppressing SPINK7 expression in eosinophilic esophagitis.","method":"OVOL1 overexpression and siRNA depletion, promoter-reporter assay, nuclear fractionation/immunofluorescence for OVOL1 localization, calpain-14 inhibition/depletion, barrier function assay, TSLP ELISA, human biopsy correlation","journal":"JCI insight","confidence":"High","confidence_rationale":"Tier 2 / Moderate — promoter-reporter, nuclear localization assays, protease-dependent degradation with rescue, functional barrier readout; multiple orthogonal methods in single rigorous study","pmids":["42048165"],"is_preprint":false}],"current_model":"SPINK7 (ECRG2) is a Kazal-type serine protease inhibitor that maintains esophageal epithelial barrier integrity and suppresses allergic and inflammatory responses by inhibiting uPA (and related proteases including MMP2/9, KLK5/7), thereby limiting uPAR/β1-integrin/Src/MAPK-driven cell migration and uPAR/FPRL1 signaling; its expression is transcriptionally activated by p53 in response to DNA damage and positively regulated by the transcription factor OVOL1, while IL-13-activated calpain-14 degrades OVOL1 to suppress SPINK7 in eosinophilic esophagitis; additionally, SPINK7 promotes cell cycle fidelity by stabilizing p53 at centrosomes/kinetochores and maintaining spindle checkpoint protein BUBR1, and induces apoptosis via negative regulation of HuR-mediated XIAP mRNA stability."},"narrative":{"mechanistic_narrative":"SPINK7 (ECRG2) is a Kazal-type serine protease inhibitor that safeguards epithelial barrier integrity and restrains inflammatory and tumor-invasive programs by neutralizing extracellular proteolytic activity [PMID:29875205, PMID:17602171]. Its core biochemical activity is direct binding to and inhibition of urokinase plasminogen activator (uPA): SPINK7 engages the kringle domain of uPA, forms a complex with uPA·uPAR, and suppresses uPA/plasmin and MMP2 proteolysis [PMID:17602171, PMID:19717562, PMID:19796867]. Through this inhibition it limits uPAR–β1 integrin association and downstream Src/MAP kinase signaling, and it blocks generation of the soluble uPAR fragment that engages FPRL1, thereby curbing cell migration and invasion and reducing in vivo metastasis [PMID:17602171, PMID:19717562, PMID:19796867]. In epithelia, loss of SPINK7 unleashes uPA activity, driving uPAR-dependent eosinophil activation, barrier dysfunction, and TSLP production, with broad serine-protease inhibition reversing these defects [PMID:29875205]; analogous protease-driven phenotypes underlie its protective roles in colitis and cutaneous wound healing, where SPINK7 loss elevates uPA, MMP2/9, and KLK5/7 activity and amplifies chemokine/cytokine output [PMID:33722746, PMID:40147022]. SPINK7 expression is transcriptionally controlled by p53 in response to DNA damage and by the epithelial transcription factor OVOL1, the latter being degraded by IL-13–activated calpain-14 to suppress SPINK7 in eosinophilic esophagitis [PMID:32681017, PMID:42048165]. Beyond protease inhibition, SPINK7 localizes to centrosomes and kinetochores to stabilize p53 and BUBR1 and preserve mitotic fidelity [PMID:18162463], and it promotes apoptosis by driving HuR ubiquitination to destabilize XIAP mRNA [PMID:26434587].","teleology":[{"year":2003,"claim":"Established the first molecular interaction and cellular activity for ECRG2, indicating a nuclear, growth-suppressive role in esophageal cancer cells.","evidence":"Yeast two-hybrid, GST pull-down, Co-IP, and confocal localization with proliferation/apoptosis assays in esophageal cancer cells","pmids":["12646258"],"confidence":"Medium","gaps":["Functional significance of the MT2A interaction not mechanistically resolved","Nuclear localization not reconciled with later secreted protease-inhibitor role","Single-lab finding"]},{"year":2007,"claim":"Defined SPINK7's biochemical activity as a direct uPA-binding inhibitor that suppresses proteolysis and metastasis, establishing it as a functional protease inhibitor.","evidence":"Binding assays, Co-IP from conditioned medium, plasmin substrate activity assay, in vitro invasion and tail-vein metastasis in nude mice","pmids":["17602171"],"confidence":"High","gaps":["Did not map the binding site on uPA","Did not connect inhibition to downstream signaling"]},{"year":2007,"claim":"Revealed a protease-independent role in mitotic fidelity, linking SPINK7 to p53 stabilization, centrosome integrity, and the spindle checkpoint.","evidence":"Immunofluorescence localization, siRNA knockdown, western blotting, CDK2 kinase assay, centrosome counting, and overexpression rescue","pmids":["18162463"],"confidence":"Medium","gaps":["Molecular mechanism by which SPINK7 stabilizes p53 and BUBR1 unknown","How a protease inhibitor localizes to centrosomes/kinetochores unresolved","Single lab"]},{"year":2009,"claim":"Mechanistically connected uPA inhibition to suppression of uPAR–β1 integrin/Src/MAPK signaling, explaining how SPINK7 limits cell migration.","evidence":"Domain-mapping binding, reciprocal Co-IP, siRNA depletion, Matrigel invasion, and Src/MAPK western blotting across cancer cell lines","pmids":["19717562"],"confidence":"High","gaps":["Stoichiometry of the SPINK7·uPA·uPAR complex not defined","In vivo relevance of the signaling axis not tested"]},{"year":2009,"claim":"Extended SPINK7's protease targets to MMP2 and showed it blocks uPAR cleavage and the sD2D3–FPRL1 migratory axis.","evidence":"uPA/plasmin and MMP2 activity assays, Co-IP, siRNA knockdown, and invasion assays in multiple cell lines","pmids":["19796867"],"confidence":"Medium","gaps":["Direct versus indirect inhibition of MMP2 not distinguished","Single lab"]},{"year":2015,"claim":"Identified an apoptosis-promoting mechanism via HuR ubiquitination and XIAP mRNA destabilization, and a cancer mutant that abolishes this activity.","evidence":"Co-IP, ubiquitination and mRNA-stability assays, caspase activation, and V30E mutant drug-resistance studies","pmids":["26434587"],"confidence":"High","gaps":["How SPINK7 drives HuR ubiquitination (E3 ligase involvement) unresolved","Relationship between this activity and protease inhibition unclear"]},{"year":2018,"claim":"Placed SPINK7 at the center of esophageal barrier homeostasis, showing its loss triggers uPA/uPAR-dependent eosinophil activation, barrier loss, and TSLP induction.","evidence":"siRNA silencing in esophageal epithelial cells, uPA activity assays, α1-antitrypsin rescue, and TSLP/PLAU genetic epistasis in vivo","pmids":["29875205"],"confidence":"High","gaps":["Upstream regulators of SPINK7 in EoE not yet identified at this stage","Direct protease responsible for barrier loss not fully isolated"]},{"year":2020,"claim":"Established SPINK7 as a direct DNA-damage-responsive p53 target gene, with a common defective promoter variant, linking it to genotoxic stress responses.","evidence":"Promoter-reporter luciferase assays, ChIP, RT-PCR, and knockout drug-survival assays","pmids":["32681017"],"confidence":"High","gaps":["Physiological consequence of the population promoter variant not tested in vivo","Connection between p53-driven expression and protease-inhibitor function unexplored"]},{"year":2021,"claim":"Demonstrated a protective immune-compartment role for SPINK7 in intestinal inflammation, expanding its function beyond epithelium to neutrophils.","evidence":"Spink7 KO mice, DSS colitis model, bone marrow reconstitution, and cytokine/chemokine profiling","pmids":["33722746"],"confidence":"Medium","gaps":["Protease target driving the neutrophil phenotype not defined","Single lab"]},{"year":2025,"claim":"Showed SPINK7 controls the resolution phase of wound healing by restraining a broad protease network (uPA, MMP2/9, KLK5/7) and shaping macrophage polarization.","evidence":"siRNA knockdown and KO mice in wound models, multi-protease activity assays, cytokine multiplex, and PAR2-inhibition epistasis","pmids":["40147022"],"confidence":"Medium","gaps":["Direct versus indirect inhibition of each protease not separated","Mechanism linking protease control to M2 polarization unresolved"]},{"year":2026,"claim":"Identified OVOL1 as a direct transcriptional activator of SPINK7 and a calpain-14/IL-13 axis that suppresses it, explaining SPINK7 loss in eosinophilic esophagitis.","evidence":"OVOL1 overexpression/depletion, promoter-reporter, nuclear localization assays, calpain-14 manipulation, barrier and TSLP readouts, and human biopsy correlation","pmids":["42048165"],"confidence":"High","gaps":["Whether OVOL1 and p53 act combinatorially on the promoter not addressed","In vivo reversal of EoE by restoring SPINK7 not demonstrated"]},{"year":null,"claim":"How SPINK7's extracellular protease-inhibitory function mechanistically integrates with its intracellular roles in p53/BUBR1 stabilization, HuR/XIAP-driven apoptosis, and centrosome/kinetochore localization remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model reconciling secreted protease inhibition with nuclear/mitotic activities","No structural model of SPINK7 protease complexes","E3 ligase and direct partners for intracellular functions unidentified"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,2,3,4]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[2,3]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[1]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[5]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[2]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[0,8,9]},{"term_id":"R-HSA-1474244","term_label":"Extracellular matrix organization","supporting_discovery_ids":[3,4]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[6]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[5]}],"complexes":[],"partners":["PLAU","PLAUR","ITGB1","MMP2","MT2A","ELAVL1","TP53","OVOL1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P58062","full_name":"Serine protease inhibitor Kazal-type 7","aliases":["Esophagus cancer-related gene 2 protein","ECRG-2"],"length_aa":85,"mass_kda":9.2,"function":"Probable serine protease inhibitor","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/P58062/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/SPINK7","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/SPINK7","total_profiled":1310},"omim":[{"mim_id":"617288","title":"SERINE PEPTIDASE INHIBITOR, KAZAL-TYPE, 7; SPINK7","url":"https://www.omim.org/entry/617288"},{"mim_id":"615868","title":"SERINE PEPTIDASE INHIBITOR, KAZAL-TYPE, 6; SPINK6","url":"https://www.omim.org/entry/615868"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"cervix","ntpm":517.3},{"tissue":"esophagus","ntpm":627.5},{"tissue":"vagina","ntpm":601.8}],"url":"https://www.proteinatlas.org/search/SPINK7"},"hgnc":{"alias_symbol":["ECG2","ECRG2"],"prev_symbol":[]},"alphafold":{"accession":"P58062","domains":[{"cath_id":"3.30.60.30","chopping":"26-81","consensus_level":"high","plddt":95.9239,"start":26,"end":81}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P58062","model_url":"https://alphafold.ebi.ac.uk/files/AF-P58062-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P58062-F1-predicted_aligned_error_v6.png","plddt_mean":89.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=SPINK7","jax_strain_url":"https://www.jax.org/strain/search?query=SPINK7"},"sequence":{"accession":"P58062","fasta_url":"https://rest.uniprot.org/uniprotkb/P58062.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P58062/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P58062"}},"corpus_meta":[{"pmid":"29875205","id":"PMC_29875205","title":"The antiprotease SPINK7 serves as an inhibitory checkpoint for esophageal epithelial inflammatory responses.","date":"2018","source":"Science translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29875205","citation_count":96,"is_preprint":false},{"pmid":"12646258","id":"PMC_12646258","title":"ECRG2, a novel candidate of tumor suppressor gene in the esophageal carcinoma, interacts directly with metallothionein 2A and links to apoptosis.","date":"2003","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/12646258","citation_count":57,"is_preprint":false},{"pmid":"22315007","id":"PMC_22315007","title":"MicroRNA-1322 regulates ECRG2 allele specifically and acts as a potential biomarker in patients with esophageal squamous cell carcinoma.","date":"2012","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/22315007","citation_count":45,"is_preprint":false},{"pmid":"26434587","id":"PMC_26434587","title":"Negative regulation of RNA-binding protein HuR by tumor-suppressor ECRG2.","date":"2015","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/26434587","citation_count":29,"is_preprint":false},{"pmid":"17602171","id":"PMC_17602171","title":"ECRG2 inhibits cancer cell migration, invasion and metastasis through the down-regulation of uPA/plasmin activity.","date":"2007","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/17602171","citation_count":29,"is_preprint":false},{"pmid":"24606472","id":"PMC_24606472","title":"Expression of bcl-2 and p53 in induction of esophageal cancer cell apoptosis by ECRG2 in combination with cisplatin.","date":"2014","source":"Asian Pacific journal of cancer prevention : APJCP","url":"https://pubmed.ncbi.nlm.nih.gov/24606472","citation_count":29,"is_preprint":false},{"pmid":"19717562","id":"PMC_19717562","title":"ECRG2 regulates cell migration/invasion through urokinase-type plasmin activator receptor (uPAR)/beta1 integrin pathway.","date":"2009","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19717562","citation_count":24,"is_preprint":false},{"pmid":"14639608","id":"PMC_14639608","title":"Short tandem repeat polymorphism in a novel esophageal cancer-related gene (ECRG2) implicates susceptibility to esophageal cancer in Chinese population.","date":"2004","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/14639608","citation_count":23,"is_preprint":false},{"pmid":"19796867","id":"PMC_19796867","title":"ECRG2 regulates ECM degradation and uPAR/FPRL1 pathway contributing cell invasion/migration.","date":"2009","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/19796867","citation_count":22,"is_preprint":false},{"pmid":"28348485","id":"PMC_28348485","title":"ECRG2 enhances the anti-cancer effects of cisplatin in cisplatin-resistant esophageal cancer cells via upregulation of p53 and downregulation of PCNA.","date":"2017","source":"World journal of gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/28348485","citation_count":21,"is_preprint":false},{"pmid":"18162463","id":"PMC_18162463","title":"ECRG2 disruption leads to centrosome amplification and spindle checkpoint defects contributing chromosome instability.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18162463","citation_count":20,"is_preprint":false},{"pmid":"34603317","id":"PMC_34603317","title":"SPINK7 Recognizes Fungi and Initiates Hemocyte-Mediated Immune Defense Against Fungal Infections.","date":"2021","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34603317","citation_count":15,"is_preprint":false},{"pmid":"33767253","id":"PMC_33767253","title":"SPINK7 expression changes accompanied by HER2, P53 and RB1 can be relevant in predicting oral squamous cell carcinoma at a molecular level.","date":"2021","source":"Scientific 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Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/33722746","citation_count":7,"is_preprint":false},{"pmid":"40147022","id":"PMC_40147022","title":"The antiprotease Spink7 promotes inflammation resolution by modulating multiple proteases activities during wound healing.","date":"2025","source":"Clinical and translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/40147022","citation_count":5,"is_preprint":false},{"pmid":"18618216","id":"PMC_18618216","title":"Association of ECRG2 TCA short tandem repeat polymorphism with the risk of oesophageal cancer in a North Indian population.","date":"2008","source":"Clinical and experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/18618216","citation_count":5,"is_preprint":false},{"pmid":"17352218","id":"PMC_17352218","title":"Short tandem repeat polymorphisms of exon 4 in Kazal-type gene ECRG2 in pancreatic carcinoma and chronic pancreatitis.","date":"2007","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/17352218","citation_count":4,"is_preprint":false},{"pmid":"25289046","id":"PMC_25289046","title":"Effect of ECRG2 in combination with cisplatin on the proliferation and apoptosis of EC9706 cells.","date":"2014","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/25289046","citation_count":4,"is_preprint":false},{"pmid":"38892042","id":"PMC_38892042","title":"ECRG2/SPINK7 Tumor Suppressor as Modulator of DNA Damage Response.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38892042","citation_count":2,"is_preprint":false},{"pmid":"42048165","id":"PMC_42048165","title":"IL-13 and calpain-14 suppress the expression of SPINK7 by regulating OVOL1 in eosinophilic esophagitis.","date":"2026","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/42048165","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14226,"output_tokens":3394,"usd":0.046794,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10860,"output_tokens":3864,"usd":0.07545,"stage2_stop_reason":"end_turn"},"total_usd":0.122244,"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\": \"SPINK7 depletion in esophageal epithelial cells increases urokinase plasminogen-type activator (uPA) activity, which promotes uPA receptor (uPAR)-dependent eosinophil activation, barrier dysfunction, and production of TSLP; treatment with α1-antitrypsin (broad-spectrum anti-serine protease) reversed pathologic features of SPINK7 silencing. Genetic epistasis was demonstrated between variants in TSLP and the uPA-encoding gene PLAU in vivo.\",\n      \"method\": \"siRNA silencing in esophageal epithelial progenitor cells and primary esophageal epithelial cells; uPA activity assays; α1-antitrypsin rescue experiments; genetic epistasis analysis\",\n      \"journal\": \"Science translational medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (KD with defined cellular phenotypes, enzymatic activity assay, pharmacological rescue, in vivo epistasis), replicated across cell types\",\n      \"pmids\": [\"29875205\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"SPINK7/ECRG2 directly interacts with metallothionein 2A (MT2A), confirmed by GST pull-down in vitro and co-immunoprecipitation in vivo. ECRG2 co-localizes with MT2A predominantly in nuclei. ECRG2 inhibits cell proliferation and induces apoptosis in esophageal cancer cells; MT2A modulates this effect.\",\n      \"method\": \"Yeast two-hybrid screen, GST pull-down assay, co-immunoprecipitation, confocal microscopy, transfection/co-transfection with antisense constructs\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal binding confirmed by pulldown and Co-IP, localization by confocal, functional readout by proliferation/apoptosis assays; single lab\",\n      \"pmids\": [\"12646258\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"SPINK7/ECRG2 binds directly to two species of uPA (55 kDa and 33 kDa) and reduces uPA/plasmin proteolytic activity (measured by D-Val-Phe-Lys-p-nitroanilide substrate cleavage), thereby inhibiting cancer cell migration, invasion, and in vivo metastasis.\",\n      \"method\": \"Binding affinity assays, co-immunoprecipitation from conditioned medium, plasmin substrate activity assay (absorbance at 405 nm), in vitro migration/invasion assays, tail-vein metastasis model in nude mice\",\n      \"journal\": \"Carcinogenesis\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — direct enzymatic activity assay combined with binding Co-IP and in vivo metastasis; replicated in subsequent studies\",\n      \"pmids\": [\"17602171\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"SPINK7/ECRG2 binds specifically to the kringle domain of uPA, forms a complex with uPA·uPAR, modifies the dynamic association of uPAR with β1 integrins, and disrupts downstream Src/MAP kinase signaling to suppress cell migration and invasion. Depletion of ECRG2 enhances uPAR–β1 integrin association and elevates basal Src/MAP kinase activation.\",\n      \"method\": \"Co-immunoprecipitation, in vitro Matrigel migration/invasion assay, domain-mapping binding studies, siRNA depletion, western blotting for Src/MAPK pathway\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, domain-level binding, multiple cell lines, loss-of-function with defined signaling readout; replicated across cancer cell lines\",\n      \"pmids\": [\"19717562\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"SPINK7/ECRG2 inhibits proteolytic activities of uPA/plasmin and MMP2, prevents uPAR cleavage, disrupts the association of soluble uPAR fragment (sD2D3) with FPRL1 receptor, and thereby impairs FPRL1-dependent cell migration and invasion. ECRG2 depletion elevates uPA/plasmin and MMP2 activities and enhances uPAR–FPRL1 association.\",\n      \"method\": \"Protease activity assays (uPA/plasmin, MMP2), co-immunoprecipitation, siRNA knockdown, in vitro invasion assay\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — enzymatic activity assays and Co-IP in multiple cell lines, single lab\",\n      \"pmids\": [\"19796867\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"SPINK7/ECRG2 localizes to centrosomes during interphase and to kinetochores during mitosis. ECRG2 depletion destabilizes p53, downregulates p21, increases cyclin E/CDK2 activity leading to centrosome amplification, abolishes p53 localization to centrosomes, and reduces BUBR1 protein levels causing spindle assembly checkpoint failure. Overexpression of ECRG2 restores p53-dependent suppression of centrosome duplication.\",\n      \"method\": \"Immunofluorescence localization, siRNA knockdown, western blotting (p53, p21, cyclin E, CDK2, BUBR1), CDK2 kinase activity assay, centrosome counting, spindle phenotype analysis, overexpression rescue\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiments with functional consequence, loss- and gain-of-function with defined molecular readouts; single lab\",\n      \"pmids\": [\"18162463\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"SPINK7/ECRG2 negatively regulates the RNA-binding protein HuR by promoting HuR ubiquitination and degradation, thereby destabilizing XIAP mRNA and reducing XIAP protein levels, activating caspases, and inducing apoptosis in cancer cells. A cancer-derived V30E mutant of ECRG2 fails to suppress HuR or XIAP and confers resistance to anticancer drugs.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, mRNA stability assay, western blotting, caspase activation assay, mutant ECRG2 (V30E) functional studies, drug resistance assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (ubiquitination assay, mRNA stability, caspase activation, natural mutant validation) in a single rigorous study\",\n      \"pmids\": [\"26434587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"SPINK7/ECRG2 is a direct transcriptional target of p53: two p53-binding sites were identified in the ECRG2 promoter, DNA damage enhances p53 binding to these sites and induces ECRG2 promoter activity in a p53-dependent manner. A natural promoter variant (small deletion present in ~38.5% of world population) is defective in responding to p53 and DNA damage. ECRG2 genetic disruption enhances cell survival after DNA-damaging drug treatments even when p53 is induced.\",\n      \"method\": \"Promoter-reporter (luciferase) assay, ChIP (chromatin immunoprecipitation), RT-PCR, western blotting, ECRG2 knockout cells with drug survival assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — ChIP confirming direct p53 binding, promoter-reporter assays, loss-of-function with functional readout; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"32681017\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In a murine colitis model, Spink7 is significantly upregulated and is primarily expressed in neutrophils. Spink7-deficient mice showed enhanced colitis severity. Bone marrow reconstitution experiments established that immune-compartment expression of Spink7 is the main contributor to its protective role. Loss of Spink7 leads to augmented production of multiple chemokines and cytokines.\",\n      \"method\": \"Spink7 knockout mice, DSS-induced colitis model, bone marrow reconstitution, cytokine/chemokine measurement, qPCR, histopathology\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mice with defined colitis phenotype, bone marrow reconstitution establishing immune compartment contribution; single lab\",\n      \"pmids\": [\"33722746\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Spink7 is upregulated in differentiated epidermal granular keratinocytes during the proliferative phase of wound healing. Loss of Spink7 (siRNA knockdown or KO) delays wound closure with sustained neutrophil infiltration, elevated proteolytic activities of uPA, MMP2/9, and KLK5/7, augmented chemokine/cytokine production, and impaired M2 macrophage polarization. Inhibiting the KLK5/7 downstream effector PAR2 activation worsens the KO phenotype.\",\n      \"method\": \"siRNA knockdown in wounds, Spink7 KO mice, wound healing assays, protease activity measurement (uPA, MMP2/9, KLK5/7), cytokine/chemokine multiplex assay, immunofluorescence/IHC, microarray, PAR2 inhibition epistasis\",\n      \"journal\": \"Clinical and translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO and KD with defined molecular phenotypes, protease activity assays, epistasis with PAR2; single lab\",\n      \"pmids\": [\"40147022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"The transcription factor OVOL1 directly regulates SPINK7 promoter activity in esophageal epithelium. OVOL1 overexpression increases SPINK7 expression; OVOL1 depletion decreases SPINK7, impairs epithelial barrier, and increases TSLP production. IL-13 abrogates OVOL1 nuclear translocation and promotes OVOL1 protein degradation via the esophageal-specific cysteine protease calpain-14, thereby suppressing SPINK7 expression in eosinophilic esophagitis.\",\n      \"method\": \"OVOL1 overexpression and siRNA depletion, promoter-reporter assay, nuclear fractionation/immunofluorescence for OVOL1 localization, calpain-14 inhibition/depletion, barrier function assay, TSLP ELISA, human biopsy correlation\",\n      \"journal\": \"JCI insight\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — promoter-reporter, nuclear localization assays, protease-dependent degradation with rescue, functional barrier readout; multiple orthogonal methods in single rigorous study\",\n      \"pmids\": [\"42048165\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"SPINK7 (ECRG2) is a Kazal-type serine protease inhibitor that maintains esophageal epithelial barrier integrity and suppresses allergic and inflammatory responses by inhibiting uPA (and related proteases including MMP2/9, KLK5/7), thereby limiting uPAR/β1-integrin/Src/MAPK-driven cell migration and uPAR/FPRL1 signaling; its expression is transcriptionally activated by p53 in response to DNA damage and positively regulated by the transcription factor OVOL1, while IL-13-activated calpain-14 degrades OVOL1 to suppress SPINK7 in eosinophilic esophagitis; additionally, SPINK7 promotes cell cycle fidelity by stabilizing p53 at centrosomes/kinetochores and maintaining spindle checkpoint protein BUBR1, and induces apoptosis via negative regulation of HuR-mediated XIAP mRNA stability.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"SPINK7 (ECRG2) is a Kazal-type serine protease inhibitor that safeguards epithelial barrier integrity and restrains inflammatory and tumor-invasive programs by neutralizing extracellular proteolytic activity [#0, #2]. Its core biochemical activity is direct binding to and inhibition of urokinase plasminogen activator (uPA): SPINK7 engages the kringle domain of uPA, forms a complex with uPA·uPAR, and suppresses uPA/plasmin and MMP2 proteolysis [#2, #3, #4]. Through this inhibition it limits uPAR–\\u03b21 integrin association and downstream Src/MAP kinase signaling, and it blocks generation of the soluble uPAR fragment that engages FPRL1, thereby curbing cell migration and invasion and reducing in vivo metastasis [#2, #3, #4]. In epithelia, loss of SPINK7 unleashes uPA activity, driving uPAR-dependent eosinophil activation, barrier dysfunction, and TSLP production, with broad serine-protease inhibition reversing these defects [#0]; analogous protease-driven phenotypes underlie its protective roles in colitis and cutaneous wound healing, where SPINK7 loss elevates uPA, MMP2/9, and KLK5/7 activity and amplifies chemokine/cytokine output [#8, #9]. SPINK7 expression is transcriptionally controlled by p53 in response to DNA damage and by the epithelial transcription factor OVOL1, the latter being degraded by IL-13\\u2013activated calpain-14 to suppress SPINK7 in eosinophilic esophagitis [#7, #10]. Beyond protease inhibition, SPINK7 localizes to centrosomes and kinetochores to stabilize p53 and BUBR1 and preserve mitotic fidelity [#5], and it promotes apoptosis by driving HuR ubiquitination to destabilize XIAP mRNA [#6].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established the first molecular interaction and cellular activity for ECRG2, indicating a nuclear, growth-suppressive role in esophageal cancer cells.\",\n      \"evidence\": \"Yeast two-hybrid, GST pull-down, Co-IP, and confocal localization with proliferation/apoptosis assays in esophageal cancer cells\",\n      \"pmids\": [\"12646258\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional significance of the MT2A interaction not mechanistically resolved\", \"Nuclear localization not reconciled with later secreted protease-inhibitor role\", \"Single-lab finding\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Defined SPINK7's biochemical activity as a direct uPA-binding inhibitor that suppresses proteolysis and metastasis, establishing it as a functional protease inhibitor.\",\n      \"evidence\": \"Binding assays, Co-IP from conditioned medium, plasmin substrate activity assay, in vitro invasion and tail-vein metastasis in nude mice\",\n      \"pmids\": [\"17602171\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not map the binding site on uPA\", \"Did not connect inhibition to downstream signaling\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Revealed a protease-independent role in mitotic fidelity, linking SPINK7 to p53 stabilization, centrosome integrity, and the spindle checkpoint.\",\n      \"evidence\": \"Immunofluorescence localization, siRNA knockdown, western blotting, CDK2 kinase assay, centrosome counting, and overexpression rescue\",\n      \"pmids\": [\"18162463\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism by which SPINK7 stabilizes p53 and BUBR1 unknown\", \"How a protease inhibitor localizes to centrosomes/kinetochores unresolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Mechanistically connected uPA inhibition to suppression of uPAR–\\u03b21 integrin/Src/MAPK signaling, explaining how SPINK7 limits cell migration.\",\n      \"evidence\": \"Domain-mapping binding, reciprocal Co-IP, siRNA depletion, Matrigel invasion, and Src/MAPK western blotting across cancer cell lines\",\n      \"pmids\": [\"19717562\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry of the SPINK7·uPA·uPAR complex not defined\", \"In vivo relevance of the signaling axis not tested\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Extended SPINK7's protease targets to MMP2 and showed it blocks uPAR cleavage and the sD2D3–FPRL1 migratory axis.\",\n      \"evidence\": \"uPA/plasmin and MMP2 activity assays, Co-IP, siRNA knockdown, and invasion assays in multiple cell lines\",\n      \"pmids\": [\"19796867\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct versus indirect inhibition of MMP2 not distinguished\", \"Single lab\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified an apoptosis-promoting mechanism via HuR ubiquitination and XIAP mRNA destabilization, and a cancer mutant that abolishes this activity.\",\n      \"evidence\": \"Co-IP, ubiquitination and mRNA-stability assays, caspase activation, and V30E mutant drug-resistance studies\",\n      \"pmids\": [\"26434587\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How SPINK7 drives HuR ubiquitination (E3 ligase involvement) unresolved\", \"Relationship between this activity and protease inhibition unclear\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Placed SPINK7 at the center of esophageal barrier homeostasis, showing its loss triggers uPA/uPAR-dependent eosinophil activation, barrier loss, and TSLP induction.\",\n      \"evidence\": \"siRNA silencing in esophageal epithelial cells, uPA activity assays, \\u03b11-antitrypsin rescue, and TSLP/PLAU genetic epistasis in vivo\",\n      \"pmids\": [\"29875205\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream regulators of SPINK7 in EoE not yet identified at this stage\", \"Direct protease responsible for barrier loss not fully isolated\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established SPINK7 as a direct DNA-damage-responsive p53 target gene, with a common defective promoter variant, linking it to genotoxic stress responses.\",\n      \"evidence\": \"Promoter-reporter luciferase assays, ChIP, RT-PCR, and knockout drug-survival assays\",\n      \"pmids\": [\"32681017\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological consequence of the population promoter variant not tested in vivo\", \"Connection between p53-driven expression and protease-inhibitor function unexplored\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated a protective immune-compartment role for SPINK7 in intestinal inflammation, expanding its function beyond epithelium to neutrophils.\",\n      \"evidence\": \"Spink7 KO mice, DSS colitis model, bone marrow reconstitution, and cytokine/chemokine profiling\",\n      \"pmids\": [\"33722746\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Protease target driving the neutrophil phenotype not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showed SPINK7 controls the resolution phase of wound healing by restraining a broad protease network (uPA, MMP2/9, KLK5/7) and shaping macrophage polarization.\",\n      \"evidence\": \"siRNA knockdown and KO mice in wound models, multi-protease activity assays, cytokine multiplex, and PAR2-inhibition epistasis\",\n      \"pmids\": [\"40147022\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct versus indirect inhibition of each protease not separated\", \"Mechanism linking protease control to M2 polarization unresolved\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identified OVOL1 as a direct transcriptional activator of SPINK7 and a calpain-14/IL-13 axis that suppresses it, explaining SPINK7 loss in eosinophilic esophagitis.\",\n      \"evidence\": \"OVOL1 overexpression/depletion, promoter-reporter, nuclear localization assays, calpain-14 manipulation, barrier and TSLP readouts, and human biopsy correlation\",\n      \"pmids\": [\"42048165\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether OVOL1 and p53 act combinatorially on the promoter not addressed\", \"In vivo reversal of EoE by restoring SPINK7 not demonstrated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How SPINK7's extracellular protease-inhibitory function mechanistically integrates with its intracellular roles in p53/BUBR1 stabilization, HuR/XIAP-driven apoptosis, and centrosome/kinetochore localization remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model reconciling secreted protease inhibition with nuclear/mitotic activities\", \"No structural model of SPINK7 protease complexes\", \"E3 ligase and direct partners for intracellular functions unidentified\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 2, 3, 4]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0, 8, 9]},\n      {\"term_id\": \"R-HSA-1474244\", \"supporting_discovery_ids\": [3, 4]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"PLAU\", \"PLAUR\", \"ITGB1\", \"MMP2\", \"MT2A\", \"ELAVL1\", \"TP53\", \"OVOL1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":6,"faith_total":6,"faith_pct":100.0}}