{"gene":"ECHDC2","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2013,"finding":"ECHDC2 is a mitochondrial protein expressed in cardiomyocytes but not cardiofibroblasts. Overexpression of ECHDC2 increased susceptibility to ischemia/reperfusion (I/R) injury, while knockdown enhanced resistance to I/R injury. ECHDC2 overexpression increased cellular levels of branched-chain amino acids leucine and valine (measured by GC-MS), but did not alter mitochondrial O2 consumption, metabolic intermediates, or ATP production.","method":"Subcellular fractionation/immunohistochemistry for localization; overexpression and knockdown with I/R injury assay; GC-MS metabolite analysis; Western blot; RT-PCR","journal":"Journal of the American Heart Association","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (localization, gain/loss-of-function, metabolomics) in a single lab; no independent replication","pmids":["24108764"],"is_preprint":false},{"year":2024,"finding":"ECHDC2 binds NEDD4 (E3 ubiquitin ligase) via co-immunoprecipitation, promotes ubiquitination and proteasomal degradation of MCCC2, and thereby suppresses the P38 MAPK pathway, leading to reduced aerobic glycolysis and inhibition of gastric cancer cell proliferation both in vitro and in vivo.","method":"Co-immunoprecipitation, Western blotting, immunofluorescence, colony formation, CCK8, EDU, glucose/lactate assays, subcutaneous xenograft in nude mice","journal":"Molecular medicine (Cambridge, Mass.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP demonstrating ECHDC2-NEDD4 interaction plus functional in vivo/in vitro validation; single lab, no independent replication","pmids":["38783226"],"is_preprint":false},{"year":2019,"finding":"ECHDC2 knockdown significantly inhibited suspension growth (anoikis escape) of high-grade serous ovarian cancer cells, implicating ECHDC2 as an effector of fatty acid metabolism required for survival in suspension.","method":"Genome-wide CRISPR/Cas9 knockout screen followed by targeted siRNA knockdown; suspension growth assay; non-targeted metabolomics","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — GeCKO screen plus targeted knockdown validation with functional readout; single study, metabolomic support","pmids":["31437751"],"is_preprint":false},{"year":2026,"finding":"ECHDC2 knockdown suppressed proliferation and migration of glioblastoma (GBM) cell lines, while overexpression had the opposite effect; enrichment analyses associated ECHDC2 activity with the PI3K/Akt signaling pathway in GBM.","method":"siRNA knockdown and overexpression in GBM cell lines; CCK-8, EdU, wound-healing, and Transwell migration assays; GSEA/KEGG pathway enrichment","journal":"Frontiers in genetics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, cellular phenotype established but pathway placement relies on bioinformatics enrichment without direct pathway validation","pmids":["41732157"],"is_preprint":false},{"year":2025,"finding":"Very high-dose vitamin D3 supplementation in pigs downregulated ECHDC2 protein levels in liver tissue, consistent with a role for ECHDC2 in mitochondrial β-oxidation pathways.","method":"Integrative transcriptomic and proteomic analysis of liver tissue; quantitative proteomics","journal":"The Journal of steroid biochemistry and molecular biology","confidence":"Low","confidence_rationale":"Tier 4 / Weak — protein-level change observed in a non-targeted proteomics study; no direct functional experiment on ECHDC2 itself","pmids":["40680899"],"is_preprint":false}],"current_model":"ECHDC2 is a mitochondrial protein involved in fatty acid β-oxidation and branched-chain amino acid metabolism; it promotes ubiquitination and degradation of MCCC2 via interaction with the E3 ligase NEDD4, thereby suppressing the P38 MAPK pathway and aerobic glycolysis, and its expression level modulates susceptibility to ischemia/reperfusion injury as well as proliferation and survival in multiple cancer contexts."},"narrative":{"mechanistic_narrative":"ECHDC2 is a mitochondrial protein linked to branched-chain amino acid and fatty acid metabolism that modulates cellular survival and proliferation across cardiac and cancer contexts [PMID:24108764, PMID:31437751]. In cardiomyocytes, ECHDC2 localizes to mitochondria, and its level sets susceptibility to ischemia/reperfusion injury—overexpression sensitizes cells while knockdown confers resistance—accompanied by elevated branched-chain amino acids leucine and valine without measurable changes in oxygen consumption or ATP production [PMID:24108764]. In gastric cancer, ECHDC2 binds the E3 ubiquitin ligase NEDD4 and promotes ubiquitination and proteasomal degradation of MCCC2, thereby suppressing P38 MAPK signaling, aerobic glycolysis, and tumor cell proliferation in vitro and in xenografts [PMID:38783226]. ECHDC2 is also required for anoikis escape and suspension growth of high-grade serous ovarian cancer cells, consistent with a role downstream of fatty acid metabolism [PMID:31437751]. Beyond these contexts and the NEDD4–MCCC2 axis, no enzymatic substrate or structural mechanism for ECHDC2 has been characterized in the available corpus.","teleology":[{"year":2013,"claim":"Established ECHDC2 as a mitochondrial protein whose abundance bidirectionally controls cardiomyocyte vulnerability to ischemia/reperfusion injury and influences branched-chain amino acid levels.","evidence":"Subcellular fractionation/IHC localization, gain- and loss-of-function with I/R injury assay, and GC-MS metabolomics in cardiomyocytes","pmids":["24108764"],"confidence":"Medium","gaps":["The enzymatic activity of ECHDC2 on BCAA or fatty acid intermediates was not directly demonstrated","Mechanism linking ECHDC2 to I/R susceptibility unresolved despite no change in O2 consumption or ATP","No in vivo cardiac model beyond cellular assays"]},{"year":2019,"claim":"Identified ECHDC2 as a fatty-acid-metabolism effector required for anoikis escape, extending its role from cardiac biology to cancer cell survival in suspension.","evidence":"Genome-wide CRISPR/Cas9 screen with siRNA validation, suspension growth assay, and non-targeted metabolomics in ovarian cancer cells","pmids":["31437751"],"confidence":"Medium","gaps":["The metabolic substrate of ECHDC2 driving suspension survival was not defined","No reciprocal rescue or overexpression to confirm specificity","Connection to a defined signaling pathway not established"]},{"year":2024,"claim":"Defined a molecular mechanism for ECHDC2's tumor-suppressive action: recruitment of NEDD4 to drive MCCC2 degradation, dampening P38 MAPK signaling and aerobic glycolysis.","evidence":"Reciprocal co-immunoprecipitation, ubiquitination and Western assays, glucose/lactate measurements, and subcutaneous xenografts in gastric cancer","pmids":["38783226"],"confidence":"Medium","gaps":["Whether ECHDC2 directly recruits NEDD4 or acts as an adaptor versus enzyme is not resolved","Single lab without independent replication","Reconciliation with pro-survival roles in ovarian/glioblastoma contexts not addressed"]},{"year":2026,"claim":"Showed that, in contrast to its gastric tumor-suppressive role, ECHDC2 promotes proliferation and migration of glioblastoma cells, associating it with PI3K/Akt signaling.","evidence":"siRNA knockdown and overexpression with proliferation/migration assays and GSEA/KEGG enrichment in GBM cell lines","pmids":["41732157"],"confidence":"Low","gaps":["PI3K/Akt placement rests on bioinformatic enrichment without direct pathway validation","Context-dependent opposing roles across cancers mechanistically unexplained","No in vivo confirmation"]},{"year":2025,"claim":"Provided correlative in vivo evidence linking ECHDC2 abundance to mitochondrial β-oxidation by showing high-dose vitamin D3 downregulates hepatic ECHDC2.","evidence":"Integrative transcriptomic and quantitative proteomic analysis of pig liver tissue","pmids":["40680899"],"confidence":"Low","gaps":["Protein-level change observed without any direct functional test of ECHDC2","Causal relationship between vitamin D3 and ECHDC2 not established","β-oxidation role inferred, not measured"]},{"year":null,"claim":"The direct biochemical activity of ECHDC2 and how it produces opposing proliferative outcomes across tissues remain unknown.","evidence":"No reconstituted enzymatic assay or structural characterization in the available corpus","pmids":[],"confidence":"Low","gaps":["No defined catalytic substrate or reaction for ECHDC2","Mechanism reconciling tumor-suppressor (gastric) versus pro-tumor (GBM) roles unresolved","No structural model or domain-level mechanism of NEDD4/MCCC2 engagement"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1]}],"localization":[{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[0,2]}],"complexes":[],"partners":["NEDD4","MCCC2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86YB7","full_name":"Enoyl-CoA hydratase domain-containing protein 2, mitochondrial","aliases":[],"length_aa":292,"mass_kda":31.1,"function":"","subcellular_location":"Mitochondrion","url":"https://www.uniprot.org/uniprotkb/Q86YB7/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ECHDC2","classification":"Not Classified","n_dependent_lines":21,"n_total_lines":1208,"dependency_fraction":0.0173841059602649},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ECHDC2","total_profiled":1310},"omim":[{"mim_id":"620724","title":"ENOYL COENZYME A HYDRATASE DOMAIN-CONTAINING PROTEIN 2; ECHDC2","url":"https://www.omim.org/entry/620724"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Mitochondria","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"liver","ntpm":174.8}],"url":"https://www.proteinatlas.org/search/ECHDC2"},"hgnc":{"alias_symbol":["FLJ10948"],"prev_symbol":[]},"alphafold":{"accession":"Q86YB7","domains":[{"cath_id":"3.90.226.10","chopping":"28-233","consensus_level":"high","plddt":97.8306,"start":28,"end":233},{"cath_id":"1.10.12.10","chopping":"237-284","consensus_level":"high","plddt":98.2671,"start":237,"end":284}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86YB7","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86YB7-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86YB7-F1-predicted_aligned_error_v6.png","plddt_mean":92.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ECHDC2","jax_strain_url":"https://www.jax.org/strain/search?query=ECHDC2"},"sequence":{"accession":"Q86YB7","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86YB7.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86YB7/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86YB7"}},"corpus_meta":[{"pmid":"24005033","id":"PMC_24005033","title":"Functional epigenetic approach identifies frequently methylated genes in Ewing sarcoma.","date":"2013","source":"Epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/24005033","citation_count":33,"is_preprint":false},{"pmid":"31437751","id":"PMC_31437751","title":"Multi-Omic Approaches Identify Metabolic and Autophagy Regulators Important in Ovarian Cancer Dissemination.","date":"2019","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/31437751","citation_count":26,"is_preprint":false},{"pmid":"38783226","id":"PMC_38783226","title":"ECHDC2 inhibits the proliferation of gastric cancer cells by binding with NEDD4 to degrade MCCC2 and reduce aerobic glycolysis.","date":"2024","source":"Molecular medicine (Cambridge, Mass.)","url":"https://pubmed.ncbi.nlm.nih.gov/38783226","citation_count":21,"is_preprint":false},{"pmid":"24108764","id":"PMC_24108764","title":"Enoyl coenzyme a hydratase domain-containing 2, a potential novel regulator of myocardial ischemia injury.","date":"2013","source":"Journal of the American Heart Association","url":"https://pubmed.ncbi.nlm.nih.gov/24108764","citation_count":17,"is_preprint":false},{"pmid":"36618918","id":"PMC_36618918","title":"Proteomic analysis revealed the pharmacological mechanism of Xueshuantong injection in preventing early acute myocardial infarction injury.","date":"2022","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/36618918","citation_count":8,"is_preprint":false},{"pmid":"39384885","id":"PMC_39384885","title":"Multiomics identification of ALDH9A1 as a crucial immunoregulatory molecule involved in calcific aortic valve disease.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39384885","citation_count":4,"is_preprint":false},{"pmid":"39732885","id":"PMC_39732885","title":"Genome-wide analysis of alternative splicing differences in hepatic ischemia reperfusion injury.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39732885","citation_count":2,"is_preprint":false},{"pmid":"34218094","id":"PMC_34218094","title":"Proteomic consequences of the deletion of cytochrome P450 (CYP450) reductase in mice.","date":"2021","source":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34218094","citation_count":2,"is_preprint":false},{"pmid":"32917399","id":"PMC_32917399","title":"Exploring allele specific methylation in drug dependence susceptibility.","date":"2020","source":"Journal of psychiatric research","url":"https://pubmed.ncbi.nlm.nih.gov/32917399","citation_count":1,"is_preprint":false},{"pmid":"41444657","id":"PMC_41444657","title":"Identification of coagulation-related hub genes in ischemic stroke based on bioinformatics integration analysis and investigation of their immune regulatory mechanisms.","date":"2025","source":"European journal of medical research","url":"https://pubmed.ncbi.nlm.nih.gov/41444657","citation_count":1,"is_preprint":false},{"pmid":"39409102","id":"PMC_39409102","title":"Investigating the Causal Effects of Exercise-Induced Genes on Sarcopenia.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39409102","citation_count":1,"is_preprint":false},{"pmid":"40680899","id":"PMC_40680899","title":"Very high-dose vitamin D₃ supplementation reduces the expression of genes and proteins engaged in β-oxidation in healthy pigs.","date":"2025","source":"The Journal of steroid biochemistry and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/40680899","citation_count":1,"is_preprint":false},{"pmid":"41732157","id":"PMC_41732157","title":"Clinical significance and oncogenic role of ECHDC2 in glioblastoma: a comprehensive analysis based on bioinformatics and in vitro experiments.","date":"2026","source":"Frontiers in genetics","url":"https://pubmed.ncbi.nlm.nih.gov/41732157","citation_count":0,"is_preprint":false},{"pmid":"41088987","id":"PMC_41088987","title":"Investigating the Mechanisms of Mitochondrial Dysfunction in Ischemic Stroke and Predicting Therapeutics Through Machine Learning and Integrated Bioinformatics.","date":"2025","source":"Current medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/41088987","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10054,"output_tokens":1275,"usd":0.024643,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":7972,"output_tokens":2258,"usd":0.048155,"stage2_stop_reason":"end_turn"},"total_usd":0.072798,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2013,\n      \"finding\": \"ECHDC2 is a mitochondrial protein expressed in cardiomyocytes but not cardiofibroblasts. Overexpression of ECHDC2 increased susceptibility to ischemia/reperfusion (I/R) injury, while knockdown enhanced resistance to I/R injury. ECHDC2 overexpression increased cellular levels of branched-chain amino acids leucine and valine (measured by GC-MS), but did not alter mitochondrial O2 consumption, metabolic intermediates, or ATP production.\",\n      \"method\": \"Subcellular fractionation/immunohistochemistry for localization; overexpression and knockdown with I/R injury assay; GC-MS metabolite analysis; Western blot; RT-PCR\",\n      \"journal\": \"Journal of the American Heart Association\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (localization, gain/loss-of-function, metabolomics) in a single lab; no independent replication\",\n      \"pmids\": [\"24108764\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ECHDC2 binds NEDD4 (E3 ubiquitin ligase) via co-immunoprecipitation, promotes ubiquitination and proteasomal degradation of MCCC2, and thereby suppresses the P38 MAPK pathway, leading to reduced aerobic glycolysis and inhibition of gastric cancer cell proliferation both in vitro and in vivo.\",\n      \"method\": \"Co-immunoprecipitation, Western blotting, immunofluorescence, colony formation, CCK8, EDU, glucose/lactate assays, subcutaneous xenograft in nude mice\",\n      \"journal\": \"Molecular medicine (Cambridge, Mass.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP demonstrating ECHDC2-NEDD4 interaction plus functional in vivo/in vitro validation; single lab, no independent replication\",\n      \"pmids\": [\"38783226\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"ECHDC2 knockdown significantly inhibited suspension growth (anoikis escape) of high-grade serous ovarian cancer cells, implicating ECHDC2 as an effector of fatty acid metabolism required for survival in suspension.\",\n      \"method\": \"Genome-wide CRISPR/Cas9 knockout screen followed by targeted siRNA knockdown; suspension growth assay; non-targeted metabolomics\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — GeCKO screen plus targeted knockdown validation with functional readout; single study, metabolomic support\",\n      \"pmids\": [\"31437751\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"ECHDC2 knockdown suppressed proliferation and migration of glioblastoma (GBM) cell lines, while overexpression had the opposite effect; enrichment analyses associated ECHDC2 activity with the PI3K/Akt signaling pathway in GBM.\",\n      \"method\": \"siRNA knockdown and overexpression in GBM cell lines; CCK-8, EdU, wound-healing, and Transwell migration assays; GSEA/KEGG pathway enrichment\",\n      \"journal\": \"Frontiers in genetics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, cellular phenotype established but pathway placement relies on bioinformatics enrichment without direct pathway validation\",\n      \"pmids\": [\"41732157\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Very high-dose vitamin D3 supplementation in pigs downregulated ECHDC2 protein levels in liver tissue, consistent with a role for ECHDC2 in mitochondrial β-oxidation pathways.\",\n      \"method\": \"Integrative transcriptomic and proteomic analysis of liver tissue; quantitative proteomics\",\n      \"journal\": \"The Journal of steroid biochemistry and molecular biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — protein-level change observed in a non-targeted proteomics study; no direct functional experiment on ECHDC2 itself\",\n      \"pmids\": [\"40680899\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ECHDC2 is a mitochondrial protein involved in fatty acid β-oxidation and branched-chain amino acid metabolism; it promotes ubiquitination and degradation of MCCC2 via interaction with the E3 ligase NEDD4, thereby suppressing the P38 MAPK pathway and aerobic glycolysis, and its expression level modulates susceptibility to ischemia/reperfusion injury as well as proliferation and survival in multiple cancer contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ECHDC2 is a mitochondrial protein linked to branched-chain amino acid and fatty acid metabolism that modulates cellular survival and proliferation across cardiac and cancer contexts [#0, #2]. In cardiomyocytes, ECHDC2 localizes to mitochondria, and its level sets susceptibility to ischemia/reperfusion injury—overexpression sensitizes cells while knockdown confers resistance—accompanied by elevated branched-chain amino acids leucine and valine without measurable changes in oxygen consumption or ATP production [#0]. In gastric cancer, ECHDC2 binds the E3 ubiquitin ligase NEDD4 and promotes ubiquitination and proteasomal degradation of MCCC2, thereby suppressing P38 MAPK signaling, aerobic glycolysis, and tumor cell proliferation in vitro and in xenografts [#1]. ECHDC2 is also required for anoikis escape and suspension growth of high-grade serous ovarian cancer cells, consistent with a role downstream of fatty acid metabolism [#2]. Beyond these contexts and the NEDD4–MCCC2 axis, no enzymatic substrate or structural mechanism for ECHDC2 has been characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2013,\n      \"claim\": \"Established ECHDC2 as a mitochondrial protein whose abundance bidirectionally controls cardiomyocyte vulnerability to ischemia/reperfusion injury and influences branched-chain amino acid levels.\",\n      \"evidence\": \"Subcellular fractionation/IHC localization, gain- and loss-of-function with I/R injury assay, and GC-MS metabolomics in cardiomyocytes\",\n      \"pmids\": [\"24108764\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"The enzymatic activity of ECHDC2 on BCAA or fatty acid intermediates was not directly demonstrated\",\n        \"Mechanism linking ECHDC2 to I/R susceptibility unresolved despite no change in O2 consumption or ATP\",\n        \"No in vivo cardiac model beyond cellular assays\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified ECHDC2 as a fatty-acid-metabolism effector required for anoikis escape, extending its role from cardiac biology to cancer cell survival in suspension.\",\n      \"evidence\": \"Genome-wide CRISPR/Cas9 screen with siRNA validation, suspension growth assay, and non-targeted metabolomics in ovarian cancer cells\",\n      \"pmids\": [\"31437751\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"The metabolic substrate of ECHDC2 driving suspension survival was not defined\",\n        \"No reciprocal rescue or overexpression to confirm specificity\",\n        \"Connection to a defined signaling pathway not established\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined a molecular mechanism for ECHDC2's tumor-suppressive action: recruitment of NEDD4 to drive MCCC2 degradation, dampening P38 MAPK signaling and aerobic glycolysis.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation, ubiquitination and Western assays, glucose/lactate measurements, and subcutaneous xenografts in gastric cancer\",\n      \"pmids\": [\"38783226\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether ECHDC2 directly recruits NEDD4 or acts as an adaptor versus enzyme is not resolved\",\n        \"Single lab without independent replication\",\n        \"Reconciliation with pro-survival roles in ovarian/glioblastoma contexts not addressed\"\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showed that, in contrast to its gastric tumor-suppressive role, ECHDC2 promotes proliferation and migration of glioblastoma cells, associating it with PI3K/Akt signaling.\",\n      \"evidence\": \"siRNA knockdown and overexpression with proliferation/migration assays and GSEA/KEGG enrichment in GBM cell lines\",\n      \"pmids\": [\"41732157\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"PI3K/Akt placement rests on bioinformatic enrichment without direct pathway validation\",\n        \"Context-dependent opposing roles across cancers mechanistically unexplained\",\n        \"No in vivo confirmation\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Provided correlative in vivo evidence linking ECHDC2 abundance to mitochondrial β-oxidation by showing high-dose vitamin D3 downregulates hepatic ECHDC2.\",\n      \"evidence\": \"Integrative transcriptomic and quantitative proteomic analysis of pig liver tissue\",\n      \"pmids\": [\"40680899\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Protein-level change observed without any direct functional test of ECHDC2\",\n        \"Causal relationship between vitamin D3 and ECHDC2 not established\",\n        \"β-oxidation role inferred, not measured\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The direct biochemical activity of ECHDC2 and how it produces opposing proliferative outcomes across tissues remain unknown.\",\n      \"evidence\": \"No reconstituted enzymatic assay or structural characterization in the available corpus\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No defined catalytic substrate or reaction for ECHDC2\",\n        \"Mechanism reconciling tumor-suppressor (gastric) versus pro-tumor (GBM) roles unresolved\",\n        \"No structural model or domain-level mechanism of NEDD4/MCCC2 engagement\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [0, 2]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"NEDD4\", \"MCCC2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":4,"faith_total":4,"faith_pct":100.0}}