{"gene":"EPHA10","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":2005,"finding":"EphA10 is a receptor tyrosine kinase expressed predominantly in testis; three isoforms were identified (one soluble, two transmembrane, one of which lacks the SAM domain). Ligand-binding studies demonstrated that EphA10 binds preferentially to ephrin-A ligands, classifying it in the EphA subclass.","method":"Isoform identification by molecular cloning; ephrin ligand binding studies","journal":"Biochimica et biophysica acta","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — direct binding assay and isoform characterization in a single focused study, but no mutagenesis or structural validation","pmids":["15777695"],"is_preprint":false},{"year":2016,"finding":"EPHA10 physically interacts with the kinase-sufficient EPHA7 receptor, as demonstrated by co-immunoprecipitation. The two receptors co-localize on the cell surface; soluble isoforms form a complex in the cytoplasm and nucleus of breast carcinoma cells, suggesting a gene-regulatory function for the nuclear complex.","method":"Co-immunoprecipitation; immunocytochemistry/confocal microscopy","journal":"Cancer genomics & proteomics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — reciprocal Co-IP plus orthogonal immunocytochemical localization in a single lab study","pmids":["27566654"],"is_preprint":false},{"year":2017,"finding":"EPHA10 isoform expression patterns regulate breast cancer progression: the soluble secretory isoform EphA10s stabilizes membrane-associated β-catenin via interaction with ephrin-A5, while the cytoplasmic full-length isoform maintains phosphorylation of E-cadherin. Restoring the normal isoform balance (up-regulating EphA10s, down-regulating cytoplasmic EphA10) strengthened the E-cadherin/β-catenin membrane complex and inhibited cell invasion and lymph node metastasis.","method":"Isoform-specific overexpression and knockdown; co-immunoprecipitation for EphA10s–ephrin-A5 interaction; Western blot for E-cadherin phosphorylation; invasion assays; in vivo metastasis model","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — multiple functional readouts and molecular measurements in a single lab, but no structural or in vitro reconstitution data","pmids":["28427223"],"is_preprint":false},{"year":2020,"finding":"EphA10 (a catalytically defective RTK) promotes tumorigenesis in pancreatic cancer cells by increasing phosphorylation of ERK, JNK, AKT, FAK, and NF-κB, and by enhancing expression and secretion of MMP-9. EphA10 silencing reduced proliferation, migration, and adhesion, whereas overexpression reversed these effects and increased vascular density in xenograft tumors.","method":"siRNA knockdown and cDNA overexpression; Western blot for downstream signaling; gelatin degradation/invasion assays; MIA PaCa-2 xenograft model","journal":"Cancer science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO/OE with defined cellular and in vivo phenotypes and multiple downstream signaling readouts, single lab","pmids":["32644283"],"is_preprint":false},{"year":2021,"finding":"The intracellular region of EphA10 (juxtamembrane region, pseudokinase domain, and SAM domain) is highly flexible in solution and shows interdomain interactions, as determined by small-angle X-ray scattering and cross-linking mass spectrometry. EphA10's pseudokinase domain can bind ATP and ATP-competitive small molecules, indicating the domain is pharmacologically tractable despite catalytic inactivity.","method":"Small-angle X-ray scattering (SAXS); cross-linking mass spectrometry; ATP-binding assays","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1 / Moderate — biophysical structural characterization (SAXS) combined with cross-linking MS and direct ATP-binding assay in a single rigorous biochemical study","pmids":["34431498"],"is_preprint":false},{"year":2022,"finding":"EphA10 activates the MAPK/ERK pathway in lung adenocarcinoma cells; pharmacological inhibition of MEK with U0126 reversed the pro-tumorigenic effects of EphA10 overexpression, establishing EphA10 acts upstream of MEK/ERK. EphA10 knockdown also reduced PD-L1 expression, enhancing NK cell-mediated anti-tumor activity.","method":"Lentiviral knockdown/overexpression; Western blot; MEK inhibitor (U0126) epistasis; co-culture NK cell cytotoxicity assay; xenograft model","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis via MEK inhibitor combined with KO/OE and defined signaling readouts, single lab","pmids":["35839564"],"is_preprint":false},{"year":2023,"finding":"EPHA10 is expressed in the mouse cochlea at both mRNA and protein levels. Overexpression of the Drosophila homolog of EPHA10 (Eph) disrupted the structure and function of chordotonal organs in fly models, and a 5′-UTR non-coding variant that upregulates EPHA10 expression co-segregated with autosomal dominant non-syndromic hearing loss, linking EPHA10 dosage to auditory function.","method":"Mouse cochlear expression by RT-PCR and immunostaining; Drosophila Eph overexpression functional assay; variant-driven promoter activity assay; family-based co-segregation analysis","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo functional model (fly) combined with promoter activity assay and expression localization, single study","pmids":["36048850"],"is_preprint":false},{"year":2025,"finding":"EphA10 mRNA undergoes N6-methyladenosine (m6A) modification written by RBM15B and read by YTHDF1, which stabilizes EphA10 mRNA and enhances its expression. m6A-modified EphA10 activates the ERK/AKT signaling pathway to promote prostate cancer cell proliferation, invasion, and migration.","method":"Dot blot and MeRIP-qPCR for m6A modification; siRNA knockdown of RBM15B and YTHDF1; Western blot and qRT-PCR; functional proliferation/invasion/migration assays","journal":"Biochemical genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct m6A detection by MeRIP-qPCR plus writer/reader knockdown, single lab, single study","pmids":["41296142"],"is_preprint":false}],"current_model":"EphA10 is a catalytically defective (pseudokinase) EphA-subclass receptor that preferentially binds ephrin-A ligands, possesses a flexible intracellular region capable of ATP binding, and signals non-catalytically through interactions with active Eph receptors (e.g., EPHA7) and by scaffolding downstream activation of ERK, JNK, AKT, FAK, and NF-κB pathways; its expression is post-transcriptionally regulated by RBM15B-mediated m6A methylation read by YTHDF1, and isoform-specific localization (secreted vs. cytoplasmic) differentially controls E-cadherin/β-catenin complex stability and cell invasiveness."},"narrative":{"mechanistic_narrative":"EPHA10 is a catalytically defective (pseudokinase) EphA-subclass receptor that preferentially binds ephrin-A ligands and contributes to epithelial adhesion and tumor cell behavior through non-catalytic signaling [PMID:15777695, PMID:32644283]. Although its intracellular region lacks catalytic activity, the juxtamembrane–pseudokinase–SAM module is conformationally flexible and retains the ability to bind ATP and ATP-competitive small molecules, making the pseudokinase domain pharmacologically tractable [PMID:34431498]. EphA10 physically associates with the kinase-competent receptor EPHA7, co-localizing at the cell surface and, for soluble isoforms, forming complexes in the cytoplasm and nucleus [PMID:27566654]. Functionally, EphA10 isoforms differentially control the E-cadherin/β-catenin membrane complex: the secreted EphA10s isoform stabilizes membrane β-catenin via ephrin-A5 interaction, while the cytoplasmic full-length isoform maintains E-cadherin phosphorylation, and restoring the normal isoform balance suppresses invasion and metastasis [PMID:28427223]. In multiple cancers EphA10 acts as a scaffold driving pro-tumorigenic signaling — increasing phosphorylation of ERK, JNK, AKT, FAK, and NF-κB and enhancing MMP-9 secretion in pancreatic cells [PMID:32644283], acting upstream of MEK/ERK and promoting PD-L1 expression in lung adenocarcinoma [PMID:35839564]. EphA10 expression is post-transcriptionally controlled by RBM15B-written m6A modification read by YTHDF1, which stabilizes its mRNA and activates ERK/AKT signaling in prostate cancer [PMID:41296142]. A 5′-UTR variant that upregulates EPHA10 co-segregates with autosomal dominant non-syndromic hearing loss, linking EPHA10 dosage to auditory function [PMID:36048850].","teleology":[{"year":2005,"claim":"Establishing EPHA10 as a receptor with ligand-binding identity answered whether this orphan-like gene was a functional Eph receptor and to which subclass it belonged.","evidence":"Molecular cloning of three isoforms and ephrin ligand-binding assays in testis-derived material","pmids":["15777695"],"confidence":"Medium","gaps":["No mutagenesis or structural validation of the ligand-binding interface","Catalytic status not yet established at this stage","Functional consequence of ephrin binding not defined"]},{"year":2016,"claim":"Identifying a physical interaction with the kinase-competent EPHA7 addressed how a catalytically defective receptor could signal, pointing to heteromeric receptor coupling.","evidence":"Reciprocal co-immunoprecipitation and confocal immunocytochemistry in breast carcinoma cells","pmids":["27566654"],"confidence":"Medium","gaps":["Single-lab study without structural mapping of the interaction","Proposed nuclear gene-regulatory function not mechanistically defined","No demonstration that EPHA7 transphosphorylates or activates EphA10"]},{"year":2017,"claim":"Dissecting isoform-specific roles answered how EphA10 controls epithelial integrity, showing secreted versus cytoplasmic isoforms oppositely regulate the E-cadherin/β-catenin complex and invasiveness.","evidence":"Isoform-specific overexpression/knockdown, Co-IP of EphA10s–ephrin-A5, E-cadherin phosphorylation Westerns, invasion assays, and an in vivo metastasis model","pmids":["28427223"],"confidence":"Medium","gaps":["No structural or in vitro reconstitution of the EphA10s–ephrin-A5 complex","Mechanism by which the cytoplasmic isoform maintains E-cadherin phosphorylation unresolved","Confined to breast cancer context"]},{"year":2020,"claim":"Defining downstream signaling answered how EphA10 promotes tumorigenesis non-catalytically, linking it to multiple kinase cascades and matrix remodeling.","evidence":"siRNA knockdown and overexpression with phospho-Westerns (ERK/JNK/AKT/FAK/NF-κB), gelatin degradation assays, and MIA PaCa-2 xenografts","pmids":["32644283"],"confidence":"Medium","gaps":["Direct molecular link between EphA10 and each phosphorylated effector not established","Scaffolding mechanism vs indirect effect not distinguished","Single lineage (pancreatic cells)"]},{"year":2021,"claim":"Biophysical characterization answered whether the pseudokinase domain is a viable drug target despite catalytic inactivity, revealing a flexible ATP-binding module.","evidence":"SAXS, cross-linking mass spectrometry, and direct ATP / ATP-competitive small-molecule binding assays of the intracellular region","pmids":["34431498"],"confidence":"High","gaps":["No high-resolution crystal/cryo-EM structure","Functional role of ATP binding in signaling not determined","Interdomain interactions not linked to cellular activity"]},{"year":2022,"claim":"Epistasis testing placed EphA10 upstream of MEK/ERK and connected it to immune evasion, clarifying its position in the signaling hierarchy.","evidence":"Lentiviral knockdown/overexpression with U0126 MEK-inhibitor epistasis, NK-cell co-culture cytotoxicity, and xenografts in lung adenocarcinoma","pmids":["35839564"],"confidence":"Medium","gaps":["Mechanism linking EphA10 to PD-L1 expression undefined","Direct effector between EphA10 and MEK not identified","Single tumor type"]},{"year":2023,"claim":"Demonstrating cochlear expression and a dosage-sensitive variant answered whether EPHA10 has a physiological role beyond cancer, implicating it in hearing.","evidence":"Mouse cochlear RT-PCR/immunostaining, Drosophila Eph overexpression chordotonal-organ assay, promoter-activity assay of a 5′-UTR variant, and family co-segregation","pmids":["36048850"],"confidence":"Medium","gaps":["Mechanism connecting EPHA10 overexpression to auditory dysfunction unknown","Reliance on a Drosophila homolog rather than the mammalian gene for functional readout","Single family/study"]},{"year":2025,"claim":"Identifying m6A regulation answered how EphA10 expression is controlled post-transcriptionally, defining an RBM15B/YTHDF1 axis that stabilizes its mRNA.","evidence":"Dot blot and MeRIP-qPCR for m6A, RBM15B/YTHDF1 knockdown, and proliferation/invasion/migration assays in prostate cancer cells","pmids":["41296142"],"confidence":"Medium","gaps":["Specific m6A sites on EphA10 mRNA not mapped","Direct binding of YTHDF1 to EphA10 transcript not shown","Single lab, single study"]},{"year":null,"claim":"Whether the EphA10 pseudokinase actively transmits signal through EPHA7 heterodimers in a defined biochemical pathway, and how this couples to the diverse downstream cascades, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No reconstituted signaling complex linking EPHA7 binding to ERK/AKT/JNK/FAK/NF-κB activation","No structure of the receptor in complex with ligand or EPHA7","Physiological (non-cancer) signaling mechanism in testis and cochlea undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[4]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,1]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,5]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,2]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[1,2]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[1]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[0,2]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,5]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[2,3,5,7]},{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[7]}],"complexes":[],"partners":["EPHA7","EFNA5","RBM15B","YTHDF1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q5JZY3","full_name":"Ephrin type-A receptor 10","aliases":[],"length_aa":1008,"mass_kda":109.7,"function":"Receptor for members of the ephrin-A family. Binds to EFNA3, EFNA4 and EFNA5","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/Q5JZY3/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EPHA10","classification":"Not Classified","n_dependent_lines":6,"n_total_lines":1208,"dependency_fraction":0.004966887417218543},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/EPHA10","total_profiled":1310},"omim":[{"mim_id":"620283","title":"DEAFNESS, AUTOSOMAL DOMINANT 88; DFNA88","url":"https://www.omim.org/entry/620283"},{"mim_id":"611123","title":"EPHRIN RECEPTOR EphA10; EPHA10","url":"https://www.omim.org/entry/611123"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"brain","ntpm":8.9},{"tissue":"intestine","ntpm":12.5},{"tissue":"testis","ntpm":12.4}],"url":"https://www.proteinatlas.org/search/EPHA10"},"hgnc":{"alias_symbol":["FLJ16103","FLJ33655"],"prev_symbol":[]},"alphafold":{"accession":"Q5JZY3","domains":[{"cath_id":"2.60.120.260","chopping":"35-212","consensus_level":"high","plddt":84.4518,"start":35,"end":212},{"cath_id":"2.60.40.1770","chopping":"215-272","consensus_level":"medium","plddt":85.621,"start":215,"end":272},{"cath_id":"2.60.40.10","chopping":"342-450","consensus_level":"medium","plddt":82.6442,"start":342,"end":450},{"cath_id":"2.60.40.10","chopping":"461-475_489-498_505-536_544-552","consensus_level":"medium","plddt":83.2715,"start":461,"end":552},{"cath_id":"3.30.200.20","chopping":"640-725","consensus_level":"high","plddt":88.682,"start":640,"end":725},{"cath_id":"1.10.510.10","chopping":"730-906","consensus_level":"high","plddt":88.659,"start":730,"end":906},{"cath_id":"1.10.150.50","chopping":"939-1004","consensus_level":"high","plddt":80.8976,"start":939,"end":1004}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q5JZY3","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q5JZY3-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q5JZY3-F1-predicted_aligned_error_v6.png","plddt_mean":79.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EPHA10","jax_strain_url":"https://www.jax.org/strain/search?query=EPHA10"},"sequence":{"accession":"Q5JZY3","fasta_url":"https://rest.uniprot.org/uniprotkb/Q5JZY3.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q5JZY3/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q5JZY3"}},"corpus_meta":[{"pmid":"15777695","id":"PMC_15777695","title":"Characterization of a novel Eph receptor tyrosine kinase, EphA10, expressed in testis.","date":"2005","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/15777695","citation_count":54,"is_preprint":false},{"pmid":"29798663","id":"PMC_29798663","title":"Overcoming Multidrug Resistance by Codelivery of MDR1-Targeting siRNA and Doxorubicin Using EphA10-Mediated pH-Sensitive Lipoplexes: In Vitro and In Vivo Evaluation.","date":"2018","source":"ACS applied materials & interfaces","url":"https://pubmed.ncbi.nlm.nih.gov/29798663","citation_count":50,"is_preprint":false},{"pmid":"27574425","id":"PMC_27574425","title":"Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA.","date":"2016","source":"International journal of nanomedicine","url":"https://pubmed.ncbi.nlm.nih.gov/27574425","citation_count":30,"is_preprint":false},{"pmid":"32644283","id":"PMC_32644283","title":"The catalytically defective receptor protein tyrosine kinase EphA10 promotes tumorigenesis in pancreatic cancer cells.","date":"2020","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/32644283","citation_count":19,"is_preprint":false},{"pmid":"28427223","id":"PMC_28427223","title":"Isoform expression patterns of EPHA10 protein mediate breast cancer progression by regulating the E-Cadherin and β-catenin complex.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/28427223","citation_count":16,"is_preprint":false},{"pmid":"27566654","id":"PMC_27566654","title":"EPHA7 and EPHA10 Physically Interact and Differentially Co-localize in Normal Breast and Breast Carcinoma Cell Lines, and the Co-localization Pattern Is Altered in EPHB6-expressing MDA-MB-231 Cells.","date":"2016","source":"Cancer genomics & proteomics","url":"https://pubmed.ncbi.nlm.nih.gov/27566654","citation_count":15,"is_preprint":false},{"pmid":"34431498","id":"PMC_34431498","title":"The intracellular domains of the EphB6 and EphA10 receptor tyrosine pseudokinases function as dynamic signalling hubs.","date":"2021","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/34431498","citation_count":13,"is_preprint":false},{"pmid":"35839564","id":"PMC_35839564","title":"EphA10 drives tumor progression and immune evasion by regulating the MAPK/ERK cascade in lung adenocarcinoma.","date":"2022","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/35839564","citation_count":13,"is_preprint":false},{"pmid":"33990369","id":"PMC_33990369","title":"Coexpression of EphA10 and Gli3 promotes breast cancer cell proliferation, invasion and migration.","date":"2021","source":"Journal of investigative medicine : the official publication of the American Federation for Clinical Research","url":"https://pubmed.ncbi.nlm.nih.gov/33990369","citation_count":7,"is_preprint":false},{"pmid":"36048850","id":"PMC_36048850","title":"A non-coding variant in 5' untranslated region drove up-regulation of pseudo-kinase EPHA10 and caused non-syndromic hearing loss in humans.","date":"2023","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/36048850","citation_count":4,"is_preprint":false},{"pmid":"41296142","id":"PMC_41296142","title":"The N6-methyladenosine Modified EphA10 Promotes Prostate Cancer Progression by Activating the ERK/AKT Pathway.","date":"2025","source":"Biochemical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/41296142","citation_count":0,"is_preprint":false},{"pmid":"39710122","id":"PMC_39710122","title":"The mechanism of all-trans retinoic acid-induced cleft palate may be related to the novel ENSMUST00000159153-miR-137-5p-Wnt7a and ENSMUST000000236086-miR-34b-3p-EphA10/TRPM2 ceRNA crosstalk.","date":"2024","source":"Environmental toxicology and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/39710122","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.04.23.650160","title":"A New Serological Autoantibody Signature Associated with Multiple Sclerosis","date":"2025-04-26","source":"bioRxiv","url":"https://doi.org/10.1101/2025.04.23.650160","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":8370,"output_tokens":2206,"usd":0.0291,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9221,"output_tokens":3386,"usd":0.065378,"stage2_stop_reason":"end_turn"},"total_usd":0.094478,"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\": 2005,\n      \"finding\": \"EphA10 is a receptor tyrosine kinase expressed predominantly in testis; three isoforms were identified (one soluble, two transmembrane, one of which lacks the SAM domain). Ligand-binding studies demonstrated that EphA10 binds preferentially to ephrin-A ligands, classifying it in the EphA subclass.\",\n      \"method\": \"Isoform identification by molecular cloning; ephrin ligand binding studies\",\n      \"journal\": \"Biochimica et biophysica acta\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — direct binding assay and isoform characterization in a single focused study, but no mutagenesis or structural validation\",\n      \"pmids\": [\"15777695\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"EPHA10 physically interacts with the kinase-sufficient EPHA7 receptor, as demonstrated by co-immunoprecipitation. The two receptors co-localize on the cell surface; soluble isoforms form a complex in the cytoplasm and nucleus of breast carcinoma cells, suggesting a gene-regulatory function for the nuclear complex.\",\n      \"method\": \"Co-immunoprecipitation; immunocytochemistry/confocal microscopy\",\n      \"journal\": \"Cancer genomics & proteomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — reciprocal Co-IP plus orthogonal immunocytochemical localization in a single lab study\",\n      \"pmids\": [\"27566654\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"EPHA10 isoform expression patterns regulate breast cancer progression: the soluble secretory isoform EphA10s stabilizes membrane-associated β-catenin via interaction with ephrin-A5, while the cytoplasmic full-length isoform maintains phosphorylation of E-cadherin. Restoring the normal isoform balance (up-regulating EphA10s, down-regulating cytoplasmic EphA10) strengthened the E-cadherin/β-catenin membrane complex and inhibited cell invasion and lymph node metastasis.\",\n      \"method\": \"Isoform-specific overexpression and knockdown; co-immunoprecipitation for EphA10s–ephrin-A5 interaction; Western blot for E-cadherin phosphorylation; invasion assays; in vivo metastasis model\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — multiple functional readouts and molecular measurements in a single lab, but no structural or in vitro reconstitution data\",\n      \"pmids\": [\"28427223\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EphA10 (a catalytically defective RTK) promotes tumorigenesis in pancreatic cancer cells by increasing phosphorylation of ERK, JNK, AKT, FAK, and NF-κB, and by enhancing expression and secretion of MMP-9. EphA10 silencing reduced proliferation, migration, and adhesion, whereas overexpression reversed these effects and increased vascular density in xenograft tumors.\",\n      \"method\": \"siRNA knockdown and cDNA overexpression; Western blot for downstream signaling; gelatin degradation/invasion assays; MIA PaCa-2 xenograft model\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO/OE with defined cellular and in vivo phenotypes and multiple downstream signaling readouts, single lab\",\n      \"pmids\": [\"32644283\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"The intracellular region of EphA10 (juxtamembrane region, pseudokinase domain, and SAM domain) is highly flexible in solution and shows interdomain interactions, as determined by small-angle X-ray scattering and cross-linking mass spectrometry. EphA10's pseudokinase domain can bind ATP and ATP-competitive small molecules, indicating the domain is pharmacologically tractable despite catalytic inactivity.\",\n      \"method\": \"Small-angle X-ray scattering (SAXS); cross-linking mass spectrometry; ATP-binding assays\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — biophysical structural characterization (SAXS) combined with cross-linking MS and direct ATP-binding assay in a single rigorous biochemical study\",\n      \"pmids\": [\"34431498\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"EphA10 activates the MAPK/ERK pathway in lung adenocarcinoma cells; pharmacological inhibition of MEK with U0126 reversed the pro-tumorigenic effects of EphA10 overexpression, establishing EphA10 acts upstream of MEK/ERK. EphA10 knockdown also reduced PD-L1 expression, enhancing NK cell-mediated anti-tumor activity.\",\n      \"method\": \"Lentiviral knockdown/overexpression; Western blot; MEK inhibitor (U0126) epistasis; co-culture NK cell cytotoxicity assay; xenograft model\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis via MEK inhibitor combined with KO/OE and defined signaling readouts, single lab\",\n      \"pmids\": [\"35839564\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EPHA10 is expressed in the mouse cochlea at both mRNA and protein levels. Overexpression of the Drosophila homolog of EPHA10 (Eph) disrupted the structure and function of chordotonal organs in fly models, and a 5′-UTR non-coding variant that upregulates EPHA10 expression co-segregated with autosomal dominant non-syndromic hearing loss, linking EPHA10 dosage to auditory function.\",\n      \"method\": \"Mouse cochlear expression by RT-PCR and immunostaining; Drosophila Eph overexpression functional assay; variant-driven promoter activity assay; family-based co-segregation analysis\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo functional model (fly) combined with promoter activity assay and expression localization, single study\",\n      \"pmids\": [\"36048850\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EphA10 mRNA undergoes N6-methyladenosine (m6A) modification written by RBM15B and read by YTHDF1, which stabilizes EphA10 mRNA and enhances its expression. m6A-modified EphA10 activates the ERK/AKT signaling pathway to promote prostate cancer cell proliferation, invasion, and migration.\",\n      \"method\": \"Dot blot and MeRIP-qPCR for m6A modification; siRNA knockdown of RBM15B and YTHDF1; Western blot and qRT-PCR; functional proliferation/invasion/migration assays\",\n      \"journal\": \"Biochemical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct m6A detection by MeRIP-qPCR plus writer/reader knockdown, single lab, single study\",\n      \"pmids\": [\"41296142\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EphA10 is a catalytically defective (pseudokinase) EphA-subclass receptor that preferentially binds ephrin-A ligands, possesses a flexible intracellular region capable of ATP binding, and signals non-catalytically through interactions with active Eph receptors (e.g., EPHA7) and by scaffolding downstream activation of ERK, JNK, AKT, FAK, and NF-κB pathways; its expression is post-transcriptionally regulated by RBM15B-mediated m6A methylation read by YTHDF1, and isoform-specific localization (secreted vs. cytoplasmic) differentially controls E-cadherin/β-catenin complex stability and cell invasiveness.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EPHA10 is a catalytically defective (pseudokinase) EphA-subclass receptor that preferentially binds ephrin-A ligands and contributes to epithelial adhesion and tumor cell behavior through non-catalytic signaling [#0, #3]. Although its intracellular region lacks catalytic activity, the juxtamembrane–pseudokinase–SAM module is conformationally flexible and retains the ability to bind ATP and ATP-competitive small molecules, making the pseudokinase domain pharmacologically tractable [#4]. EphA10 physically associates with the kinase-competent receptor EPHA7, co-localizing at the cell surface and, for soluble isoforms, forming complexes in the cytoplasm and nucleus [#1]. Functionally, EphA10 isoforms differentially control the E-cadherin/β-catenin membrane complex: the secreted EphA10s isoform stabilizes membrane β-catenin via ephrin-A5 interaction, while the cytoplasmic full-length isoform maintains E-cadherin phosphorylation, and restoring the normal isoform balance suppresses invasion and metastasis [#2]. In multiple cancers EphA10 acts as a scaffold driving pro-tumorigenic signaling — increasing phosphorylation of ERK, JNK, AKT, FAK, and NF-κB and enhancing MMP-9 secretion in pancreatic cells [#3], acting upstream of MEK/ERK and promoting PD-L1 expression in lung adenocarcinoma [#5]. EphA10 expression is post-transcriptionally controlled by RBM15B-written m6A modification read by YTHDF1, which stabilizes its mRNA and activates ERK/AKT signaling in prostate cancer [#7]. A 5′-UTR variant that upregulates EPHA10 co-segregates with autosomal dominant non-syndromic hearing loss, linking EPHA10 dosage to auditory function [#6].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Establishing EPHA10 as a receptor with ligand-binding identity answered whether this orphan-like gene was a functional Eph receptor and to which subclass it belonged.\",\n      \"evidence\": \"Molecular cloning of three isoforms and ephrin ligand-binding assays in testis-derived material\",\n      \"pmids\": [\"15777695\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No mutagenesis or structural validation of the ligand-binding interface\", \"Catalytic status not yet established at this stage\", \"Functional consequence of ephrin binding not defined\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identifying a physical interaction with the kinase-competent EPHA7 addressed how a catalytically defective receptor could signal, pointing to heteromeric receptor coupling.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation and confocal immunocytochemistry in breast carcinoma cells\",\n      \"pmids\": [\"27566654\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab study without structural mapping of the interaction\", \"Proposed nuclear gene-regulatory function not mechanistically defined\", \"No demonstration that EPHA7 transphosphorylates or activates EphA10\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Dissecting isoform-specific roles answered how EphA10 controls epithelial integrity, showing secreted versus cytoplasmic isoforms oppositely regulate the E-cadherin/β-catenin complex and invasiveness.\",\n      \"evidence\": \"Isoform-specific overexpression/knockdown, Co-IP of EphA10s–ephrin-A5, E-cadherin phosphorylation Westerns, invasion assays, and an in vivo metastasis model\",\n      \"pmids\": [\"28427223\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural or in vitro reconstitution of the EphA10s–ephrin-A5 complex\", \"Mechanism by which the cytoplasmic isoform maintains E-cadherin phosphorylation unresolved\", \"Confined to breast cancer context\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defining downstream signaling answered how EphA10 promotes tumorigenesis non-catalytically, linking it to multiple kinase cascades and matrix remodeling.\",\n      \"evidence\": \"siRNA knockdown and overexpression with phospho-Westerns (ERK/JNK/AKT/FAK/NF-κB), gelatin degradation assays, and MIA PaCa-2 xenografts\",\n      \"pmids\": [\"32644283\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular link between EphA10 and each phosphorylated effector not established\", \"Scaffolding mechanism vs indirect effect not distinguished\", \"Single lineage (pancreatic cells)\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Biophysical characterization answered whether the pseudokinase domain is a viable drug target despite catalytic inactivity, revealing a flexible ATP-binding module.\",\n      \"evidence\": \"SAXS, cross-linking mass spectrometry, and direct ATP / ATP-competitive small-molecule binding assays of the intracellular region\",\n      \"pmids\": [\"34431498\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No high-resolution crystal/cryo-EM structure\", \"Functional role of ATP binding in signaling not determined\", \"Interdomain interactions not linked to cellular activity\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Epistasis testing placed EphA10 upstream of MEK/ERK and connected it to immune evasion, clarifying its position in the signaling hierarchy.\",\n      \"evidence\": \"Lentiviral knockdown/overexpression with U0126 MEK-inhibitor epistasis, NK-cell co-culture cytotoxicity, and xenografts in lung adenocarcinoma\",\n      \"pmids\": [\"35839564\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking EphA10 to PD-L1 expression undefined\", \"Direct effector between EphA10 and MEK not identified\", \"Single tumor type\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrating cochlear expression and a dosage-sensitive variant answered whether EPHA10 has a physiological role beyond cancer, implicating it in hearing.\",\n      \"evidence\": \"Mouse cochlear RT-PCR/immunostaining, Drosophila Eph overexpression chordotonal-organ assay, promoter-activity assay of a 5′-UTR variant, and family co-segregation\",\n      \"pmids\": [\"36048850\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism connecting EPHA10 overexpression to auditory dysfunction unknown\", \"Reliance on a Drosophila homolog rather than the mammalian gene for functional readout\", \"Single family/study\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identifying m6A regulation answered how EphA10 expression is controlled post-transcriptionally, defining an RBM15B/YTHDF1 axis that stabilizes its mRNA.\",\n      \"evidence\": \"Dot blot and MeRIP-qPCR for m6A, RBM15B/YTHDF1 knockdown, and proliferation/invasion/migration assays in prostate cancer cells\",\n      \"pmids\": [\"41296142\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Specific m6A sites on EphA10 mRNA not mapped\", \"Direct binding of YTHDF1 to EphA10 transcript not shown\", \"Single lab, single study\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Whether the EphA10 pseudokinase actively transmits signal through EPHA7 heterodimers in a defined biochemical pathway, and how this couples to the diverse downstream cascades, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No reconstituted signaling complex linking EPHA7 binding to ERK/AKT/JNK/FAK/NF-κB activation\", \"No structure of the receptor in complex with ligand or EPHA7\", \"Physiological (non-cancer) signaling mechanism in testis and cochlea undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 2]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1, 2]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0, 2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 5]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [2, 3, 5, 7]},\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"EPHA7\", \"EFNA5\", \"RBM15B\", \"YTHDF1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}