{"gene":"MAGEA10","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":1999,"finding":"MAGE-A10 (MAGE-10) encodes a nuclear protein of approximately 72 kDa, as determined by in vitro translation, transient transfection experiments, immunocytochemistry of cultured melanoma cells, and Western blot analysis with a polyclonal antibody raised against a MAGE-10 peptide.","method":"In vitro translation, transient transfection, Western blot, immunocytochemistry","journal":"International journal of cancer","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (in vitro translation, transfection, Western blot, immunocytochemistry) in a single study establishing molecular weight and nuclear localization","pmids":["10446460"],"is_preprint":false},{"year":1999,"finding":"CTL clone from melanoma patient LB1751 recognizes the nonapeptide GLYDGMEHL (codons 254-262) derived from MAGE-A10, presented by HLA-A2.1, as demonstrated by TNF production upon stimulation with target cells expressing MAGE-A10 and lysis of tumor cell lines expressing HLA-A2.1 and MAGE-A10.","method":"CTL stimulation assay (TNF production), cytotoxicity assay with tumor cell lines expressing MAGE-A10 and HLA-A2.1","journal":"Journal of immunology (Baltimore, Md. : 1950)","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal functional assays (TNF production + cytotoxicity) identifying specific peptide epitope and HLA restriction element","pmids":["10352307"],"is_preprint":false},{"year":2004,"finding":"Novel alternative splice variants of MAGE-A10 mRNA were identified in primary esophageal adenocarcinoma tumors expressing MAGE-A10 protein, as revealed by RT-PCR and DNA sequencing.","method":"RT-PCR and DNA sequencing of primary tumor samples","journal":"Clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — RT-PCR and sequencing identifying novel splice variants, single lab but two methods","pmids":["15355897"],"is_preprint":false},{"year":2011,"finding":"Using a specific monoclonal antibody recognizing an epitope at the COOH terminus of MAGE-A10, the protein was localized as an intranuclear protein of apparent molecular weight 70 kDa, expressed in normal spermatogonia and spermatocytes, but in no other healthy tissue.","method":"Recombinant protein production, monoclonal antibody generation, immunohistochemistry on tissue microarray (>2,500 specimens)","journal":"International journal of cancer","confidence":"High","confidence_rationale":"Tier 2 / Strong — specific mAb generated and validated, immunohistochemistry on large tissue microarray confirming intranuclear localization across multiple tissue types","pmids":["21710496"],"is_preprint":false},{"year":2017,"finding":"shRNA-mediated knockdown of MAGE-A10 in oral squamous cell carcinoma cells (LN1 and LN2) impaired cell growth, increased cell-cell and cell-matrix adhesion, inhibited migration and invasion, and altered 3D microspheroid assembly. Concomitantly, knockdown activated E-cadherin expression and repressed N-cadherin and vimentin transcription, indicating MAGE-A10 promotes epithelial-mesenchymal transition (EMT) by regulating adhesion molecule expression.","method":"Lentiviral shRNA knockdown, wound healing assay, phagokinetic track motility assay, in vitro invasion assay (myogel matrix), 3D culture, RT-PCR for EMT markers","journal":"Biochemistry and biophysics reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with multiple orthogonal phenotypic readouts (migration, invasion, adhesion, EMT markers), single lab","pmids":["28955754"],"is_preprint":false},{"year":2022,"finding":"Crystal structures of parental and affinity-enhanced TCRs targeting the MAGE-A10 peptide GLYDGMEHL in complex with HLA-A*02:01 revealed: (1) the HLA-bound MAGE-A10 peptide contains an intrachain non-covalent 'staple' between peptide Tyr3 and Glu7; (2) a Glu31-Asp mutation in βCDR1 generates a high-affinity TCR derivative by causing a rigid-body shift of the TRBV domain toward the pHLA; (3) differential cross-reactivity between TCR variants is explained by alterations in surface electrostatics and the size/geometry of TCR-peptide interfacial cavities.","method":"X-ray crystallography (four high-resolution crystal structures of TCR alone and TCR/pHLA complexes), site-directed mutagenesis","journal":"Journal for immunotherapy of cancer","confidence":"High","confidence_rationale":"Tier 1 / Strong — multiple high-resolution crystal structures with functional mutagenesis, directly establishing peptide conformation and molecular basis for affinity and specificity","pmids":["35851311"],"is_preprint":false},{"year":2023,"finding":"The intrinsically disordered N-terminal domain of MAGEA10, and specifically the first seven amino acids containing the linear motif PRAPKR, are required for MAGEA10 protein expression levels, aberrant migration in SDS-PAGE, and nuclear localization, as shown by deletion and point-mutation analyses. Aberrant gel migration and nuclear localization are independent properties. Masking the N-terminus with an epitope tag strongly reduced mobility in gel and expression in cells.","method":"Deletion mutagenesis, point-mutation analysis, SDS-PAGE, cellular transfection/expression assays, fluorescence microscopy for nuclear localization","journal":"Biomolecules","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple deletion and point mutants with orthogonal readouts (expression level, gel migration, nuclear localization) in a single study","pmids":["38136576"],"is_preprint":false},{"year":2020,"finding":"Treatment of lung cancer cell lines with 5-aza-2'-deoxycytidine (DAC), a DNA methyltransferase inhibitor, induced MAGE-A10 expression, demonstrating that MAGE-A10 expression is regulated by DNA methylation (epigenetic silencing). MAGE-A10-specific CTLs showed enhanced cytotoxicity against DAC-treated cells expressing MAGE-A10.","method":"DAC treatment of lung cancer cell lines, real-time PCR and Western blot for MAGE-A10 expression, CCK-8 cytotoxicity assay","journal":"Translational cancer research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — pharmacological demethylation with two orthogonal expression readouts (PCR + Western blot), single lab","pmids":["35117468"],"is_preprint":false},{"year":2025,"finding":"Knockdown of MAGE-A10 in gastric cancer cells significantly reduced proliferation, migration, and invasion, and GSEA indicated that high MAGE-A10 expression is closely associated with histone deacetylase binding. Knockdown also affected expression of cell adhesion molecule cadherins.","method":"siRNA/shRNA knockdown in gastric cancer cell lines, proliferation/migration/invasion assays, GSEA, cadherin expression analysis","journal":"Scientific reports","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, GSEA is computational, cadherin regulation inferred without direct mechanistic follow-up","pmids":["40890425"],"is_preprint":false}],"current_model":"MAGEA10 encodes an intrinsically disordered N-terminus-containing nuclear protein (~70-72 kDa) whose nuclear localization and expression are controlled by the N-terminal PRAPKR motif; its expression is epigenetically silenced by DNA methylation; it presents the HLA-A*02:01-restricted nonapeptide GLYDGMEHL to cytotoxic T lymphocytes; and it promotes epithelial-mesenchymal transition by upregulating N-cadherin and vimentin while suppressing E-cadherin, thereby supporting cancer cell migration and invasion."},"narrative":{"mechanistic_narrative":"MAGEA10 is a cancer-testis antigen encoding an intranuclear protein of approximately 70–72 kDa whose normal expression is restricted to spermatogonia and spermatocytes, with aberrant re-expression in diverse tumors [PMID:10446460, PMID:21710496]. Its silencing in normal somatic tissue is enforced epigenetically: treatment of cancer cells with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine induces MAGEA10 expression [PMID:35117468]. The protein contains an intrinsically disordered N-terminus whose first seven residues, harboring a PRAPKR linear motif, are required for protein expression level and nuclear localization, two properties that are genetically separable [PMID:38136576]. Functionally, MAGEA10 promotes epithelial-mesenchymal transition: loss-of-function in oral squamous cell carcinoma cells activates E-cadherin while repressing N-cadherin and vimentin, with concomitant impairment of migration, invasion, and adhesion [PMID:28955754]. As an immunotherapy target, MAGEA10 yields the HLA-A*02:01-restricted nonapeptide GLYDGMEHL recognized by cytotoxic T lymphocytes, and crystal structures of TCR/peptide-HLA complexes have defined the bound peptide conformation and the molecular basis of engineered TCR affinity and specificity [PMID:10352307, PMID:35851311]. Beyond these roles, no enzymatic activity, direct binding partner, or detailed mechanism by which MAGEA10 regulates adhesion-molecule transcription has been characterized in the available corpus.","teleology":[{"year":1999,"claim":"Established the basic identity of the MAGEA10 product, defining it as a nuclear protein of defined size, a prerequisite for any functional study.","evidence":"In vitro translation, transfection, Western blot, and immunocytochemistry in melanoma cells","pmids":["10446460"],"confidence":"High","gaps":["No functional activity assigned to the protein","No domain structure or interaction partners identified"]},{"year":1999,"claim":"Answered whether MAGEA10 is an immunologically presented tumor antigen by identifying a specific HLA-A2.1-restricted epitope recognized by patient-derived CTLs, opening an immunotherapy rationale.","evidence":"CTL TNF-production and cytotoxicity assays against tumor lines expressing MAGEA10 and HLA-A2.1","pmids":["10352307"],"confidence":"High","gaps":["Did not determine the protein's cellular function","Epitope processing details not resolved"]},{"year":2004,"claim":"Showed MAGEA10 transcripts undergo alternative splicing in tumors, raising the question of isoform diversity in antigen expression.","evidence":"RT-PCR and DNA sequencing of primary esophageal adenocarcinoma samples","pmids":["15355897"],"confidence":"Medium","gaps":["Functional consequence of splice variants unknown","Protein-level confirmation of variants not established"]},{"year":2011,"claim":"Defined the normal tissue restriction of MAGEA10, confirming intranuclear localization and confining expression to male germ cells, the hallmark of a cancer-testis antigen.","evidence":"Validated monoclonal antibody and immunohistochemistry across a >2,500-specimen tissue microarray","pmids":["21710496"],"confidence":"High","gaps":["No mechanism for nuclear retention identified","Function in spermatogenesis not addressed"]},{"year":2017,"claim":"Provided the first functional role for MAGEA10, showing it drives epithelial-mesenchymal transition and tumor cell motility through regulation of cadherin and vimentin expression.","evidence":"Lentiviral shRNA knockdown in oral squamous cell carcinoma cells with migration, invasion, adhesion, 3D culture, and EMT-marker readouts","pmids":["28955754"],"confidence":"Medium","gaps":["Mechanism linking MAGEA10 to adhesion-gene transcription not defined","Single lab and single cancer type","No direct molecular partners identified"]},{"year":2020,"claim":"Established that MAGEA10's tumor-restricted expression is governed by DNA methylation, explaining its silencing in somatic tissue and offering a route to pharmacologically enhance antigen presentation.","evidence":"5-aza-2'-deoxycytidine treatment of lung cancer lines with PCR/Western readouts and CTL cytotoxicity assay","pmids":["35117468"],"confidence":"Medium","gaps":["Specific methylated regulatory elements not mapped","Single lab pharmacological inference"]},{"year":2022,"claim":"Resolved the structural basis of MAGEA10 antigen recognition, defining the HLA-bound peptide conformation and how TCR engineering modulates affinity and cross-reactivity for therapeutic targeting.","evidence":"Four high-resolution X-ray crystal structures of TCR and TCR/pHLA complexes with site-directed mutagenesis","pmids":["35851311"],"confidence":"High","gaps":["Addresses the presented peptide, not full-length protein function","In vivo efficacy of engineered TCRs not established here"]},{"year":2023,"claim":"Mapped the determinants of MAGEA10 expression and nuclear localization to an intrinsically disordered N-terminal PRAPKR motif, showing these properties are genetically separable and N-terminus-dependent.","evidence":"Deletion and point mutagenesis with SDS-PAGE, expression, and fluorescence-microscopy localization readouts","pmids":["38136576"],"confidence":"High","gaps":["Trans-acting factors recognizing the PRAPKR motif unidentified","Mechanism coupling the motif to protein stability unresolved"]},{"year":2025,"claim":"Extended MAGEA10's pro-tumor role to gastric cancer and linked it to histone deacetylase binding and cadherin regulation, hinting at a chromatin-associated mechanism.","evidence":"siRNA/shRNA knockdown in gastric cancer lines with proliferation/migration/invasion assays, GSEA, and cadherin analysis","pmids":["40890425"],"confidence":"Low","gaps":["HDAC association is computational (GSEA), not biochemically validated","Direct interaction with HDACs not demonstrated","Cadherin regulation inferred without mechanistic follow-up"]},{"year":null,"claim":"The biochemical activity of nuclear MAGEA10 and the direct molecular mechanism by which it represses E-cadherin and activates mesenchymal genes remain undefined.","evidence":"No reconstituted activity assay or direct partner identification in the available corpus","pmids":[],"confidence":"Low","gaps":["No enzymatic or binding activity assigned","No validated direct protein partner","Link between nuclear localization and transcriptional output unestablished"]}],"mechanism_profile":{"molecular_activity":[],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,3,6]}],"pathway":[],"complexes":[],"partners":[],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P43363","full_name":"Melanoma-associated antigen 10","aliases":["Cancer/testis antigen 1.10","CT1.10","MAGE-10 antigen"],"length_aa":369,"mass_kda":40.8,"function":"Not known, though may play a role in embryonal development and tumor transformation or aspects of tumor progression","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/P43363/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MAGEA10","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MAGEA10","total_profiled":1310},"omim":[{"mim_id":"300470","title":"MELANOMA ANTIGEN, FAMILY D, 2; MAGED2","url":"https://www.omim.org/entry/300470"},{"mim_id":"300343","title":"MELANOMA ANTIGEN, FAMILY A, 10; MAGEA10","url":"https://www.omim.org/entry/300343"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"placenta","ntpm":8.9},{"tissue":"testis","ntpm":2.4}],"url":"https://www.proteinatlas.org/search/MAGEA10"},"hgnc":{"alias_symbol":["MGC10599","CT1.10"],"prev_symbol":["MAGE10"]},"alphafold":{"accession":"P43363","domains":[{"cath_id":"1.10.10.1200","chopping":"132-209","consensus_level":"high","plddt":93.6886,"start":132,"end":209},{"cath_id":"1.10.10.1210","chopping":"224-307","consensus_level":"high","plddt":94.2993,"start":224,"end":307}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P43363","model_url":"https://alphafold.ebi.ac.uk/files/AF-P43363-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P43363-F1-predicted_aligned_error_v6.png","plddt_mean":72.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MAGEA10","jax_strain_url":"https://www.jax.org/strain/search?query=MAGEA10"},"sequence":{"accession":"P43363","fasta_url":"https://rest.uniprot.org/uniprotkb/P43363.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P43363/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P43363"}},"corpus_meta":[{"pmid":"15728523","id":"PMC_15728523","title":"MAGE-A1-, MAGE-A10-, and gp100-derived peptides are immunogenic when combined with granulocyte-macrophage colony-stimulating factor and montanide ISA-51 adjuvant and administered as part of a multipeptide vaccine for melanoma.","date":"2005","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/15728523","citation_count":93,"is_preprint":false},{"pmid":"15661935","id":"PMC_15661935","title":"Naturally acquired MAGE-A10- and SSX-2-specific CD8+ T cell responses in patients with hepatocellular carcinoma.","date":"2005","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/15661935","citation_count":75,"is_preprint":false},{"pmid":"30713784","id":"PMC_30713784","title":"Affinity-enhanced T-cell receptors for adoptive T-cell therapy targeting MAGE-A10: strategy for selection of an optimal candidate.","date":"2018","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/30713784","citation_count":56,"is_preprint":false},{"pmid":"10352307","id":"PMC_10352307","title":"Cytolytic T lymphocytes recognize an antigen encoded by MAGE-A10 on a human melanoma.","date":"1999","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/10352307","citation_count":55,"is_preprint":false},{"pmid":"21710496","id":"PMC_21710496","title":"MAGE-A10 is a nuclear protein frequently expressed in high percentages of tumor cells in lung, skin and urothelial malignancies.","date":"2011","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/21710496","citation_count":51,"is_preprint":false},{"pmid":"15355897","id":"PMC_15355897","title":"Melanoma-associated antigens in esophageal adenocarcinoma: identification of novel MAGE-A10 splice variants.","date":"2004","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/15355897","citation_count":48,"is_preprint":false},{"pmid":"35086946","id":"PMC_35086946","title":"Phase I clinical trial evaluating the safety and efficacy of ADP-A2M10 SPEAR T cells in patients with MAGE-A10+ advanced non-small cell lung cancer.","date":"2022","source":"Journal for immunotherapy of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/35086946","citation_count":44,"is_preprint":false},{"pmid":"10446460","id":"PMC_10446460","title":"cDNA and protein characterization of human MAGE-10.","date":"1999","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/10446460","citation_count":38,"is_preprint":false},{"pmid":"23125074","id":"PMC_23125074","title":"MAGE-A10 cancer/testis antigen is highly expressed in high-grade non-muscle-invasive bladder carcinomas.","date":"2012","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/23125074","citation_count":28,"is_preprint":false},{"pmid":"22116775","id":"PMC_22116775","title":"High expression of MAGE-A10 cancer-testis antigen in triple-negative breast cancer.","date":"2011","source":"Medical oncology (Northwood, London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/22116775","citation_count":22,"is_preprint":false},{"pmid":"35372008","id":"PMC_35372008","title":"Phase 1 Clinical Trial Evaluating the Safety and Anti-Tumor Activity of ADP-A2M10 SPEAR T-Cells in Patients With MAGE-A10+ Head and Neck, Melanoma, or Urothelial Tumors.","date":"2022","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/35372008","citation_count":15,"is_preprint":false},{"pmid":"29928403","id":"PMC_29928403","title":"Antibody response against cancer-testis antigens MAGEA4 and MAGEA10 in patients with melanoma.","date":"2018","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/29928403","citation_count":10,"is_preprint":false},{"pmid":"28955754","id":"PMC_28955754","title":"Suppression of MAGE-A10 alters the metastatic phenotype of tongue squamous cell carcinoma cells.","date":"2017","source":"Biochemistry and biophysics reports","url":"https://pubmed.ncbi.nlm.nih.gov/28955754","citation_count":9,"is_preprint":false},{"pmid":"35851311","id":"PMC_35851311","title":"Structural insights into engineering a T-cell receptor targeting MAGE-A10 with higher affinity and specificity for cancer immunotherapy.","date":"2022","source":"Journal for immunotherapy of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/35851311","citation_count":7,"is_preprint":false},{"pmid":"32965606","id":"PMC_32965606","title":"MAGEA10 expression is a predictive marker of early hepatic recurrence after curative gastrectomy for gastric and gastroesophageal junction cancer.","date":"2020","source":"Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association","url":"https://pubmed.ncbi.nlm.nih.gov/32965606","citation_count":6,"is_preprint":false},{"pmid":"35117468","id":"PMC_35117468","title":"5-aza-2'-deoxycytidine (DAC) treatment induces the MAGE-A10 expression and improves the cytotoxicity of MAGE-A10-specific CTLs in lung cancer cells.","date":"2020","source":"Translational cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/35117468","citation_count":5,"is_preprint":false},{"pmid":"38136576","id":"PMC_38136576","title":"How the Intrinsically Disordered N-Terminus of Cancer/Testis Antigen MAGEA10 Is Responsible for Its Expression, Nuclear Localisation and Aberrant Migration.","date":"2023","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/38136576","citation_count":1,"is_preprint":false},{"pmid":"40379175","id":"PMC_40379175","title":"Identification of MAGE-A10 specific T cell receptor promising in immunotherapy of hepatocellular carcinoma.","date":"2025","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/40379175","citation_count":1,"is_preprint":false},{"pmid":"40890425","id":"PMC_40890425","title":"Comprehensive analysis of MAGE-A10 in pan-cancer and its validation in gastric cancer.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/40890425","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13055,"output_tokens":2358,"usd":0.037268,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9516,"output_tokens":2765,"usd":0.058353,"stage2_stop_reason":"end_turn"},"total_usd":0.095621,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"MAGE-A10 (MAGE-10) encodes a nuclear protein of approximately 72 kDa, as determined by in vitro translation, transient transfection experiments, immunocytochemistry of cultured melanoma cells, and Western blot analysis with a polyclonal antibody raised against a MAGE-10 peptide.\",\n      \"method\": \"In vitro translation, transient transfection, Western blot, immunocytochemistry\",\n      \"journal\": \"International journal of cancer\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (in vitro translation, transfection, Western blot, immunocytochemistry) in a single study establishing molecular weight and nuclear localization\",\n      \"pmids\": [\"10446460\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"CTL clone from melanoma patient LB1751 recognizes the nonapeptide GLYDGMEHL (codons 254-262) derived from MAGE-A10, presented by HLA-A2.1, as demonstrated by TNF production upon stimulation with target cells expressing MAGE-A10 and lysis of tumor cell lines expressing HLA-A2.1 and MAGE-A10.\",\n      \"method\": \"CTL stimulation assay (TNF production), cytotoxicity assay with tumor cell lines expressing MAGE-A10 and HLA-A2.1\",\n      \"journal\": \"Journal of immunology (Baltimore, Md. : 1950)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal functional assays (TNF production + cytotoxicity) identifying specific peptide epitope and HLA restriction element\",\n      \"pmids\": [\"10352307\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Novel alternative splice variants of MAGE-A10 mRNA were identified in primary esophageal adenocarcinoma tumors expressing MAGE-A10 protein, as revealed by RT-PCR and DNA sequencing.\",\n      \"method\": \"RT-PCR and DNA sequencing of primary tumor samples\",\n      \"journal\": \"Clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — RT-PCR and sequencing identifying novel splice variants, single lab but two methods\",\n      \"pmids\": [\"15355897\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Using a specific monoclonal antibody recognizing an epitope at the COOH terminus of MAGE-A10, the protein was localized as an intranuclear protein of apparent molecular weight 70 kDa, expressed in normal spermatogonia and spermatocytes, but in no other healthy tissue.\",\n      \"method\": \"Recombinant protein production, monoclonal antibody generation, immunohistochemistry on tissue microarray (>2,500 specimens)\",\n      \"journal\": \"International journal of cancer\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — specific mAb generated and validated, immunohistochemistry on large tissue microarray confirming intranuclear localization across multiple tissue types\",\n      \"pmids\": [\"21710496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"shRNA-mediated knockdown of MAGE-A10 in oral squamous cell carcinoma cells (LN1 and LN2) impaired cell growth, increased cell-cell and cell-matrix adhesion, inhibited migration and invasion, and altered 3D microspheroid assembly. Concomitantly, knockdown activated E-cadherin expression and repressed N-cadherin and vimentin transcription, indicating MAGE-A10 promotes epithelial-mesenchymal transition (EMT) by regulating adhesion molecule expression.\",\n      \"method\": \"Lentiviral shRNA knockdown, wound healing assay, phagokinetic track motility assay, in vitro invasion assay (myogel matrix), 3D culture, RT-PCR for EMT markers\",\n      \"journal\": \"Biochemistry and biophysics reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with multiple orthogonal phenotypic readouts (migration, invasion, adhesion, EMT markers), single lab\",\n      \"pmids\": [\"28955754\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Crystal structures of parental and affinity-enhanced TCRs targeting the MAGE-A10 peptide GLYDGMEHL in complex with HLA-A*02:01 revealed: (1) the HLA-bound MAGE-A10 peptide contains an intrachain non-covalent 'staple' between peptide Tyr3 and Glu7; (2) a Glu31-Asp mutation in βCDR1 generates a high-affinity TCR derivative by causing a rigid-body shift of the TRBV domain toward the pHLA; (3) differential cross-reactivity between TCR variants is explained by alterations in surface electrostatics and the size/geometry of TCR-peptide interfacial cavities.\",\n      \"method\": \"X-ray crystallography (four high-resolution crystal structures of TCR alone and TCR/pHLA complexes), site-directed mutagenesis\",\n      \"journal\": \"Journal for immunotherapy of cancer\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — multiple high-resolution crystal structures with functional mutagenesis, directly establishing peptide conformation and molecular basis for affinity and specificity\",\n      \"pmids\": [\"35851311\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The intrinsically disordered N-terminal domain of MAGEA10, and specifically the first seven amino acids containing the linear motif PRAPKR, are required for MAGEA10 protein expression levels, aberrant migration in SDS-PAGE, and nuclear localization, as shown by deletion and point-mutation analyses. Aberrant gel migration and nuclear localization are independent properties. Masking the N-terminus with an epitope tag strongly reduced mobility in gel and expression in cells.\",\n      \"method\": \"Deletion mutagenesis, point-mutation analysis, SDS-PAGE, cellular transfection/expression assays, fluorescence microscopy for nuclear localization\",\n      \"journal\": \"Biomolecules\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple deletion and point mutants with orthogonal readouts (expression level, gel migration, nuclear localization) in a single study\",\n      \"pmids\": [\"38136576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Treatment of lung cancer cell lines with 5-aza-2'-deoxycytidine (DAC), a DNA methyltransferase inhibitor, induced MAGE-A10 expression, demonstrating that MAGE-A10 expression is regulated by DNA methylation (epigenetic silencing). MAGE-A10-specific CTLs showed enhanced cytotoxicity against DAC-treated cells expressing MAGE-A10.\",\n      \"method\": \"DAC treatment of lung cancer cell lines, real-time PCR and Western blot for MAGE-A10 expression, CCK-8 cytotoxicity assay\",\n      \"journal\": \"Translational cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — pharmacological demethylation with two orthogonal expression readouts (PCR + Western blot), single lab\",\n      \"pmids\": [\"35117468\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Knockdown of MAGE-A10 in gastric cancer cells significantly reduced proliferation, migration, and invasion, and GSEA indicated that high MAGE-A10 expression is closely associated with histone deacetylase binding. Knockdown also affected expression of cell adhesion molecule cadherins.\",\n      \"method\": \"siRNA/shRNA knockdown in gastric cancer cell lines, proliferation/migration/invasion assays, GSEA, cadherin expression analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, GSEA is computational, cadherin regulation inferred without direct mechanistic follow-up\",\n      \"pmids\": [\"40890425\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MAGEA10 encodes an intrinsically disordered N-terminus-containing nuclear protein (~70-72 kDa) whose nuclear localization and expression are controlled by the N-terminal PRAPKR motif; its expression is epigenetically silenced by DNA methylation; it presents the HLA-A*02:01-restricted nonapeptide GLYDGMEHL to cytotoxic T lymphocytes; and it promotes epithelial-mesenchymal transition by upregulating N-cadherin and vimentin while suppressing E-cadherin, thereby supporting cancer cell migration and invasion.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MAGEA10 is a cancer-testis antigen encoding an intranuclear protein of approximately 70–72 kDa whose normal expression is restricted to spermatogonia and spermatocytes, with aberrant re-expression in diverse tumors [#0, #3]. Its silencing in normal somatic tissue is enforced epigenetically: treatment of cancer cells with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine induces MAGEA10 expression [#7]. The protein contains an intrinsically disordered N-terminus whose first seven residues, harboring a PRAPKR linear motif, are required for protein expression level and nuclear localization, two properties that are genetically separable [#6]. Functionally, MAGEA10 promotes epithelial-mesenchymal transition: loss-of-function in oral squamous cell carcinoma cells activates E-cadherin while repressing N-cadherin and vimentin, with concomitant impairment of migration, invasion, and adhesion [#4]. As an immunotherapy target, MAGEA10 yields the HLA-A*02:01-restricted nonapeptide GLYDGMEHL recognized by cytotoxic T lymphocytes, and crystal structures of TCR/peptide-HLA complexes have defined the bound peptide conformation and the molecular basis of engineered TCR affinity and specificity [#1, #5]. Beyond these roles, no enzymatic activity, direct binding partner, or detailed mechanism by which MAGEA10 regulates adhesion-molecule transcription has been characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Established the basic identity of the MAGEA10 product, defining it as a nuclear protein of defined size, a prerequisite for any functional study.\",\n      \"evidence\": \"In vitro translation, transfection, Western blot, and immunocytochemistry in melanoma cells\",\n      \"pmids\": [\"10446460\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No functional activity assigned to the protein\", \"No domain structure or interaction partners identified\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Answered whether MAGEA10 is an immunologically presented tumor antigen by identifying a specific HLA-A2.1-restricted epitope recognized by patient-derived CTLs, opening an immunotherapy rationale.\",\n      \"evidence\": \"CTL TNF-production and cytotoxicity assays against tumor lines expressing MAGEA10 and HLA-A2.1\",\n      \"pmids\": [\"10352307\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not determine the protein's cellular function\", \"Epitope processing details not resolved\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Showed MAGEA10 transcripts undergo alternative splicing in tumors, raising the question of isoform diversity in antigen expression.\",\n      \"evidence\": \"RT-PCR and DNA sequencing of primary esophageal adenocarcinoma samples\",\n      \"pmids\": [\"15355897\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of splice variants unknown\", \"Protein-level confirmation of variants not established\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined the normal tissue restriction of MAGEA10, confirming intranuclear localization and confining expression to male germ cells, the hallmark of a cancer-testis antigen.\",\n      \"evidence\": \"Validated monoclonal antibody and immunohistochemistry across a >2,500-specimen tissue microarray\",\n      \"pmids\": [\"21710496\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No mechanism for nuclear retention identified\", \"Function in spermatogenesis not addressed\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Provided the first functional role for MAGEA10, showing it drives epithelial-mesenchymal transition and tumor cell motility through regulation of cadherin and vimentin expression.\",\n      \"evidence\": \"Lentiviral shRNA knockdown in oral squamous cell carcinoma cells with migration, invasion, adhesion, 3D culture, and EMT-marker readouts\",\n      \"pmids\": [\"28955754\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking MAGEA10 to adhesion-gene transcription not defined\", \"Single lab and single cancer type\", \"No direct molecular partners identified\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established that MAGEA10's tumor-restricted expression is governed by DNA methylation, explaining its silencing in somatic tissue and offering a route to pharmacologically enhance antigen presentation.\",\n      \"evidence\": \"5-aza-2'-deoxycytidine treatment of lung cancer lines with PCR/Western readouts and CTL cytotoxicity assay\",\n      \"pmids\": [\"35117468\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Specific methylated regulatory elements not mapped\", \"Single lab pharmacological inference\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Resolved the structural basis of MAGEA10 antigen recognition, defining the HLA-bound peptide conformation and how TCR engineering modulates affinity and cross-reactivity for therapeutic targeting.\",\n      \"evidence\": \"Four high-resolution X-ray crystal structures of TCR and TCR/pHLA complexes with site-directed mutagenesis\",\n      \"pmids\": [\"35851311\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Addresses the presented peptide, not full-length protein function\", \"In vivo efficacy of engineered TCRs not established here\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Mapped the determinants of MAGEA10 expression and nuclear localization to an intrinsically disordered N-terminal PRAPKR motif, showing these properties are genetically separable and N-terminus-dependent.\",\n      \"evidence\": \"Deletion and point mutagenesis with SDS-PAGE, expression, and fluorescence-microscopy localization readouts\",\n      \"pmids\": [\"38136576\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Trans-acting factors recognizing the PRAPKR motif unidentified\", \"Mechanism coupling the motif to protein stability unresolved\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Extended MAGEA10's pro-tumor role to gastric cancer and linked it to histone deacetylase binding and cadherin regulation, hinting at a chromatin-associated mechanism.\",\n      \"evidence\": \"siRNA/shRNA knockdown in gastric cancer lines with proliferation/migration/invasion assays, GSEA, and cadherin analysis\",\n      \"pmids\": [\"40890425\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"HDAC association is computational (GSEA), not biochemically validated\", \"Direct interaction with HDACs not demonstrated\", \"Cadherin regulation inferred without mechanistic follow-up\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The biochemical activity of nuclear MAGEA10 and the direct molecular mechanism by which it represses E-cadherin and activates mesenchymal genes remain undefined.\",\n      \"evidence\": \"No reconstituted activity assay or direct partner identification in the available corpus\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No enzymatic or binding activity assigned\", \"No validated direct protein partner\", \"Link between nuclear localization and transcriptional output unestablished\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 3, 6]}\n    ],\n    \"pathway\": [],\n    \"complexes\": [],\n    \"partners\": [],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":5,"faith_pct":80.0}}