{"gene":"EPHA3","run_date":"2026-04-28T17:46:03","timeline":{"discoveries":[{"year":2006,"finding":"EphA3 undergoes a cis interaction with ephrinA5 that is independent of the established ligand-binding domain of EphA3; this cis interaction abolishes tyrosine phosphorylation of EphA3 and results in loss of sensitivity of retinal axons to ephrinAs in trans, effectively silencing EphA3 signaling.","method":"Truncated EphA3 constructs, single-amino acid point mutants of ephrinA5, fluorescence resonance energy transfer (FRET), and functional retinal axon assays","journal":"Nature neuroscience","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal methods (FRET, point mutagenesis, functional axon assays) in a single rigorous study","pmids":["16491080"],"is_preprint":false},{"year":2008,"finding":"The crystal structure of the EphA3 kinase domain revealed that the unphosphorylated juxtamembrane region blocks catalysis by interacting with the active site; autophosphorylation of two juxtamembrane tyrosines relieves this inhibition, and a conserved coupled pathway of residues connects the juxtamembrane segment, activation loop, and catalytic residues.","method":"High-resolution X-ray crystallography of wild-type and site-specific mutants of EphA3 kinase domain with and without juxtamembrane segment; kinase activity assays","journal":"Structure","confidence":"High","confidence_rationale":"Tier 1 — crystal structures of multiple states combined with mutagenesis and kinase activity assays","pmids":["18547520"],"is_preprint":false},{"year":2003,"finding":"Three EphA3 surface areas are essential for EphA3/ephrin-A5 recognition and assembly of signaling-competent oligomeric receptor-ligand complexes; two map to previously identified Eph/ephrin interfaces and a third is a novel contact area outside the structurally characterized interaction domains.","method":"Random mutagenesis of EphA3 expressed in yeast, selection for impaired ephrin-A5 binding, functional characterization of point mutants","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 — functional mutagenesis screen with rigorous characterization of binding and signaling","pmids":["14660665"],"is_preprint":false},{"year":2006,"finding":"EphA3 null mice exhibit significant defects in atrial septa and atrioventricular endocardial cushion development; EphA3 and its ligand ephrin-A1 are expressed in adjacent cells in the developing endocardial cushions, and EphA3-/- endocardial cushion explants produce fewer migrating mesenchymal cells, indicating EphA3 signaling drives epithelial-to-mesenchymal transformation during heart development.","method":"EphA3 knockout mouse phenotypic analysis, endocardial cushion explant migration assays, expression analysis of EphA3 and ephrin-A1","journal":"Developmental biology","confidence":"High","confidence_rationale":"Tier 2 — clean KO with defined cellular phenotype (EMT defect) and mechanistic localization of ligand-receptor pair","pmids":["17046737"],"is_preprint":false},{"year":2013,"finding":"EphA3 is highly expressed on tumor-initiating cells in glioblastoma and maintains cells in a less differentiated state by modulating MAPK signaling; EphA3 knockdown or depletion of EphA3-positive cells reduced tumorigenic potential.","method":"EphA3 knockdown, depletion of EphA3-positive tumor cells, radiolabelled EphA3-specific monoclonal antibody treatment, MAPK signaling analysis in GBM","journal":"Cancer cell","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function with specific tumorigenic and signaling readouts, multiple approaches","pmids":["23410976"],"is_preprint":false},{"year":2012,"finding":"Multiple EphA3 somatic mutations found in lung, colorectal, and hepatocellular cancers impair kinase activity, ephrin ligand binding, and/or decrease receptor cell surface localization, suggesting EphA3 has ephrin- and kinase-dependent tumor-suppressing activities disrupted by cancer mutations.","method":"Kinase activity assays, ephrin binding assays, cell surface localization analysis of mutant receptors","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal biochemical assays on cancer-associated mutants","pmids":["22242939"],"is_preprint":false},{"year":2012,"finding":"At least two cancer-associated EPHA3 somatic mutations function as dominant inhibitors of wild-type EPHA3 protein; wild-type EPHA3 re-expression in lung cancer lines increased apoptosis through suppression of AKT activation and inhibited tumor xenograft growth.","method":"Immunoprecipitation, western blotting, kinase assays, dominant-negative analysis, xenograft tumor growth assays, apoptosis assays","journal":"Journal of the National Cancer Institute","confidence":"High","confidence_rationale":"Tier 1-2 — in vitro kinase assays and in vivo xenograft experiments with mechanistic AKT pathway readout","pmids":["22829656"],"is_preprint":false},{"year":2019,"finding":"Presenilin-1/γ-secretase cleaves EphA3 independently of ephrin ligand binding, generating an intracellular domain (ICD) fragment; EphA3 ICD binds non-muscle myosin IIA (NMIIA) and increases its phosphorylation at Ser1943, promoting NMIIA filament disassembly and cytoskeleton rearrangement, thereby driving axon elongation through inhibition of RhoA signaling.","method":"Genetic PS1/EphA3 knockout neurons, proteomic analysis of EphA3 ICD interactors, phosphorylation assays, rescue experiments with ICD fragment, pharmacological NMII inhibition","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal methods including proteomics, KO genetics, rescue experiments, and phosphorylation assays","pmids":["31577226"],"is_preprint":false},{"year":2018,"finding":"Neurocan (a perineuronal net component) binds NCAM at the EphA3-binding site within the immunoglobulin-2 domain, inhibiting NCAM/EphA3 association, membrane clustering of NCAM/EphA3 in cortical interneuron axons, EphA3 kinase activation, and ephrin-A5-induced growth cone collapse.","method":"Binding assays, co-immunoprecipitation, EphA3 kinase activation assays, growth cone collapse assays in cortical interneuron cultures, organotypic cortical slice cultures with enzymatic PNN removal","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods including binding assays, kinase assays, and functional growth cone assays","pmids":["29670169"],"is_preprint":false},{"year":2014,"finding":"Hypoxia-inducible factor (HIF)-1α controls EphA3 expression in human endometrial multipotent mesenchymal stromal cells (eMSCs); EphA3 agonists induce receptor phosphorylation, cell contraction, cell-cell segregation and directed cell migration, and EphA3 silencing significantly inhibits transplanted eMSC support of neovascularisation in vivo.","method":"EphA3 affinity-isolation from human tissue, EphA3 phosphorylation assays, cell migration and morphology assays, EphA3 siRNA knockdown, in vivo neovascularization assays in immunocompromised mice","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function with in vivo readout and receptor activation assays, single lab","pmids":["25420155"],"is_preprint":false},{"year":2018,"finding":"EphA3 depletion in gastric cancer cells inhibited HUVEC tube formation and migration as well as tumor angiogenesis in vivo, acting through the STAT3/VEGF signaling pathway.","method":"Lentivirus-mediated EphA3 knockdown, tube formation assay, migration assay, xenograft models with microvessel density analysis, STAT3/VEGF pathway western blotting","journal":"Oncology reports","confidence":"Medium","confidence_rationale":"Tier 2-3 — loss-of-function with defined pathway readout and in vivo confirmation, single lab","pmids":["30066881"],"is_preprint":false},{"year":2016,"finding":"EPHA3 overexpression in SCLC cells reduced chemoresistance by increasing apoptosis and G0/G1 arrest with concomitant reduced phosphorylation of the PI3K/BMX/STAT3 signaling pathway; knockdown of EPHA3 promoted resistant phenotype with increased PI3K/BMX/STAT3 phosphorylation, and PI3K or BMX inhibitors reversed this resistance.","method":"EPHA3 overexpression and knockdown in SCLC cells, flow cytometry for apoptosis and cell cycle, pharmacological inhibitors (LY294002, LFM-A13), western blotting of PI3K/BMX/STAT3 pathway, in vivo tumor growth assays","journal":"Tumour biology","confidence":"Medium","confidence_rationale":"Tier 2-3 — gain and loss-of-function with defined pathway readout and inhibitor rescue, single lab","pmids":["27101199"],"is_preprint":false},{"year":2021,"finding":"EphA3 is expressed on CCR10+ mesenchymal progenitor cells in IPF lungs; antibody-mediated (ifabotuzumab) killing of EphA3+ cells significantly reduced CCR10+ cell numbers and ameliorated pulmonary fibrosis in humanized NSG mice.","method":"Human IPF cell characterization, intravenous injection of IPF cells into NSG mice, EphA3-targeted antibody (ifabotuzumab) treatment, histological fibrosis assessment","journal":"JCI insight","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo humanized mouse model with defined cellular and fibrosis readout, single study","pmids":["33945505"],"is_preprint":false},{"year":2013,"finding":"EphA3 knockdown via siRNA screening triggered cellular senescence; selected intracellular EPHA3 tumor-associated point mutations decrease receptor expression level and/or RTK activity, consistent with a tumor suppressor role.","method":"High-content siRNA screen for senescence markers (β-galactosidase, DNA damage, p53/p16 expression), RTK activity assays on EphA3 point mutants","journal":"Cell cycle","confidence":"Medium","confidence_rationale":"Tier 2-3 — siRNA screen with defined senescence readout plus kinase activity assays on mutants, single lab","pmids":["23324396"],"is_preprint":false},{"year":2018,"finding":"EphA3 promotes axon elongation of nasal retinal ganglion cells by competing with EphA4 for ephrin-A binding, thereby reducing ephrin-A-mediated EphA4 forward signaling (decreased EphA4 and ephexin1 phosphorylation) and decreasing axonal filopodia density.","method":"Clustered EphA3-Fc treatment of retinal explants, in vitro and in vivo electroporation with dominant-negative EphA4, immunoprecipitation and western blot for EphA4/ephexin1 phosphorylation, PI-PLC shedding of ephrin-As","journal":"Experimental eye research","confidence":"Medium","confidence_rationale":"Tier 2 — reciprocal approaches (gain and loss-of-function, ligand shedding) with defined phosphorylation and morphological readouts, single lab","pmids":["30237102"],"is_preprint":false},{"year":2021,"finding":"EPHA3 contributes to epigenetic suppression of PTEN in radioresistant head and neck cancer cells via EPHA3/c-Myc signaling that upregulates DNMT1-mediated DNA methylation and EZH2-mediated H3K27me3 histone methylation at the PTEN locus.","method":"Western blotting for DNMT1, EZH2, H3K27me3, and PTEN; EPHA3 manipulation in radioresistant cell lines; correlation of signaling components","journal":"Biomolecules","confidence":"Low","confidence_rationale":"Tier 3 — correlative western blot data without direct ChIP or rescue experiments linking EPHA3 to specific epigenetic writers, single lab","pmids":["33919657"],"is_preprint":false},{"year":2018,"finding":"Androgen receptor (AR) induces EPHA3 expression by forming a transcriptional complex with SP1 that binds SP1 sites (−295 to −261) in the EPHA3 core promoter, as demonstrated by co-immunoprecipitation and chromatin immunoprecipitation assays.","method":"Co-immunoprecipitation, chromatin immunoprecipitation (ChIP), luciferase reporter assays with EPHA3 promoter truncations and SP1 site mutations, siRNA knockdown of AR and SP1","journal":"Oncology reports","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP plus ChIP plus reporter assays with deletion and mutation analysis, single lab","pmids":["29917167"],"is_preprint":false},{"year":2019,"finding":"EPHA3 overexpression in macrophages from endometriosis mice inhibits activation of the mTOR signaling pathway, downregulates bcl-2, upregulates autophagy markers (Atg3, LC3-II/LC3-I, Beclin1) and apoptosis markers (bax, fas), thereby promoting macrophage autophagy and apoptosis.","method":"Mouse endometriosis model, macrophage transfection with EPHA3 vector or siRNA, western blotting for mTOR pathway components and autophagy/apoptosis markers, autophagic lysosome visualization, flow cytometry for apoptosis","journal":"Bioscience reports","confidence":"Low","confidence_rationale":"Tier 3 — gain and loss-of-function with signaling readouts, but no direct mechanistic link between EPHA3 kinase activity and mTOR established, single lab","pmids":["31262977"],"is_preprint":false}],"current_model":"EphA3 is a receptor tyrosine kinase whose kinase activity is autoinhibited by its unphosphorylated juxtamembrane segment (structurally defined by crystallography) and activated upon ephrin-A ligand binding and receptor oligomerization; it can also be silenced in cis by ephrinA5 co-expression independently of the canonical ligand-binding domain, or ligand-independently cleaved by presenilin/γ-secretase to generate an intracellular domain that binds NMIIA to promote axon elongation via RhoA inhibition; in cancer contexts, EphA3 acts as a context-dependent tumor suppressor (suppressing AKT signaling and inducing senescence) or tumor promoter (maintaining GBM tumor-initiating cells via MAPK signaling), with somatic cancer mutations disrupting kinase activity, ligand binding, or receptor surface localization."},"narrative":{"teleology":[{"year":2003,"claim":"Defining how EphA3 assembles signaling-competent oligomeric complexes with ephrin-A5 identified three essential receptor surface areas, one outside previously characterized interaction domains, establishing the structural requirements for ligand recognition and receptor clustering.","evidence":"Random mutagenesis screen of EphA3 in yeast with functional characterization of ephrin-A5 binding and signaling","pmids":["14660665"],"confidence":"High","gaps":["No full-length receptor-ligand co-crystal structure","Contribution of the third novel contact area to in vivo signaling unclear"]},{"year":2006,"claim":"Two parallel discoveries revealed that EphA3 can be silenced in cis by co-expressed ephrinA5 through a mechanism independent of the canonical ligand-binding domain, and that EphA3 is essential for cardiac mesenchymal transformation during septum and cushion development, establishing dual roles in axon guidance regulation and organogenesis.","evidence":"FRET, point mutagenesis, and retinal axon functional assays for cis-silencing; EphA3 knockout mouse analysis with endocardial cushion explant migration assays for cardiac role","pmids":["16491080","17046737"],"confidence":"High","gaps":["Structural basis of the cis interaction is undefined","Which downstream effectors mediate EphA3-driven EMT in the heart is unknown","Whether cis-silencing operates in non-retinal tissues is untested"]},{"year":2008,"claim":"Crystal structures of the EphA3 kinase domain resolved the autoinhibition mechanism: the unphosphorylated juxtamembrane segment directly occludes the active site, and autophosphorylation of two juxtamembrane tyrosines releases catalytic activity through a coupled allosteric pathway connecting the juxtamembrane region, activation loop, and catalytic residues.","evidence":"X-ray crystallography of wild-type and mutant EphA3 kinase domains with and without the juxtamembrane segment, combined with kinase activity assays","pmids":["18547520"],"confidence":"High","gaps":["Structure of full-length EphA3 in an oligomeric signaling complex remains unknown","Role of SAM domain in autoinhibition or activation not addressed"]},{"year":2012,"claim":"Characterization of cancer-associated EphA3 mutations demonstrated that somatic mutations impair kinase activity, ligand binding, or surface expression, and that some act as dominant negatives; wild-type EphA3 re-expression suppressed AKT signaling and tumor growth, establishing EphA3 as an ephrin- and kinase-dependent tumor suppressor disrupted in multiple cancer types.","evidence":"Kinase activity, ephrin-binding, and surface localization assays on mutants; dominant-negative analysis; xenograft tumor growth and apoptosis assays with AKT pathway readout","pmids":["22242939","22829656"],"confidence":"High","gaps":["How dominant-negative mutants suppress wild-type receptor at the structural level is unclear","Patient-level genotype-phenotype correlations are limited"]},{"year":2013,"claim":"EphA3 was shown to have a context-dependent oncogenic role in glioblastoma, where it marks tumor-initiating cells, maintains stemness via MAPK signaling, and its depletion reduces tumorigenic potential—contrasting with its tumor suppressor function in other cancers and complemented by the finding that EphA3 knockdown induces senescence.","evidence":"EphA3 knockdown and EphA3+ cell depletion in GBM with MAPK signaling analysis; siRNA senescence screen with β-galactosidase, DNA damage, and p53/p16 readouts plus kinase assays on tumor-associated mutants","pmids":["23410976","23324396"],"confidence":"High","gaps":["Whether EphA3 kinase activity or scaffolding function drives GBM stemness is unresolved","Direct substrates mediating MAPK-dependent stemness maintenance are unknown"]},{"year":2018,"claim":"A series of studies clarified EphA3's axon guidance mechanism: neurocan blocks NCAM/EphA3 clustering and kinase activation to modulate ephrin-A5-induced growth cone collapse, and EphA3 promotes nasal retinal axon elongation by competing with EphA4 for ephrin-A binding, reducing EphA4/ephexin1 phosphorylation—showing EphA3 acts as both a signal transducer and a competitive inhibitor of co-expressed Eph receptors.","evidence":"NCAM co-immunoprecipitation, EphA3 kinase activation, growth cone collapse assays in cortical interneurons; clustered EphA3-Fc treatment and dominant-negative EphA4 in retinal explants with phosphorylation readouts","pmids":["29670169","30237102"],"confidence":"High","gaps":["Whether EphA3/EphA4 competition operates in non-retinal axon guidance contexts is untested","Stoichiometric requirements for NCAM-EphA3 complex formation are undefined"]},{"year":2019,"claim":"Discovery of ligand-independent presenilin/γ-secretase cleavage of EphA3 revealed a non-canonical signaling mode: the released intracellular domain binds NMIIA, increases Ser1943 phosphorylation to promote filament disassembly, and drives axon elongation via RhoA inhibition, establishing a proteolytic activation pathway distinct from classical ephrin-dependent signaling.","evidence":"PS1/EphA3 KO neurons, proteomic identification of EphA3 ICD interactors, Ser1943 phosphorylation assays, ICD rescue experiments, pharmacological NMII inhibition","pmids":["31577226"],"confidence":"High","gaps":["The kinase or phosphatase directly responsible for NMIIA Ser1943 phosphorylation downstream of EphA3 ICD is unknown","Whether γ-secretase cleavage of EphA3 occurs in non-neuronal tissues is untested","Regulation of the initial extracellular domain shedding step is unclear"]},{"year":2021,"claim":"EphA3 was identified as a therapeutic surface target on mesenchymal progenitor cells in idiopathic pulmonary fibrosis, where antibody-mediated killing of EphA3+ cells reduced fibrosis in a humanized mouse model, extending EphA3's disease relevance beyond cancer.","evidence":"Human IPF cell characterization, ifabotuzumab treatment in humanized NSG mice with fibrosis histological assessment","pmids":["33945505"],"confidence":"Medium","gaps":["Whether EphA3 kinase signaling drives the fibrotic phenotype or merely marks the pathogenic population is unclear","Confirmation in independent cohorts and animal models needed"]},{"year":null,"claim":"Key unresolved questions include the full-length structure of EphA3 in signaling oligomers, the identity of direct EphA3 kinase substrates mediating MAPK- and AKT-dependent cellular outcomes, whether the γ-secretase cleavage pathway operates beyond neurons, and the structural basis by which cis ephrinA5 silences EphA3 signaling.","evidence":"","pmids":[],"confidence":"High","gaps":["No full-length EphA3 signaling complex structure","Direct kinase substrates largely unidentified","In vivo relevance of γ-secretase cleavage in non-neuronal tissues untested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,5,6,13]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,14]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[2,5,8]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,4,6,7,11]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[3,14]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[4,5,6,13]}],"complexes":[],"partners":["EFNA5","EFNA1","NCAM1","MYH9","PSEN1","EPHA4"],"other_free_text":[]},"mechanistic_narrative":"EphA3 is a receptor tyrosine kinase that transduces ephrin-A ligand signals to control cell migration, axon guidance, and tissue morphogenesis during development and in disease. Its kinase domain is autoinhibited by an unphosphorylated juxtamembrane segment that blocks the active site; autophosphorylation of two juxtamembrane tyrosines relieves this inhibition and enables signal transduction through downstream pathways including MAPK, AKT, and PI3K/BMX/STAT3 [PMID:18547520, PMID:22829656, PMID:23410976]. In the nervous system, EphA3 modulates axon elongation both through classical ephrin-A-dependent forward signaling and through ligand-independent presenilin/γ-secretase cleavage that generates an intracellular domain binding non-muscle myosin IIA to inhibit RhoA and promote cytoskeletal rearrangement [PMID:31577226, PMID:30237102]; cis-interaction with ephrinA5 silences EphA3 trans-signaling independently of its canonical ligand-binding domain [PMID:16491080]. In cancer, EphA3 functions as a context-dependent tumor suppressor—somatic mutations disrupt kinase activity, ligand binding, or surface localization, and wild-type re-expression suppresses AKT signaling and tumor growth—while in glioblastoma it maintains tumor-initiating cells via MAPK signaling [PMID:22242939, PMID:22829656, PMID:23410976]."},"prefetch_data":{"uniprot":{"accession":"P29320","full_name":"Ephrin type-A receptor 3","aliases":["EPH-like kinase 4","EK4","hEK4","HEK","Human embryo kinase","Tyrosine-protein kinase TYRO4","Tyrosine-protein kinase receptor ETK1","Eph-like tyrosine kinase 1"],"length_aa":983,"mass_kda":110.1,"function":"Receptor tyrosine kinase which binds promiscuously membrane-bound ephrin family ligands residing on adjacent cells, leading to contact-dependent bidirectional signaling into neighboring cells. The signaling pathway downstream of the receptor is referred to as forward signaling while the signaling pathway downstream of the ephrin ligand is referred to as reverse signaling (PubMed:11870224, PubMed:12794130). Highly promiscuous for ephrin-A ligands it binds preferentially EFNA5 (By similarity). Upon activation by EFNA5 regulates cell-cell adhesion, cytoskeletal organization and cell migration (PubMed:11870224). Also activated by EFNA1, inhibiting epithelial-to-mesenchymal transition of cardiac cells and playing a role in heart development (PubMed:12794130). Involved in the retinotectal mapping of neurons. May also control the segregation but not the guidance of motor and sensory axons during neuromuscular circuit development (By similarity)","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/P29320/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EPHA3","classification":"Not Classified","n_dependent_lines":5,"n_total_lines":1208,"dependency_fraction":0.0041390728476821195},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/EPHA3","total_profiled":1310},"omim":[{"mim_id":"610328","title":"RUN AND FYVE DOMAINS-CONTAINING PROTEIN 2; RUFY2","url":"https://www.omim.org/entry/610328"},{"mim_id":"610327","title":"RUN AND FYVE DOMAINS-CONTAINING PROTEIN 1; RUFY1","url":"https://www.omim.org/entry/610327"},{"mim_id":"608892","title":"CHROMODOMAIN HELICASE DNA-BINDING PROTEIN 7; CHD7","url":"https://www.omim.org/entry/608892"},{"mim_id":"602192","title":"A DISINTEGRIN AND METALLOPROTEINASE DOMAIN 10; ADAM10","url":"https://www.omim.org/entry/602192"},{"mim_id":"602190","title":"EPHRIN RECEPTOR EphA7; EPHA7","url":"https://www.omim.org/entry/602190"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nuclear membrane","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Supported"},{"location":"Cytosol","reliability":"Supported"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Actin filaments","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"prostate","ntpm":29.5}],"url":"https://www.proteinatlas.org/search/EPHA3"},"hgnc":{"alias_symbol":["HEK","HEK4"],"prev_symbol":["ETK","ETK1","TYRO4"]},"alphafold":{"accession":"P29320","domains":[{"cath_id":"2.60.120.260","chopping":"28-199","consensus_level":"high","plddt":88.7751,"start":28,"end":199},{"cath_id":"2.60.40.1770","chopping":"204-259","consensus_level":"medium","plddt":90.5427,"start":204,"end":259},{"cath_id":"2.60.40.10","chopping":"329-430","consensus_level":"high","plddt":91.0727,"start":329,"end":430},{"cath_id":"2.60.40.10","chopping":"442-527","consensus_level":"high","plddt":90.0335,"start":442,"end":527},{"cath_id":"3.30.200.20","chopping":"598-699","consensus_level":"high","plddt":84.7519,"start":598,"end":699},{"cath_id":"1.10.510.10","chopping":"706-887","consensus_level":"high","plddt":86.9299,"start":706,"end":887},{"cath_id":"1.10.150.50","chopping":"915-976","consensus_level":"medium","plddt":52.0382,"start":915,"end":976}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P29320","model_url":"https://alphafold.ebi.ac.uk/files/AF-P29320-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P29320-F1-predicted_aligned_error_v6.png","plddt_mean":80.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EPHA3","jax_strain_url":"https://www.jax.org/strain/search?query=EPHA3"},"sequence":{"accession":"P29320","fasta_url":"https://rest.uniprot.org/uniprotkb/P29320.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P29320/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P29320"}},"corpus_meta":[{"pmid":"23410976","id":"PMC_23410976","title":"EphA3 maintains tumorigenicity and is a therapeutic target in glioblastoma multiforme.","date":"2013","source":"Cancer cell","url":"https://pubmed.ncbi.nlm.nih.gov/23410976","citation_count":175,"is_preprint":false},{"pmid":"11713275","id":"PMC_11713275","title":"Neuropeptide-induced androgen independence in prostate cancer cells: roles of nonreceptor tyrosine kinases Etk/Bmx, Src, and focal adhesion kinase.","date":"2001","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/11713275","citation_count":157,"is_preprint":false},{"pmid":"16491080","id":"PMC_16491080","title":"Silencing of EphA3 through a cis interaction with ephrinA5.","date":"2006","source":"Nature neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/16491080","citation_count":154,"is_preprint":false},{"pmid":"11331870","id":"PMC_11331870","title":"Regulation of the PH-domain-containing tyrosine kinase Etk by focal adhesion kinase through the FERM domain.","date":"2001","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/11331870","citation_count":121,"is_preprint":false},{"pmid":"10688651","id":"PMC_10688651","title":"Etk, a Btk family tyrosine kinase, mediates cellular transformation by linking Src to STAT3 activation.","date":"2000","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/10688651","citation_count":115,"is_preprint":false},{"pmid":"9556622","id":"PMC_9556622","title":"Comparison of Na-K-Cl cotransporters. NKCC1, NKCC2, and the HEK cell Na-L-Cl cotransporter.","date":"1998","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9556622","citation_count":110,"is_preprint":false},{"pmid":"11382770","id":"PMC_11382770","title":"Etk/Bmx tyrosine kinase activates Pak1 and regulates tumorigenicity of breast cancer cells.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11382770","citation_count":97,"is_preprint":false},{"pmid":"30176744","id":"PMC_30176744","title":"Nose-to-brain delivery of temozolomide-loaded PLGA nanoparticles functionalized with anti-EPHA3 for glioblastoma targeting.","date":"2018","source":"Drug delivery","url":"https://pubmed.ncbi.nlm.nih.gov/30176744","citation_count":92,"is_preprint":false},{"pmid":"7838529","id":"PMC_7838529","title":"Ligands for the receptor tyrosine kinases hek and elk: isolation of cDNAs encoding a family of proteins.","date":"1995","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/7838529","citation_count":83,"is_preprint":false},{"pmid":"14660665","id":"PMC_14660665","title":"Dissecting the EphA3/Ephrin-A5 interactions using a novel functional mutagenesis screen.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14660665","citation_count":77,"is_preprint":false},{"pmid":"16932810","id":"PMC_16932810","title":"Critical function of Bmx/Etk in ischemia-mediated arteriogenesis and angiogenesis.","date":"2006","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/16932810","citation_count":75,"is_preprint":false},{"pmid":"10362360","id":"PMC_10362360","title":"Etk/Bmx, a PH-domain containing tyrosine kinase, protects prostate cancer cells from apoptosis induced by photodynamic therapy or thapsigargin.","date":"1999","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/10362360","citation_count":66,"is_preprint":false},{"pmid":"15569306","id":"PMC_15569306","title":"D1 dopamine receptor signaling involves caveolin-2 in HEK-293 cells.","date":"2004","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/15569306","citation_count":66,"is_preprint":false},{"pmid":"29208667","id":"PMC_29208667","title":"Etk Interaction with PFKFB4 Modulates Chemoresistance of Small-cell Lung Cancer by Regulating Autophagy.","date":"2017","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/29208667","citation_count":65,"is_preprint":false},{"pmid":"8397371","id":"PMC_8397371","title":"PCR mediated detection of a new human receptor-tyrosine-kinase, HEK 2.","date":"1993","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/8397371","citation_count":64,"is_preprint":false},{"pmid":"16941478","id":"PMC_16941478","title":"Somatic mutations of GUCY2F, EPHA3, and NTRK3 in human cancers.","date":"2006","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/16941478","citation_count":63,"is_preprint":false},{"pmid":"11060230","id":"PMC_11060230","title":"Two homeobox genes define the domain of EphA3 expression in the developing chick retina.","date":"2000","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/11060230","citation_count":63,"is_preprint":false},{"pmid":"17046737","id":"PMC_17046737","title":"A critical role for the EphA3 receptor tyrosine kinase in heart development.","date":"2006","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/17046737","citation_count":60,"is_preprint":false},{"pmid":"18497741","id":"PMC_18497741","title":"Structure of Escherichia coli tyrosine kinase Etk reveals a novel activation mechanism.","date":"2008","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/18497741","citation_count":60,"is_preprint":false},{"pmid":"21354312","id":"PMC_21354312","title":"Design of Experiment in CHO and HEK transient transfection condition optimization.","date":"2011","source":"Protein expression and purification","url":"https://pubmed.ncbi.nlm.nih.gov/21354312","citation_count":60,"is_preprint":false},{"pmid":"12970074","id":"PMC_12970074","title":"Protein kinase C-mediated Ca2+ entry in HEK 293 cells transiently expressing human TRPV4.","date":"2003","source":"British journal of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/12970074","citation_count":59,"is_preprint":false},{"pmid":"33417456","id":"PMC_33417456","title":"Intranasal Delivery of Temozolomide-Conjugated Gold Nanoparticles Functionalized with Anti-EphA3 for Glioblastoma Targeting.","date":"2021","source":"Molecular pharmaceutics","url":"https://pubmed.ncbi.nlm.nih.gov/33417456","citation_count":56,"is_preprint":false},{"pmid":"20570899","id":"PMC_20570899","title":"Compensatory upregulation of tyrosine kinase Etk/BMX in response to androgen deprivation promotes castration-resistant growth of prostate cancer cells.","date":"2010","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/20570899","citation_count":56,"is_preprint":false},{"pmid":"12483534","id":"PMC_12483534","title":"Coordinating Etk/Bmx activation and VEGF upregulation to promote cell survival and proliferation.","date":"2002","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/12483534","citation_count":56,"is_preprint":false},{"pmid":"22242939","id":"PMC_22242939","title":"Cancer somatic mutations disrupt functions of the EphA3 receptor tyrosine kinase through multiple mechanisms.","date":"2012","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22242939","citation_count":54,"is_preprint":false},{"pmid":"22829656","id":"PMC_22829656","title":"Effects of cancer-associated EPHA3 mutations on lung cancer.","date":"2012","source":"Journal of the National Cancer Institute","url":"https://pubmed.ncbi.nlm.nih.gov/22829656","citation_count":53,"is_preprint":false},{"pmid":"18547520","id":"PMC_18547520","title":"Autoregulation by the juxtamembrane region of the human ephrin receptor tyrosine kinase A3 (EphA3).","date":"2008","source":"Structure (London, England : 1993)","url":"https://pubmed.ncbi.nlm.nih.gov/18547520","citation_count":53,"is_preprint":false},{"pmid":"25391995","id":"PMC_25391995","title":"EphA3 biology and cancer.","date":"2014","source":"Growth factors (Chur, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/25391995","citation_count":52,"is_preprint":false},{"pmid":"12023958","id":"PMC_12023958","title":"Selective activation of small GTPase RhoA by tyrosine kinase Etk through its pleckstrin homology domain.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12023958","citation_count":52,"is_preprint":false},{"pmid":"24696141","id":"PMC_24696141","title":"PACAP-induced ERK activation in HEK cells expressing PAC1 receptors involves both receptor internalization and PKC signaling.","date":"2014","source":"American journal of physiology. Cell physiology","url":"https://pubmed.ncbi.nlm.nih.gov/24696141","citation_count":47,"is_preprint":false},{"pmid":"13129920","id":"PMC_13129920","title":"Expression of Drosophila trehalose-phosphate synthase in HEK-293 cells increases hypoxia tolerance.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/13129920","citation_count":47,"is_preprint":false},{"pmid":"24943513","id":"PMC_24943513","title":"Introducing PeptoPlexes: polylysine-block-polysarcosine based polyplexes for transfection of HEK 293T cells.","date":"2014","source":"Macromolecular bioscience","url":"https://pubmed.ncbi.nlm.nih.gov/24943513","citation_count":45,"is_preprint":false},{"pmid":"15979128","id":"PMC_15979128","title":"Endogenous channels in HEK cells and potential roles in HCN ionic current measurements.","date":"2005","source":"Progress in biophysics and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/15979128","citation_count":45,"is_preprint":false},{"pmid":"22350700","id":"PMC_22350700","title":"Aberrant expression of EphA3 in gastric carcinoma: correlation with tumor angiogenesis and survival.","date":"2012","source":"Journal of gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/22350700","citation_count":44,"is_preprint":false},{"pmid":"16912182","id":"PMC_16912182","title":"Tyrosine kinase Etk/BMX is up-regulated in human prostate cancer and its overexpression induces prostate intraepithelial neoplasia in mouse.","date":"2006","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/16912182","citation_count":44,"is_preprint":false},{"pmid":"29888905","id":"PMC_29888905","title":"N-Glycosylation of Extracellular Vesicles from HEK-293 and Glioma Cell Lines.","date":"2018","source":"Analytical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29888905","citation_count":42,"is_preprint":false},{"pmid":"25847125","id":"PMC_25847125","title":"DNA damage and S phase arrest induced by Ochratoxin A in human embryonic kidney cells (HEK 293).","date":"2014","source":"Mutation research","url":"https://pubmed.ncbi.nlm.nih.gov/25847125","citation_count":42,"is_preprint":false},{"pmid":"15504339","id":"PMC_15504339","title":"Silencing of SARS-CoV spike gene by small interfering RNA in HEK 293T cells.","date":"2004","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/15504339","citation_count":41,"is_preprint":false},{"pmid":"16276002","id":"PMC_16276002","title":"Human single-domain neutralizing intrabodies directed against Etk kinase: a novel approach to impair cellular transformation.","date":"2005","source":"Molecular cancer therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/16276002","citation_count":40,"is_preprint":false},{"pmid":"16083359","id":"PMC_16083359","title":"Upregulation of EphA3 receptor after spinal cord injury.","date":"2005","source":"Journal of neurotrauma","url":"https://pubmed.ncbi.nlm.nih.gov/16083359","citation_count":38,"is_preprint":false},{"pmid":"25188519","id":"PMC_25188519","title":"Targeting Btk/Etk of prostate cancer cells by a novel dual inhibitor.","date":"2014","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/25188519","citation_count":37,"is_preprint":false},{"pmid":"27101199","id":"PMC_27101199","title":"EPHA3 regulates the multidrug resistance of small cell lung cancer via the PI3K/BMX/STAT3 signaling pathway.","date":"2016","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/27101199","citation_count":37,"is_preprint":false},{"pmid":"14726663","id":"PMC_14726663","title":"The Tyr-kinase inhibitor AG879, that blocks the ETK-PAK1 interaction, suppresses the RAS-induced PAK1 activation and malignant transformation.","date":"2004","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/14726663","citation_count":35,"is_preprint":false},{"pmid":"31778795","id":"PMC_31778795","title":"Functional characterization of odorant receptors from the moth Eriocrania semipurpurella: A comparison of results in the Xenopus oocyte and HEK cell systems.","date":"2019","source":"Insect biochemistry and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/31778795","citation_count":35,"is_preprint":false},{"pmid":"21464992","id":"PMC_21464992","title":"Phosphorylation and transport in the Na-K-2Cl cotransporters, NKCC1 and NKCC2A, compared in HEK-293 cells.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21464992","citation_count":35,"is_preprint":false},{"pmid":"14676838","id":"PMC_14676838","title":"The role of tyrosine kinase Etk/Bmx in EGF-induced apoptosis of MDA-MB-468 breast cancer cells.","date":"2004","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/14676838","citation_count":32,"is_preprint":false},{"pmid":"22675501","id":"PMC_22675501","title":"Cycling of Etk and Etp phosphorylation states is involved in formation of group 4 capsule by Escherichia coli.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22675501","citation_count":31,"is_preprint":false},{"pmid":"32621117","id":"PMC_32621117","title":"Critical role of EphA3 in cancer and current state of EphA3 drug therapeutics.","date":"2020","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/32621117","citation_count":30,"is_preprint":false},{"pmid":"29670169","id":"PMC_29670169","title":"Perineuronal Net Protein Neurocan Inhibits NCAM/EphA3 Repellent Signaling in GABAergic Interneurons.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29670169","citation_count":30,"is_preprint":false},{"pmid":"17241243","id":"PMC_17241243","title":"The Hek outer membrane protein of Escherichia coli is an auto-aggregating adhesin and invasin.","date":"2007","source":"FEMS microbiology letters","url":"https://pubmed.ncbi.nlm.nih.gov/17241243","citation_count":30,"is_preprint":false},{"pmid":"16814990","id":"PMC_16814990","title":"A novel role for protein-tyrosine kinase Etk from Escherichia coli K-12 related to polymyxin resistance.","date":"2006","source":"Research in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/16814990","citation_count":29,"is_preprint":false},{"pmid":"21454190","id":"PMC_21454190","title":"Copy number variations of EphA3 are associated with multiple types of hematologic malignancies.","date":"2011","source":"Clinical lymphoma, myeloma & leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/21454190","citation_count":29,"is_preprint":false},{"pmid":"28514765","id":"PMC_28514765","title":"BMX/Etk promotes cell proliferation and tumorigenicity of cervical cancer cells through PI3K/AKT/mTOR and STAT3 pathways.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/28514765","citation_count":29,"is_preprint":false},{"pmid":"28198671","id":"PMC_28198671","title":"Transcriptome analysis reveals dynamic changes in coxsackievirus A16 infected HEK 293T cells.","date":"2017","source":"BMC genomics","url":"https://pubmed.ncbi.nlm.nih.gov/28198671","citation_count":28,"is_preprint":false},{"pmid":"28401017","id":"PMC_28401017","title":"miR-495 promotes the chemoresistance of SCLC through the epithelial-mesenchymal transition via Etk/BMX.","date":"2017","source":"American journal of cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/28401017","citation_count":27,"is_preprint":false},{"pmid":"11278797","id":"PMC_11278797","title":"Proteolytic activation of ETK/Bmx tyrosine kinase by caspases.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11278797","citation_count":27,"is_preprint":false},{"pmid":"20559570","id":"PMC_20559570","title":"Etk/Bmx regulates proteinase-activated-receptor1 (PAR1) in breast cancer invasion: signaling partners, hierarchy and physiological significance.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/20559570","citation_count":26,"is_preprint":false},{"pmid":"21920850","id":"PMC_21920850","title":"Mechanisms of melanocortin-2 receptor (MC2R) internalization and recycling in human embryonic kidney (hek) cells: identification of Key Ser/Thr (S/T) amino acids.","date":"2011","source":"Molecular endocrinology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/21920850","citation_count":24,"is_preprint":false},{"pmid":"27313717","id":"PMC_27313717","title":"Chelerythrine chloride induces apoptosis in renal cancer HEK-293 and SW-839 cell lines.","date":"2016","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/27313717","citation_count":23,"is_preprint":false},{"pmid":"25683738","id":"PMC_25683738","title":"Transcriptional and post-transcriptional limitations of high-yielding, PEI-mediated transient transfection with CHO and HEK-293E cells.","date":"2015","source":"Biotechnology progress","url":"https://pubmed.ncbi.nlm.nih.gov/25683738","citation_count":22,"is_preprint":false},{"pmid":"22127760","id":"PMC_22127760","title":"Reduced glutamine concentration improves protein production in growth-arrested CHO-DG44 and HEK-293E cells.","date":"2011","source":"Biotechnology letters","url":"https://pubmed.ncbi.nlm.nih.gov/22127760","citation_count":22,"is_preprint":false},{"pmid":"23321269","id":"PMC_23321269","title":"Silencing of the WFS1 gene in HEK cells induces pathways related to neurodegeneration and mitochondrial damage.","date":"2013","source":"Physiological genomics","url":"https://pubmed.ncbi.nlm.nih.gov/23321269","citation_count":22,"is_preprint":false},{"pmid":"24606948","id":"PMC_24606948","title":"The expression and role of tyrosine kinase ETK/BMX in renal cell carcinoma.","date":"2014","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/24606948","citation_count":21,"is_preprint":false},{"pmid":"19428708","id":"PMC_19428708","title":"Identification and expression analysis of miRNAs during batch culture of HEK-293 cells.","date":"2009","source":"Journal of biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/19428708","citation_count":21,"is_preprint":false},{"pmid":"35774066","id":"PMC_35774066","title":"Production of Lentiviral Vectors Using a HEK-293 Producer Cell Line and Advanced Perfusion Processing.","date":"2022","source":"Frontiers in bioengineering and biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/35774066","citation_count":20,"is_preprint":false},{"pmid":"20004564","id":"PMC_20004564","title":"Non-receptor tyrosine kinase Etk regulation of drug resistance in small-cell lung cancer.","date":"2009","source":"European journal of cancer (Oxford, England : 1990)","url":"https://pubmed.ncbi.nlm.nih.gov/20004564","citation_count":20,"is_preprint":false},{"pmid":"36742140","id":"PMC_36742140","title":"Effect of trans-sodium crocetinate on contrast-induced cytotoxicity in HEK-293 cells.","date":"2023","source":"Iranian journal of basic medical sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36742140","citation_count":20,"is_preprint":false},{"pmid":"20592104","id":"PMC_20592104","title":"Biotin regulates the expression of holocarboxylase synthetase in the miR-539 pathway in HEK-293 cells.","date":"2010","source":"The Journal of nutrition","url":"https://pubmed.ncbi.nlm.nih.gov/20592104","citation_count":20,"is_preprint":false},{"pmid":"17097906","id":"PMC_17097906","title":"Transcriptional profiling of batch and fed-batch protein-free 293-HEK cultures.","date":"2006","source":"Metabolic engineering","url":"https://pubmed.ncbi.nlm.nih.gov/17097906","citation_count":20,"is_preprint":false},{"pmid":"21190559","id":"PMC_21190559","title":"Sperry versus Hebb: topographic mapping in Isl2/EphA3 mutant mice.","date":"2010","source":"BMC neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/21190559","citation_count":20,"is_preprint":false},{"pmid":"19342512","id":"PMC_19342512","title":"Cross-inhibition between nicotinic acetylcholine receptors and P2X receptors in myenteric neurons and HEK-293 cells.","date":"2009","source":"American journal of physiology. Gastrointestinal and liver physiology","url":"https://pubmed.ncbi.nlm.nih.gov/19342512","citation_count":20,"is_preprint":false},{"pmid":"33112237","id":"PMC_33112237","title":"Probing the ionotropic activity of glutamate GluD2 receptor in HEK cells with genetically-engineered photopharmacology.","date":"2020","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/33112237","citation_count":20,"is_preprint":false},{"pmid":"27292079","id":"PMC_27292079","title":"Luteolin inhibits GABAA receptors in HEK cells and brain slices.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27292079","citation_count":19,"is_preprint":false},{"pmid":"30346389","id":"PMC_30346389","title":"Rapid Isolation of the Mitoribosome from HEK Cells.","date":"2018","source":"Journal of visualized experiments : JoVE","url":"https://pubmed.ncbi.nlm.nih.gov/30346389","citation_count":19,"is_preprint":false},{"pmid":"37435968","id":"PMC_37435968","title":"Peptide ligands for the affinity purification of adeno-associated viruses from HEK 293 cell lysates.","date":"2023","source":"Biotechnology and bioengineering","url":"https://pubmed.ncbi.nlm.nih.gov/37435968","citation_count":19,"is_preprint":false},{"pmid":"31262977","id":"PMC_31262977","title":"EPHA3 enhances macrophage autophagy and apoptosis by disrupting the mTOR signaling pathway in mice with endometriosis.","date":"2019","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/31262977","citation_count":18,"is_preprint":false},{"pmid":"33945505","id":"PMC_33945505","title":"Antibody-mediated depletion of CCR10+EphA3+ cells ameliorates fibrosis in IPF.","date":"2021","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/33945505","citation_count":18,"is_preprint":false},{"pmid":"31821898","id":"PMC_31821898","title":"Mitochondrial mRNA fragments are circularized in a human HEK cell line.","date":"2019","source":"Mitochondrion","url":"https://pubmed.ncbi.nlm.nih.gov/31821898","citation_count":18,"is_preprint":false},{"pmid":"37445652","id":"PMC_37445652","title":"LRRK2 Knockout Confers Resistance in HEK-293 Cells to Rotenone-Induced Oxidative Stress, Mitochondrial Damage, and Apoptosis.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/37445652","citation_count":18,"is_preprint":false},{"pmid":"30066881","id":"PMC_30066881","title":"EphA3 contributes to tumor growth and angiogenesis in human gastric cancer cells.","date":"2018","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/30066881","citation_count":17,"is_preprint":false},{"pmid":"23324396","id":"PMC_23324396","title":"A high-content cellular senescence screen identifies candidate tumor suppressors, including EPHA3.","date":"2013","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/23324396","citation_count":17,"is_preprint":false},{"pmid":"29660208","id":"PMC_29660208","title":"MiR-340 suppresses the metastasis by targeting EphA3 in cervical cancer.","date":"2018","source":"Cell biology international","url":"https://pubmed.ncbi.nlm.nih.gov/29660208","citation_count":16,"is_preprint":false},{"pmid":"39111833","id":"PMC_39111833","title":"EphA3-targeted chimeric antigen receptor T cells are effective in glioma and generate curative memory T cell responses.","date":"2024","source":"Journal for immunotherapy of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/39111833","citation_count":16,"is_preprint":false},{"pmid":"25978371","id":"PMC_25978371","title":"Antitumor Effects of Oncolytic Adenovirus-Carrying siRNA Targeting Potential Oncogene EphA3.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25978371","citation_count":15,"is_preprint":false},{"pmid":"34446852","id":"PMC_34446852","title":"Signaling profiles in HEK 293T cells co-expressing GLP-1 and GIP receptors.","date":"2021","source":"Acta pharmacologica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/34446852","citation_count":15,"is_preprint":false},{"pmid":"31577226","id":"PMC_31577226","title":"Presenilin/γ-secretase-dependent EphA3 processing mediates axon elongation through non-muscle myosin IIA.","date":"2019","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/31577226","citation_count":14,"is_preprint":false},{"pmid":"21339702","id":"PMC_21339702","title":"Tyrosine kinase Etk/BMX protects nasopharyngeal carcinoma cells from apoptosis induced by radiation.","date":"2011","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/21339702","citation_count":14,"is_preprint":false},{"pmid":"33919657","id":"PMC_33919657","title":"EPHA3 Contributes to Epigenetic Suppression of PTEN in Radioresistant Head and Neck Cancer.","date":"2021","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/33919657","citation_count":14,"is_preprint":false},{"pmid":"25420155","id":"PMC_25420155","title":"Hypoxia-controlled EphA3 marks a human endometrium-derived multipotent mesenchymal stromal cell that supports vascular growth.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25420155","citation_count":14,"is_preprint":false},{"pmid":"18674612","id":"PMC_18674612","title":"TRP-2 expression protects HEK cells from dopamine- and hydroquinone-induced toxicity.","date":"2008","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/18674612","citation_count":14,"is_preprint":false},{"pmid":"30483759","id":"PMC_30483759","title":"EphA3 inhibits migration and invasion of esophageal cancer cells by activating the mesenchymal‑epithelial transition process.","date":"2018","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/30483759","citation_count":13,"is_preprint":false},{"pmid":"33493920","id":"PMC_33493920","title":"Designing and evaluation of MERS-CoV siRNAs in HEK-293 cell line.","date":"2020","source":"Journal of infection and public health","url":"https://pubmed.ncbi.nlm.nih.gov/33493920","citation_count":13,"is_preprint":false},{"pmid":"36449077","id":"PMC_36449077","title":"Apolipoprotein L1 (APOL1) cation current in HEK-293 cells and in human podocytes.","date":"2022","source":"Pflugers Archiv : European journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/36449077","citation_count":13,"is_preprint":false},{"pmid":"19820835","id":"PMC_19820835","title":"Infection kinetics of human adenovirus serotype 41 in HEK 293 cells.","date":"2009","source":"Memorias do Instituto Oswaldo Cruz","url":"https://pubmed.ncbi.nlm.nih.gov/19820835","citation_count":13,"is_preprint":false},{"pmid":"30237102","id":"PMC_30237102","title":"Regulation of axonal EphA4 forward signaling is involved in the effect of EphA3 on chicken retinal ganglion cell axon growth during retinotectal mapping.","date":"2018","source":"Experimental eye research","url":"https://pubmed.ncbi.nlm.nih.gov/30237102","citation_count":13,"is_preprint":false},{"pmid":"17479408","id":"PMC_17479408","title":"JWA gene is involved in cadmium-induced growth inhibition and apoptosis in HEK-293T cells.","date":"2007","source":"Journal of toxicology and environmental health. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/17479408","citation_count":13,"is_preprint":false},{"pmid":"10545030","id":"PMC_10545030","title":"Regulatory interrelations among topographic molecules CBF1, CBF2 and EphA3 in the developing chick retina.","date":"1999","source":"Development, growth & differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/10545030","citation_count":13,"is_preprint":false},{"pmid":"29917167","id":"PMC_29917167","title":"Androgen receptor induces EPHA3 expression by interacting with transcription factor SP1.","date":"2018","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/29917167","citation_count":12,"is_preprint":false},{"pmid":"21864196","id":"PMC_21864196","title":"Reporter gene HEK 293 cells and WNT/Frizzled fusion proteins as tools to study WNT signaling pathways.","date":"2011","source":"Biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21864196","citation_count":12,"is_preprint":false},{"pmid":"17058277","id":"PMC_17058277","title":"A gas-inducible expression system in HEK.EBNA cells applied to controlled proliferation studies by expression of p27(Kip1).","date":"2007","source":"Biotechnology and bioengineering","url":"https://pubmed.ncbi.nlm.nih.gov/17058277","citation_count":12,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":51263,"output_tokens":4454,"usd":0.110299},"stage2":{"model":"claude-opus-4-6","input_tokens":7926,"output_tokens":2757,"usd":0.162832},"total_usd":0.273131,"stage1_batch_id":"msgbatch_0118RScg3WGaGkaZRtWZbtce","stage2_batch_id":"msgbatch_01Cpfzvtbj42aqp5oYZ5Te2s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2006,\n      \"finding\": \"EphA3 undergoes a cis interaction with ephrinA5 that is independent of the established ligand-binding domain of EphA3; this cis interaction abolishes tyrosine phosphorylation of EphA3 and results in loss of sensitivity of retinal axons to ephrinAs in trans, effectively silencing EphA3 signaling.\",\n      \"method\": \"Truncated EphA3 constructs, single-amino acid point mutants of ephrinA5, fluorescence resonance energy transfer (FRET), and functional retinal axon assays\",\n      \"journal\": \"Nature neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal methods (FRET, point mutagenesis, functional axon assays) in a single rigorous study\",\n      \"pmids\": [\"16491080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"The crystal structure of the EphA3 kinase domain revealed that the unphosphorylated juxtamembrane region blocks catalysis by interacting with the active site; autophosphorylation of two juxtamembrane tyrosines relieves this inhibition, and a conserved coupled pathway of residues connects the juxtamembrane segment, activation loop, and catalytic residues.\",\n      \"method\": \"High-resolution X-ray crystallography of wild-type and site-specific mutants of EphA3 kinase domain with and without juxtamembrane segment; kinase activity assays\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structures of multiple states combined with mutagenesis and kinase activity assays\",\n      \"pmids\": [\"18547520\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Three EphA3 surface areas are essential for EphA3/ephrin-A5 recognition and assembly of signaling-competent oligomeric receptor-ligand complexes; two map to previously identified Eph/ephrin interfaces and a third is a novel contact area outside the structurally characterized interaction domains.\",\n      \"method\": \"Random mutagenesis of EphA3 expressed in yeast, selection for impaired ephrin-A5 binding, functional characterization of point mutants\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — functional mutagenesis screen with rigorous characterization of binding and signaling\",\n      \"pmids\": [\"14660665\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"EphA3 null mice exhibit significant defects in atrial septa and atrioventricular endocardial cushion development; EphA3 and its ligand ephrin-A1 are expressed in adjacent cells in the developing endocardial cushions, and EphA3-/- endocardial cushion explants produce fewer migrating mesenchymal cells, indicating EphA3 signaling drives epithelial-to-mesenchymal transformation during heart development.\",\n      \"method\": \"EphA3 knockout mouse phenotypic analysis, endocardial cushion explant migration assays, expression analysis of EphA3 and ephrin-A1\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with defined cellular phenotype (EMT defect) and mechanistic localization of ligand-receptor pair\",\n      \"pmids\": [\"17046737\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EphA3 is highly expressed on tumor-initiating cells in glioblastoma and maintains cells in a less differentiated state by modulating MAPK signaling; EphA3 knockdown or depletion of EphA3-positive cells reduced tumorigenic potential.\",\n      \"method\": \"EphA3 knockdown, depletion of EphA3-positive tumor cells, radiolabelled EphA3-specific monoclonal antibody treatment, MAPK signaling analysis in GBM\",\n      \"journal\": \"Cancer cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with specific tumorigenic and signaling readouts, multiple approaches\",\n      \"pmids\": [\"23410976\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Multiple EphA3 somatic mutations found in lung, colorectal, and hepatocellular cancers impair kinase activity, ephrin ligand binding, and/or decrease receptor cell surface localization, suggesting EphA3 has ephrin- and kinase-dependent tumor-suppressing activities disrupted by cancer mutations.\",\n      \"method\": \"Kinase activity assays, ephrin binding assays, cell surface localization analysis of mutant receptors\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal biochemical assays on cancer-associated mutants\",\n      \"pmids\": [\"22242939\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"At least two cancer-associated EPHA3 somatic mutations function as dominant inhibitors of wild-type EPHA3 protein; wild-type EPHA3 re-expression in lung cancer lines increased apoptosis through suppression of AKT activation and inhibited tumor xenograft growth.\",\n      \"method\": \"Immunoprecipitation, western blotting, kinase assays, dominant-negative analysis, xenograft tumor growth assays, apoptosis assays\",\n      \"journal\": \"Journal of the National Cancer Institute\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro kinase assays and in vivo xenograft experiments with mechanistic AKT pathway readout\",\n      \"pmids\": [\"22829656\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Presenilin-1/γ-secretase cleaves EphA3 independently of ephrin ligand binding, generating an intracellular domain (ICD) fragment; EphA3 ICD binds non-muscle myosin IIA (NMIIA) and increases its phosphorylation at Ser1943, promoting NMIIA filament disassembly and cytoskeleton rearrangement, thereby driving axon elongation through inhibition of RhoA signaling.\",\n      \"method\": \"Genetic PS1/EphA3 knockout neurons, proteomic analysis of EphA3 ICD interactors, phosphorylation assays, rescue experiments with ICD fragment, pharmacological NMII inhibition\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal methods including proteomics, KO genetics, rescue experiments, and phosphorylation assays\",\n      \"pmids\": [\"31577226\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Neurocan (a perineuronal net component) binds NCAM at the EphA3-binding site within the immunoglobulin-2 domain, inhibiting NCAM/EphA3 association, membrane clustering of NCAM/EphA3 in cortical interneuron axons, EphA3 kinase activation, and ephrin-A5-induced growth cone collapse.\",\n      \"method\": \"Binding assays, co-immunoprecipitation, EphA3 kinase activation assays, growth cone collapse assays in cortical interneuron cultures, organotypic cortical slice cultures with enzymatic PNN removal\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods including binding assays, kinase assays, and functional growth cone assays\",\n      \"pmids\": [\"29670169\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Hypoxia-inducible factor (HIF)-1α controls EphA3 expression in human endometrial multipotent mesenchymal stromal cells (eMSCs); EphA3 agonists induce receptor phosphorylation, cell contraction, cell-cell segregation and directed cell migration, and EphA3 silencing significantly inhibits transplanted eMSC support of neovascularisation in vivo.\",\n      \"method\": \"EphA3 affinity-isolation from human tissue, EphA3 phosphorylation assays, cell migration and morphology assays, EphA3 siRNA knockdown, in vivo neovascularization assays in immunocompromised mice\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with in vivo readout and receptor activation assays, single lab\",\n      \"pmids\": [\"25420155\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EphA3 depletion in gastric cancer cells inhibited HUVEC tube formation and migration as well as tumor angiogenesis in vivo, acting through the STAT3/VEGF signaling pathway.\",\n      \"method\": \"Lentivirus-mediated EphA3 knockdown, tube formation assay, migration assay, xenograft models with microvessel density analysis, STAT3/VEGF pathway western blotting\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — loss-of-function with defined pathway readout and in vivo confirmation, single lab\",\n      \"pmids\": [\"30066881\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"EPHA3 overexpression in SCLC cells reduced chemoresistance by increasing apoptosis and G0/G1 arrest with concomitant reduced phosphorylation of the PI3K/BMX/STAT3 signaling pathway; knockdown of EPHA3 promoted resistant phenotype with increased PI3K/BMX/STAT3 phosphorylation, and PI3K or BMX inhibitors reversed this resistance.\",\n      \"method\": \"EPHA3 overexpression and knockdown in SCLC cells, flow cytometry for apoptosis and cell cycle, pharmacological inhibitors (LY294002, LFM-A13), western blotting of PI3K/BMX/STAT3 pathway, in vivo tumor growth assays\",\n      \"journal\": \"Tumour biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — gain and loss-of-function with defined pathway readout and inhibitor rescue, single lab\",\n      \"pmids\": [\"27101199\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"EphA3 is expressed on CCR10+ mesenchymal progenitor cells in IPF lungs; antibody-mediated (ifabotuzumab) killing of EphA3+ cells significantly reduced CCR10+ cell numbers and ameliorated pulmonary fibrosis in humanized NSG mice.\",\n      \"method\": \"Human IPF cell characterization, intravenous injection of IPF cells into NSG mice, EphA3-targeted antibody (ifabotuzumab) treatment, histological fibrosis assessment\",\n      \"journal\": \"JCI insight\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo humanized mouse model with defined cellular and fibrosis readout, single study\",\n      \"pmids\": [\"33945505\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EphA3 knockdown via siRNA screening triggered cellular senescence; selected intracellular EPHA3 tumor-associated point mutations decrease receptor expression level and/or RTK activity, consistent with a tumor suppressor role.\",\n      \"method\": \"High-content siRNA screen for senescence markers (β-galactosidase, DNA damage, p53/p16 expression), RTK activity assays on EphA3 point mutants\",\n      \"journal\": \"Cell cycle\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — siRNA screen with defined senescence readout plus kinase activity assays on mutants, single lab\",\n      \"pmids\": [\"23324396\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EphA3 promotes axon elongation of nasal retinal ganglion cells by competing with EphA4 for ephrin-A binding, thereby reducing ephrin-A-mediated EphA4 forward signaling (decreased EphA4 and ephexin1 phosphorylation) and decreasing axonal filopodia density.\",\n      \"method\": \"Clustered EphA3-Fc treatment of retinal explants, in vitro and in vivo electroporation with dominant-negative EphA4, immunoprecipitation and western blot for EphA4/ephexin1 phosphorylation, PI-PLC shedding of ephrin-As\",\n      \"journal\": \"Experimental eye research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal approaches (gain and loss-of-function, ligand shedding) with defined phosphorylation and morphological readouts, single lab\",\n      \"pmids\": [\"30237102\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"EPHA3 contributes to epigenetic suppression of PTEN in radioresistant head and neck cancer cells via EPHA3/c-Myc signaling that upregulates DNMT1-mediated DNA methylation and EZH2-mediated H3K27me3 histone methylation at the PTEN locus.\",\n      \"method\": \"Western blotting for DNMT1, EZH2, H3K27me3, and PTEN; EPHA3 manipulation in radioresistant cell lines; correlation of signaling components\",\n      \"journal\": \"Biomolecules\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — correlative western blot data without direct ChIP or rescue experiments linking EPHA3 to specific epigenetic writers, single lab\",\n      \"pmids\": [\"33919657\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Androgen receptor (AR) induces EPHA3 expression by forming a transcriptional complex with SP1 that binds SP1 sites (−295 to −261) in the EPHA3 core promoter, as demonstrated by co-immunoprecipitation and chromatin immunoprecipitation assays.\",\n      \"method\": \"Co-immunoprecipitation, chromatin immunoprecipitation (ChIP), luciferase reporter assays with EPHA3 promoter truncations and SP1 site mutations, siRNA knockdown of AR and SP1\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP plus ChIP plus reporter assays with deletion and mutation analysis, single lab\",\n      \"pmids\": [\"29917167\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"EPHA3 overexpression in macrophages from endometriosis mice inhibits activation of the mTOR signaling pathway, downregulates bcl-2, upregulates autophagy markers (Atg3, LC3-II/LC3-I, Beclin1) and apoptosis markers (bax, fas), thereby promoting macrophage autophagy and apoptosis.\",\n      \"method\": \"Mouse endometriosis model, macrophage transfection with EPHA3 vector or siRNA, western blotting for mTOR pathway components and autophagy/apoptosis markers, autophagic lysosome visualization, flow cytometry for apoptosis\",\n      \"journal\": \"Bioscience reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — gain and loss-of-function with signaling readouts, but no direct mechanistic link between EPHA3 kinase activity and mTOR established, single lab\",\n      \"pmids\": [\"31262977\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EphA3 is a receptor tyrosine kinase whose kinase activity is autoinhibited by its unphosphorylated juxtamembrane segment (structurally defined by crystallography) and activated upon ephrin-A ligand binding and receptor oligomerization; it can also be silenced in cis by ephrinA5 co-expression independently of the canonical ligand-binding domain, or ligand-independently cleaved by presenilin/γ-secretase to generate an intracellular domain that binds NMIIA to promote axon elongation via RhoA inhibition; in cancer contexts, EphA3 acts as a context-dependent tumor suppressor (suppressing AKT signaling and inducing senescence) or tumor promoter (maintaining GBM tumor-initiating cells via MAPK signaling), with somatic cancer mutations disrupting kinase activity, ligand binding, or receptor surface localization.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"EphA3 is a receptor tyrosine kinase that transduces ephrin-A ligand signals to control cell migration, axon guidance, and tissue morphogenesis during development and in disease. Its kinase domain is autoinhibited by an unphosphorylated juxtamembrane segment that blocks the active site; autophosphorylation of two juxtamembrane tyrosines relieves this inhibition and enables signal transduction through downstream pathways including MAPK, AKT, and PI3K/BMX/STAT3 [PMID:18547520, PMID:22829656, PMID:23410976]. In the nervous system, EphA3 modulates axon elongation both through classical ephrin-A-dependent forward signaling and through ligand-independent presenilin/γ-secretase cleavage that generates an intracellular domain binding non-muscle myosin IIA to inhibit RhoA and promote cytoskeletal rearrangement [PMID:31577226, PMID:30237102]; cis-interaction with ephrinA5 silences EphA3 trans-signaling independently of its canonical ligand-binding domain [PMID:16491080]. In cancer, EphA3 functions as a context-dependent tumor suppressor—somatic mutations disrupt kinase activity, ligand binding, or surface localization, and wild-type re-expression suppresses AKT signaling and tumor growth—while in glioblastoma it maintains tumor-initiating cells via MAPK signaling [PMID:22242939, PMID:22829656, PMID:23410976].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Defining how EphA3 assembles signaling-competent oligomeric complexes with ephrin-A5 identified three essential receptor surface areas, one outside previously characterized interaction domains, establishing the structural requirements for ligand recognition and receptor clustering.\",\n      \"evidence\": \"Random mutagenesis screen of EphA3 in yeast with functional characterization of ephrin-A5 binding and signaling\",\n      \"pmids\": [\"14660665\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No full-length receptor-ligand co-crystal structure\", \"Contribution of the third novel contact area to in vivo signaling unclear\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Two parallel discoveries revealed that EphA3 can be silenced in cis by co-expressed ephrinA5 through a mechanism independent of the canonical ligand-binding domain, and that EphA3 is essential for cardiac mesenchymal transformation during septum and cushion development, establishing dual roles in axon guidance regulation and organogenesis.\",\n      \"evidence\": \"FRET, point mutagenesis, and retinal axon functional assays for cis-silencing; EphA3 knockout mouse analysis with endocardial cushion explant migration assays for cardiac role\",\n      \"pmids\": [\"16491080\", \"17046737\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the cis interaction is undefined\", \"Which downstream effectors mediate EphA3-driven EMT in the heart is unknown\", \"Whether cis-silencing operates in non-retinal tissues is untested\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Crystal structures of the EphA3 kinase domain resolved the autoinhibition mechanism: the unphosphorylated juxtamembrane segment directly occludes the active site, and autophosphorylation of two juxtamembrane tyrosines releases catalytic activity through a coupled allosteric pathway connecting the juxtamembrane region, activation loop, and catalytic residues.\",\n      \"evidence\": \"X-ray crystallography of wild-type and mutant EphA3 kinase domains with and without the juxtamembrane segment, combined with kinase activity assays\",\n      \"pmids\": [\"18547520\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structure of full-length EphA3 in an oligomeric signaling complex remains unknown\", \"Role of SAM domain in autoinhibition or activation not addressed\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Characterization of cancer-associated EphA3 mutations demonstrated that somatic mutations impair kinase activity, ligand binding, or surface expression, and that some act as dominant negatives; wild-type EphA3 re-expression suppressed AKT signaling and tumor growth, establishing EphA3 as an ephrin- and kinase-dependent tumor suppressor disrupted in multiple cancer types.\",\n      \"evidence\": \"Kinase activity, ephrin-binding, and surface localization assays on mutants; dominant-negative analysis; xenograft tumor growth and apoptosis assays with AKT pathway readout\",\n      \"pmids\": [\"22242939\", \"22829656\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How dominant-negative mutants suppress wild-type receptor at the structural level is unclear\", \"Patient-level genotype-phenotype correlations are limited\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"EphA3 was shown to have a context-dependent oncogenic role in glioblastoma, where it marks tumor-initiating cells, maintains stemness via MAPK signaling, and its depletion reduces tumorigenic potential—contrasting with its tumor suppressor function in other cancers and complemented by the finding that EphA3 knockdown induces senescence.\",\n      \"evidence\": \"EphA3 knockdown and EphA3+ cell depletion in GBM with MAPK signaling analysis; siRNA senescence screen with β-galactosidase, DNA damage, and p53/p16 readouts plus kinase assays on tumor-associated mutants\",\n      \"pmids\": [\"23410976\", \"23324396\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether EphA3 kinase activity or scaffolding function drives GBM stemness is unresolved\", \"Direct substrates mediating MAPK-dependent stemness maintenance are unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"A series of studies clarified EphA3's axon guidance mechanism: neurocan blocks NCAM/EphA3 clustering and kinase activation to modulate ephrin-A5-induced growth cone collapse, and EphA3 promotes nasal retinal axon elongation by competing with EphA4 for ephrin-A binding, reducing EphA4/ephexin1 phosphorylation—showing EphA3 acts as both a signal transducer and a competitive inhibitor of co-expressed Eph receptors.\",\n      \"evidence\": \"NCAM co-immunoprecipitation, EphA3 kinase activation, growth cone collapse assays in cortical interneurons; clustered EphA3-Fc treatment and dominant-negative EphA4 in retinal explants with phosphorylation readouts\",\n      \"pmids\": [\"29670169\", \"30237102\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether EphA3/EphA4 competition operates in non-retinal axon guidance contexts is untested\", \"Stoichiometric requirements for NCAM-EphA3 complex formation are undefined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Discovery of ligand-independent presenilin/γ-secretase cleavage of EphA3 revealed a non-canonical signaling mode: the released intracellular domain binds NMIIA, increases Ser1943 phosphorylation to promote filament disassembly, and drives axon elongation via RhoA inhibition, establishing a proteolytic activation pathway distinct from classical ephrin-dependent signaling.\",\n      \"evidence\": \"PS1/EphA3 KO neurons, proteomic identification of EphA3 ICD interactors, Ser1943 phosphorylation assays, ICD rescue experiments, pharmacological NMII inhibition\",\n      \"pmids\": [\"31577226\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The kinase or phosphatase directly responsible for NMIIA Ser1943 phosphorylation downstream of EphA3 ICD is unknown\", \"Whether γ-secretase cleavage of EphA3 occurs in non-neuronal tissues is untested\", \"Regulation of the initial extracellular domain shedding step is unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"EphA3 was identified as a therapeutic surface target on mesenchymal progenitor cells in idiopathic pulmonary fibrosis, where antibody-mediated killing of EphA3+ cells reduced fibrosis in a humanized mouse model, extending EphA3's disease relevance beyond cancer.\",\n      \"evidence\": \"Human IPF cell characterization, ifabotuzumab treatment in humanized NSG mice with fibrosis histological assessment\",\n      \"pmids\": [\"33945505\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether EphA3 kinase signaling drives the fibrotic phenotype or merely marks the pathogenic population is unclear\", \"Confirmation in independent cohorts and animal models needed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the full-length structure of EphA3 in signaling oligomers, the identity of direct EphA3 kinase substrates mediating MAPK- and AKT-dependent cellular outcomes, whether the γ-secretase cleavage pathway operates beyond neurons, and the structural basis by which cis ephrinA5 silences EphA3 signaling.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No full-length EphA3 signaling complex structure\", \"Direct kinase substrates largely unidentified\", \"In vivo relevance of γ-secretase cleavage in non-neuronal tissues untested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 5, 6, 13]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 14]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [2, 5, 8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 4, 6, 7, 11]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [3, 14]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [4, 5, 6, 13]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"EFNA5\",\n      \"EFNA1\",\n      \"NCAM1\",\n      \"MYH9\",\n      \"PSEN1\",\n      \"EPHA4\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}