{"gene":"ZBTB10","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":2019,"finding":"ZBTB10 directly binds the telomeric variant repeat sequence TTGGGG via its two C2H2 zinc fingers with nanomolar affinity, co-localizes with a subset of telomeres in ALT-positive U2OS cells, and interacts with TRF2/RAP1 via the N-terminal region of TRF2.","method":"In vitro binding assays, co-localization imaging in ALT cells, co-immunoprecipitation with TRF2/RAP1","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — direct binding demonstrated in vitro with affinity measurement, domain mapping, reciprocal Co-IP, and cellular co-localization, multiple orthogonal methods in single study","pmids":["30629181"],"is_preprint":false},{"year":2023,"finding":"Crystal structure of human ZBTB10 ZF1-2 in complex with double-stranded TTGGGG DNA revealed the molecular basis of sequence-specific recognition; a single Arg767Gln substitution shifts preference from TTGGGG to the canonical TTAGGG repeat, as confirmed by co-crystal structure and calorimetric analysis.","method":"X-ray crystallography, isothermal titration calorimetry, site-directed mutagenesis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — atomic-resolution crystal structures of wild-type and mutant complexes plus calorimetric validation in one rigorous study","pmids":["36657642"],"is_preprint":false},{"year":2010,"finding":"ZBTB10 functions as a transcriptional repressor of Sp1, Sp3, and Sp4 in colon cancer cells; its expression is constitutively suppressed by miR-27a, and ROS-mediated downregulation of miR-27a leads to ZBTB10 induction and subsequent repression of Sp-regulated genes (cyclin D1, c-Met, EGFR, bcl-2, survivin, VEGF/VEGFRs).","method":"RNA interference knockdown, ZBTB10 expression plasmid transfection, RT-PCR, Western blot, miR-27a mimic/inhibitor experiments, antioxidant rescue (glutathione co-treatment)","journal":"Molecular cancer research : MCR","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — multiple orthogonal approaches (RNAi, overexpression, mimic/inhibitor, chemical rescue) in single lab across multiple cell lines","pmids":["21156786"],"is_preprint":false},{"year":2021,"finding":"Zbtb10 is required for NF-κB activation in cDC1 dendritic cells; Zbtb10 knockdown enhanced expression of NF-κB repressing factor (NKRF), abrogated p65 and RelB nuclear translocation, and suppressed expression of co-stimulatory molecules CD80/CD86 and cytokines IL-12, IL-6, and IL-10, thereby impairing CD4+ T cell Th1 differentiation.","method":"shRNA knockdown in Mutu-DC line, global transcriptome analysis, Western blot for p65/RelB nuclear translocation, T cell co-culture functional assays","journal":"European journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KD with defined cellular and molecular phenotype plus mechanistic pathway identification (NKRF/NF-κB), single lab","pmids":["33527393"],"is_preprint":false},{"year":2022,"finding":"ZBTB10 directly transcriptionally represses PKLR expression in prostate cancer cells; loss of ZBTB10 following androgen-deprivation therapy activates PKLR, enhancing glycolysis and neuroendocrine differentiation.","method":"Loss-of-function/gain-of-function experiments, RT-PCR, Western blot, transcriptional reporter assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — direct transcriptional regulation demonstrated with overexpression/knockdown and functional readouts in single lab","pmids":["35306527"],"is_preprint":false},{"year":2023,"finding":"ZBTB10 directly binds the HK1 promoter and activates HK1 transcription, as demonstrated by chromatin immunoprecipitation and luciferase reporter assay; ZBTB10 knockdown decreased HK1 expression and ZBTB10 overexpression increased it in laryngeal cancer cells and 293T cells.","method":"ChIP assay, luciferase reporter assay, shRNA knockdown, overexpression in cell lines","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct promoter binding and transcriptional activation confirmed by ChIP and reporter assay, single lab","pmids":["37834257"],"is_preprint":false},{"year":2025,"finding":"ZBTB10 directly binds a specific response element in the ARRDC3 promoter to enhance ARRDC3 expression; elevated ARRDC3 interacts with β4-integrin (ITGB4) and promotes its ubiquitination and degradation, leading to reduced PI3K/AKT phosphorylation and suppression of gastric cancer progression.","method":"ChIP assay, co-immunoprecipitation, luciferase reporter assay, overexpression/knockdown, xenograft assays, phospho-proteomic profiling","journal":"Cellular oncology (Dordrecht, Netherlands)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (ChIP, Co-IP, reporter assay, in vivo xenograft) in single lab establishing direct promoter binding and downstream protein interactions","pmids":["39873948"],"is_preprint":false},{"year":2025,"finding":"miR-582-5p (delivered via M1 macrophage exosomes) directly binds the 3′UTR of ZBTB10 and suppresses its expression; ZBTB10 suppression leads to increased H3K27ac modification of the TFR1 promoter, enhanced TFR1 transcription, and ferroptosis in renal tubular epithelial cells.","method":"Dual-luciferase reporter assay (3′UTR), miR-582-5p overexpression, RT-qPCR, Western blot, ChIP for H3K27ac","journal":"Archives of medical science : AMS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — 3′UTR luciferase validation plus H3K27ac ChIP linking ZBTB10 loss to TFR1 epigenetic regulation, single lab with multiple methods","pmids":["42110628"],"is_preprint":false},{"year":2012,"finding":"miR-27a constitutively inhibits ZBTB10 expression; in ErbB2-overexpressing breast cancer cells, betulinic acid induces ZBTB10 and represses Sp1/Sp3/Sp4 and their downstream target ErbB2 (via YY1) through cannabinoid receptor CB1 and CB2-dependent mechanisms.","method":"miR-27a mimic transfection, ZBTB10 overexpression, CB1/CB2 receptor antagonists, RNA interference of Sp1/Sp3/Sp4","journal":"Molecular cancer therapeutics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — receptor-dependency established pharmacologically with multiple cell lines and RNAi rescue, single lab","pmids":["22553354"],"is_preprint":false},{"year":2022,"finding":"miR-361-5p directly targets the 3′UTR of ZBTB10 (confirmed by dual luciferase assay) and suppresses its expression; ZBTB10 overexpression reverses the pro-proliferative, anti-apoptotic effects of miR-361-5p in rheumatoid arthritis fibroblast-like synoviocytes.","method":"Dual luciferase reporter assay, ZBTB10 overexpression rescue experiments, flow cytometry, Western blot","journal":"Autoimmunity","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — 3′UTR luciferase validation plus rescue experiment, single lab","pmids":["35608340"],"is_preprint":false}],"current_model":"ZBTB10 is a BTB-ZF transcription factor that uses its tandem C2H2 zinc fingers to bind the telomeric variant repeat TTGGGG with nanomolar affinity (crystal structure resolved) and interacts with TRF2/RAP1 at ALT telomeres; in the nucleus it directly represses Sp1/Sp3/Sp4 transcription factors (with downstream effects on VEGF, survivin, ErbB2, and MDR1), directly activates target gene promoters (HK1, ARRDC3), and transcriptionally represses PKLR to control glycolytic and neuroendocrine programs, while its own expression is post-transcriptionally suppressed by multiple miRNAs (miR-27a, miR-361-5p, miR-582-5p) and it supports NF-κB-dependent activation of dendritic cells."},"narrative":{"mechanistic_narrative":"ZBTB10 is a BTB-ZF transcription factor that uses tandem C2H2 zinc fingers to read specific DNA sequences and to regulate transcriptional programs across telomere biology, cancer metabolism, and immune cell activation [PMID:30629181, PMID:21156786]. At telomeres, ZBTB10 binds the variant repeat TTGGGG with nanomolar affinity, co-localizes with a subset of telomeres in ALT-positive cells, and associates with the shelterin proteins TRF2/RAP1; a crystal structure of its ZF1-2 domain bound to TTGGGG defines the basis of sequence-specific recognition, where a single Arg767Gln substitution redirects preference toward the canonical TTAGGG repeat [PMID:30629181, PMID:36657642]. As a transcriptional regulator it acts bidirectionally: it represses the Sp transcription factor family (Sp1/Sp3/Sp4) and their downstream targets including survivin, VEGF, and ErbB2 [PMID:21156786, PMID:22553354], directly represses PKLR to restrain glycolysis and neuroendocrine differentiation in prostate cancer [PMID:35306527], and directly binds and activates the HK1 and ARRDC3 promoters, the latter driving ITGB4 degradation and dampening PI3K/AKT signaling [PMID:37834257, PMID:39873948]. ZBTB10 is also required for NF-κB activation in dendritic cells, where its loss derepresses NKRF and blocks p65/RelB nuclear translocation [PMID:33527393]. Its expression is held in check post-transcriptionally by multiple miRNAs, including miR-27a, miR-361-5p, and miR-582-5p [PMID:21156786, PMID:35608340, PMID:42110628].","teleology":[{"year":2010,"claim":"Established ZBTB10 as a transcriptional repressor of the Sp family and placed it under miRNA control, defining its tumor-suppressive logic of Sp-dependent gene silencing.","evidence":"RNAi knockdown, overexpression, and miR-27a mimic/inhibitor with antioxidant rescue in colon cancer cells","pmids":["21156786"],"confidence":"Medium","gaps":["Direct promoter binding of ZBTB10 at Sp gene loci not demonstrated","Mechanism of repression (corepressor recruitment) not defined"]},{"year":2012,"claim":"Extended the miR-27a/ZBTB10/Sp axis to ErbB2-driven breast cancer and linked it to pharmacological induction, showing the axis is druggable.","evidence":"miR-27a mimic, ZBTB10 overexpression, CB1/CB2 antagonists and Sp RNAi in breast cancer lines","pmids":["22553354"],"confidence":"Medium","gaps":["Direct vs indirect link between CB receptors and ZBTB10 unresolved","No biochemical demonstration of ZBTB10 binding ErbB2 regulatory regions"]},{"year":2019,"claim":"Revealed an unexpected chromatin function by showing ZBTB10 binds the telomeric variant repeat TTGGGG directly and associates with shelterin at ALT telomeres, distinct from its Sp-repressor role.","evidence":"In vitro binding with affinity measurement, ALT-cell co-localization, and reciprocal Co-IP with TRF2/RAP1","pmids":["30629181"],"confidence":"High","gaps":["Functional consequence of telomere binding for ALT maintenance not established","Whether telomeric and transcriptional roles are coupled is unknown"]},{"year":2021,"claim":"Identified a positive regulatory role in innate immunity, showing ZBTB10 is required for NF-κB activation in dendritic cells via suppression of the repressor NKRF.","evidence":"shRNA knockdown in Mutu-DC, transcriptome analysis, p65/RelB translocation blots, and T cell co-culture","pmids":["33527393"],"confidence":"Medium","gaps":["Whether ZBTB10 represses NKRF by direct promoter binding not shown","In vivo dendritic cell requirement not tested"]},{"year":2022,"claim":"Defined ZBTB10 as a direct repressor of PKLR controlling glycolytic and neuroendocrine programs lost after androgen-deprivation therapy in prostate cancer.","evidence":"Loss/gain-of-function with reporter assays in prostate cancer cells","pmids":["35306527"],"confidence":"Medium","gaps":["ChIP-level confirmation of ZBTB10 at the PKLR locus not reported","Mechanism linking AR signaling to ZBTB10 loss unclear"]},{"year":2023,"claim":"Provided the atomic basis of variant-repeat recognition and a single-residue switch (Arg767Gln) that retargets ZBTB10 to canonical telomeric DNA.","evidence":"X-ray crystallography of wild-type and mutant ZF1-2/DNA complexes with ITC validation","pmids":["36657642"],"confidence":"High","gaps":["Biological role of variant-repeat preference in cells not addressed","Structure does not resolve how full-length ZBTB10 engages shelterin"]},{"year":2023,"claim":"Demonstrated ZBTB10 can act as a direct transcriptional activator by binding and activating the HK1 promoter, broadening its activity beyond repression.","evidence":"ChIP and luciferase reporter assays with knockdown/overexpression in laryngeal cancer and 293T cells","pmids":["37834257"],"confidence":"Medium","gaps":["Coactivators mediating activation not identified","Determinants of activator vs repressor mode unknown"]},{"year":2025,"claim":"Connected ZBTB10 transcriptional activation of ARRDC3 to ITGB4 turnover and PI3K/AKT suppression, defining a tumor-suppressive output in gastric cancer.","evidence":"ChIP, Co-IP, reporter assays, xenografts, and phospho-proteomics","pmids":["39873948"],"confidence":"Medium","gaps":["Direct ZBTB10 binding site within ARRDC3 promoter not finely mapped","Whether ARRDC3 is the sole effector unclear"]},{"year":2025,"claim":"Showed exosomal miR-582-5p represses ZBTB10 to derepress TFR1 via H3K27ac, linking ZBTB10 loss to ferroptosis in renal tubular cells.","evidence":"3′UTR dual-luciferase, miRNA overexpression, and H3K27ac ChIP","pmids":["42110628"],"confidence":"Medium","gaps":["How ZBTB10 loss alters TFR1 promoter H3K27ac mechanistically not defined","Direct ZBTB10 occupancy at TFR1 not shown"]},{"year":null,"claim":"How ZBTB10's telomeric variant-repeat binding integrates with its dual transcriptional activator/repressor functions, and what dictates its mode and target selection in different tissues, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying model linking telomere and promoter roles","Cofactor partners distinguishing activation from repression unidentified","In vivo physiological function not established"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[0,1,5,6]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[2,4,5,6]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,5]},{"term_id":"GO:0000228","term_label":"nuclear chromosome","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[2,4,5,6]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[3]}],"complexes":[],"partners":["TRF2","RAP1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q96DT7","full_name":"Zinc finger and BTB domain-containing protein 10","aliases":["Zinc finger protein RIN ZF"],"length_aa":871,"mass_kda":94.9,"function":"May be involved in transcriptional regulation","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/Q96DT7/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ZBTB10","classification":"Not Classified","n_dependent_lines":9,"n_total_lines":1208,"dependency_fraction":0.0074503311258278145},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"IPO8","stoichiometry":10.0},{"gene":"CSNK2B","stoichiometry":0.2},{"gene":"MIF","stoichiometry":0.2},{"gene":"SRP9","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/ZBTB10","total_profiled":1310},"omim":[{"mim_id":"618576","title":"ZINC FINGER- AND BTB DOMAIN-CONTAINING PROTEIN 10; ZBTB10","url":"https://www.omim.org/entry/618576"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoplasm","reliability":"Enhanced"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ZBTB10"},"hgnc":{"alias_symbol":["RINZF","FLJ12752"],"prev_symbol":[]},"alphafold":{"accession":"Q96DT7","domains":[{"cath_id":"3.30.710.10","chopping":"344-397_404-481","consensus_level":"medium","plddt":79.2725,"start":344,"end":481},{"cath_id":"3.30.160.60","chopping":"726-776","consensus_level":"medium","plddt":79.4941,"start":726,"end":776},{"cath_id":"-","chopping":"777-811","consensus_level":"medium","plddt":70.5049,"start":777,"end":811}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96DT7","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q96DT7-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q96DT7-F1-predicted_aligned_error_v6.png","plddt_mean":48.91},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ZBTB10","jax_strain_url":"https://www.jax.org/strain/search?query=ZBTB10"},"sequence":{"accession":"Q96DT7","fasta_url":"https://rest.uniprot.org/uniprotkb/Q96DT7.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q96DT7/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96DT7"}},"corpus_meta":[{"pmid":"21156786","id":"PMC_21156786","title":"GT-094, a NO-NSAID, inhibits colon cancer cell growth by activation of a reactive oxygen species-microRNA-27a: ZBTB10-specificity protein pathway.","date":"2010","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/21156786","citation_count":96,"is_preprint":false},{"pmid":"22407812","id":"PMC_22407812","title":"Betulinic acid decreases ER-negative breast cancer cell growth in vitro and in vivo: role of Sp transcription factors and microRNA-27a:ZBTB10.","date":"2012","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/22407812","citation_count":84,"is_preprint":false},{"pmid":"22553354","id":"PMC_22553354","title":"Betulinic acid targets YY1 and ErbB2 through cannabinoid receptor-dependent disruption of microRNA-27a:ZBTB10 in breast cancer.","date":"2012","source":"Molecular cancer therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/22553354","citation_count":74,"is_preprint":false},{"pmid":"23471840","id":"PMC_23471840","title":"The drug resistance suppression induced by curcuminoids in colon cancer SW-480 cells is mediated by reactive oxygen species-induced disruption of the microRNA-27a-ZBTB10-Sp axis.","date":"2013","source":"Molecular nutrition & food research","url":"https://pubmed.ncbi.nlm.nih.gov/23471840","citation_count":66,"is_preprint":false},{"pmid":"26562150","id":"PMC_26562150","title":"Genome-Wide Association Study of Late-Onset Myasthenia Gravis: Confirmation of TNFRSF11A and Identification of ZBTB10 and Three Distinct HLA Associations.","date":"2015","source":"Molecular medicine (Cambridge, Mass.)","url":"https://pubmed.ncbi.nlm.nih.gov/26562150","citation_count":53,"is_preprint":false},{"pmid":"30629181","id":"PMC_30629181","title":"ZBTB10 binds the telomeric variant repeat TTGGGG and interacts with TRF2.","date":"2019","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/30629181","citation_count":31,"is_preprint":false},{"pmid":"23254909","id":"PMC_23254909","title":"The microRNA-27a: ZBTB10-specificity protein pathway is involved in follicle stimulating hormone-induced VEGF, Cox2 and survivin expression in ovarian epithelial cancer cells.","date":"2012","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/23254909","citation_count":30,"is_preprint":false},{"pmid":"35306527","id":"PMC_35306527","title":"Pyruvate kinase L/R links metabolism dysfunction to neuroendocrine differentiation of prostate cancer by ZBTB10 deficiency.","date":"2022","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/35306527","citation_count":16,"is_preprint":false},{"pmid":"33527393","id":"PMC_33527393","title":"Zbtb10 transcription factor is crucial for murine cDC1 activation and cytokine secretion.","date":"2021","source":"European journal of immunology","url":"https://pubmed.ncbi.nlm.nih.gov/33527393","citation_count":13,"is_preprint":false},{"pmid":"36657642","id":"PMC_36657642","title":"Structural insights into the recognition of telomeric variant repeat TTGGGG by broad-complex, tramtrack and bric-à-brac - zinc finger protein ZBTB10.","date":"2023","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/36657642","citation_count":10,"is_preprint":false},{"pmid":"37834257","id":"PMC_37834257","title":"Intermittent Hypoxia Promotes TAM-Induced Glycolysis in Laryngeal Cancer Cells via Regulation of HK1 Expression through Activation of ZBTB10.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/37834257","citation_count":10,"is_preprint":false},{"pmid":"35608340","id":"PMC_35608340","title":"MiR-361-5p promotes proliferation and inhibits apoptosis of fibroblast-like synoviocytes via targeting ZBTB10 in rheumatoid arthritis.","date":"2022","source":"Autoimmunity","url":"https://pubmed.ncbi.nlm.nih.gov/35608340","citation_count":8,"is_preprint":false},{"pmid":"39873948","id":"PMC_39873948","title":"Enhanced ZBTB10 expression induced by betulinic acid inhibits gastric cancer progression by inactivating the ARRDC3/ITGB4/PI3K/AKT pathway.","date":"2025","source":"Cellular oncology (Dordrecht, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/39873948","citation_count":4,"is_preprint":false},{"pmid":"42110628","id":"PMC_42110628","title":"M1 macrophage exosomes induce ferroptosis via MiR-582-5p-mediated ZBTB10 suppression in sepsis-induced acute kidney injury.","date":"2025","source":"Archives of medical science : AMS","url":"https://pubmed.ncbi.nlm.nih.gov/42110628","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9528,"output_tokens":2653,"usd":0.034189,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9866,"output_tokens":3077,"usd":0.063128,"stage2_stop_reason":"end_turn"},"total_usd":0.097317,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2019,\n      \"finding\": \"ZBTB10 directly binds the telomeric variant repeat sequence TTGGGG via its two C2H2 zinc fingers with nanomolar affinity, co-localizes with a subset of telomeres in ALT-positive U2OS cells, and interacts with TRF2/RAP1 via the N-terminal region of TRF2.\",\n      \"method\": \"In vitro binding assays, co-localization imaging in ALT cells, co-immunoprecipitation with TRF2/RAP1\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — direct binding demonstrated in vitro with affinity measurement, domain mapping, reciprocal Co-IP, and cellular co-localization, multiple orthogonal methods in single study\",\n      \"pmids\": [\"30629181\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Crystal structure of human ZBTB10 ZF1-2 in complex with double-stranded TTGGGG DNA revealed the molecular basis of sequence-specific recognition; a single Arg767Gln substitution shifts preference from TTGGGG to the canonical TTAGGG repeat, as confirmed by co-crystal structure and calorimetric analysis.\",\n      \"method\": \"X-ray crystallography, isothermal titration calorimetry, site-directed mutagenesis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — atomic-resolution crystal structures of wild-type and mutant complexes plus calorimetric validation in one rigorous study\",\n      \"pmids\": [\"36657642\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"ZBTB10 functions as a transcriptional repressor of Sp1, Sp3, and Sp4 in colon cancer cells; its expression is constitutively suppressed by miR-27a, and ROS-mediated downregulation of miR-27a leads to ZBTB10 induction and subsequent repression of Sp-regulated genes (cyclin D1, c-Met, EGFR, bcl-2, survivin, VEGF/VEGFRs).\",\n      \"method\": \"RNA interference knockdown, ZBTB10 expression plasmid transfection, RT-PCR, Western blot, miR-27a mimic/inhibitor experiments, antioxidant rescue (glutathione co-treatment)\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — multiple orthogonal approaches (RNAi, overexpression, mimic/inhibitor, chemical rescue) in single lab across multiple cell lines\",\n      \"pmids\": [\"21156786\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Zbtb10 is required for NF-κB activation in cDC1 dendritic cells; Zbtb10 knockdown enhanced expression of NF-κB repressing factor (NKRF), abrogated p65 and RelB nuclear translocation, and suppressed expression of co-stimulatory molecules CD80/CD86 and cytokines IL-12, IL-6, and IL-10, thereby impairing CD4+ T cell Th1 differentiation.\",\n      \"method\": \"shRNA knockdown in Mutu-DC line, global transcriptome analysis, Western blot for p65/RelB nuclear translocation, T cell co-culture functional assays\",\n      \"journal\": \"European journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KD with defined cellular and molecular phenotype plus mechanistic pathway identification (NKRF/NF-κB), single lab\",\n      \"pmids\": [\"33527393\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ZBTB10 directly transcriptionally represses PKLR expression in prostate cancer cells; loss of ZBTB10 following androgen-deprivation therapy activates PKLR, enhancing glycolysis and neuroendocrine differentiation.\",\n      \"method\": \"Loss-of-function/gain-of-function experiments, RT-PCR, Western blot, transcriptional reporter assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — direct transcriptional regulation demonstrated with overexpression/knockdown and functional readouts in single lab\",\n      \"pmids\": [\"35306527\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ZBTB10 directly binds the HK1 promoter and activates HK1 transcription, as demonstrated by chromatin immunoprecipitation and luciferase reporter assay; ZBTB10 knockdown decreased HK1 expression and ZBTB10 overexpression increased it in laryngeal cancer cells and 293T cells.\",\n      \"method\": \"ChIP assay, luciferase reporter assay, shRNA knockdown, overexpression in cell lines\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct promoter binding and transcriptional activation confirmed by ChIP and reporter assay, single lab\",\n      \"pmids\": [\"37834257\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ZBTB10 directly binds a specific response element in the ARRDC3 promoter to enhance ARRDC3 expression; elevated ARRDC3 interacts with β4-integrin (ITGB4) and promotes its ubiquitination and degradation, leading to reduced PI3K/AKT phosphorylation and suppression of gastric cancer progression.\",\n      \"method\": \"ChIP assay, co-immunoprecipitation, luciferase reporter assay, overexpression/knockdown, xenograft assays, phospho-proteomic profiling\",\n      \"journal\": \"Cellular oncology (Dordrecht, Netherlands)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (ChIP, Co-IP, reporter assay, in vivo xenograft) in single lab establishing direct promoter binding and downstream protein interactions\",\n      \"pmids\": [\"39873948\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"miR-582-5p (delivered via M1 macrophage exosomes) directly binds the 3′UTR of ZBTB10 and suppresses its expression; ZBTB10 suppression leads to increased H3K27ac modification of the TFR1 promoter, enhanced TFR1 transcription, and ferroptosis in renal tubular epithelial cells.\",\n      \"method\": \"Dual-luciferase reporter assay (3′UTR), miR-582-5p overexpression, RT-qPCR, Western blot, ChIP for H3K27ac\",\n      \"journal\": \"Archives of medical science : AMS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — 3′UTR luciferase validation plus H3K27ac ChIP linking ZBTB10 loss to TFR1 epigenetic regulation, single lab with multiple methods\",\n      \"pmids\": [\"42110628\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"miR-27a constitutively inhibits ZBTB10 expression; in ErbB2-overexpressing breast cancer cells, betulinic acid induces ZBTB10 and represses Sp1/Sp3/Sp4 and their downstream target ErbB2 (via YY1) through cannabinoid receptor CB1 and CB2-dependent mechanisms.\",\n      \"method\": \"miR-27a mimic transfection, ZBTB10 overexpression, CB1/CB2 receptor antagonists, RNA interference of Sp1/Sp3/Sp4\",\n      \"journal\": \"Molecular cancer therapeutics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — receptor-dependency established pharmacologically with multiple cell lines and RNAi rescue, single lab\",\n      \"pmids\": [\"22553354\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"miR-361-5p directly targets the 3′UTR of ZBTB10 (confirmed by dual luciferase assay) and suppresses its expression; ZBTB10 overexpression reverses the pro-proliferative, anti-apoptotic effects of miR-361-5p in rheumatoid arthritis fibroblast-like synoviocytes.\",\n      \"method\": \"Dual luciferase reporter assay, ZBTB10 overexpression rescue experiments, flow cytometry, Western blot\",\n      \"journal\": \"Autoimmunity\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — 3′UTR luciferase validation plus rescue experiment, single lab\",\n      \"pmids\": [\"35608340\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ZBTB10 is a BTB-ZF transcription factor that uses its tandem C2H2 zinc fingers to bind the telomeric variant repeat TTGGGG with nanomolar affinity (crystal structure resolved) and interacts with TRF2/RAP1 at ALT telomeres; in the nucleus it directly represses Sp1/Sp3/Sp4 transcription factors (with downstream effects on VEGF, survivin, ErbB2, and MDR1), directly activates target gene promoters (HK1, ARRDC3), and transcriptionally represses PKLR to control glycolytic and neuroendocrine programs, while its own expression is post-transcriptionally suppressed by multiple miRNAs (miR-27a, miR-361-5p, miR-582-5p) and it supports NF-κB-dependent activation of dendritic cells.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ZBTB10 is a BTB-ZF transcription factor that uses tandem C2H2 zinc fingers to read specific DNA sequences and to regulate transcriptional programs across telomere biology, cancer metabolism, and immune cell activation [#0, #2]. At telomeres, ZBTB10 binds the variant repeat TTGGGG with nanomolar affinity, co-localizes with a subset of telomeres in ALT-positive cells, and associates with the shelterin proteins TRF2/RAP1; a crystal structure of its ZF1-2 domain bound to TTGGGG defines the basis of sequence-specific recognition, where a single Arg767Gln substitution redirects preference toward the canonical TTAGGG repeat [#0, #1]. As a transcriptional regulator it acts bidirectionally: it represses the Sp transcription factor family (Sp1/Sp3/Sp4) and their downstream targets including survivin, VEGF, and ErbB2 [#2, #8], directly represses PKLR to restrain glycolysis and neuroendocrine differentiation in prostate cancer [#4], and directly binds and activates the HK1 and ARRDC3 promoters, the latter driving ITGB4 degradation and dampening PI3K/AKT signaling [#5, #6]. ZBTB10 is also required for NF-\\u03baB activation in dendritic cells, where its loss derepresses NKRF and blocks p65/RelB nuclear translocation [#3]. Its expression is held in check post-transcriptionally by multiple miRNAs, including miR-27a, miR-361-5p, and miR-582-5p [#2, #9, #7].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Established ZBTB10 as a transcriptional repressor of the Sp family and placed it under miRNA control, defining its tumor-suppressive logic of Sp-dependent gene silencing.\",\n      \"evidence\": \"RNAi knockdown, overexpression, and miR-27a mimic/inhibitor with antioxidant rescue in colon cancer cells\",\n      \"pmids\": [\"21156786\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct promoter binding of ZBTB10 at Sp gene loci not demonstrated\", \"Mechanism of repression (corepressor recruitment) not defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Extended the miR-27a/ZBTB10/Sp axis to ErbB2-driven breast cancer and linked it to pharmacological induction, showing the axis is druggable.\",\n      \"evidence\": \"miR-27a mimic, ZBTB10 overexpression, CB1/CB2 antagonists and Sp RNAi in breast cancer lines\",\n      \"pmids\": [\"22553354\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect link between CB receptors and ZBTB10 unresolved\", \"No biochemical demonstration of ZBTB10 binding ErbB2 regulatory regions\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Revealed an unexpected chromatin function by showing ZBTB10 binds the telomeric variant repeat TTGGGG directly and associates with shelterin at ALT telomeres, distinct from its Sp-repressor role.\",\n      \"evidence\": \"In vitro binding with affinity measurement, ALT-cell co-localization, and reciprocal Co-IP with TRF2/RAP1\",\n      \"pmids\": [\"30629181\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of telomere binding for ALT maintenance not established\", \"Whether telomeric and transcriptional roles are coupled is unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified a positive regulatory role in innate immunity, showing ZBTB10 is required for NF-\\u03baB activation in dendritic cells via suppression of the repressor NKRF.\",\n      \"evidence\": \"shRNA knockdown in Mutu-DC, transcriptome analysis, p65/RelB translocation blots, and T cell co-culture\",\n      \"pmids\": [\"33527393\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ZBTB10 represses NKRF by direct promoter binding not shown\", \"In vivo dendritic cell requirement not tested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined ZBTB10 as a direct repressor of PKLR controlling glycolytic and neuroendocrine programs lost after androgen-deprivation therapy in prostate cancer.\",\n      \"evidence\": \"Loss/gain-of-function with reporter assays in prostate cancer cells\",\n      \"pmids\": [\"35306527\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ChIP-level confirmation of ZBTB10 at the PKLR locus not reported\", \"Mechanism linking AR signaling to ZBTB10 loss unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Provided the atomic basis of variant-repeat recognition and a single-residue switch (Arg767Gln) that retargets ZBTB10 to canonical telomeric DNA.\",\n      \"evidence\": \"X-ray crystallography of wild-type and mutant ZF1-2/DNA complexes with ITC validation\",\n      \"pmids\": [\"36657642\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biological role of variant-repeat preference in cells not addressed\", \"Structure does not resolve how full-length ZBTB10 engages shelterin\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrated ZBTB10 can act as a direct transcriptional activator by binding and activating the HK1 promoter, broadening its activity beyond repression.\",\n      \"evidence\": \"ChIP and luciferase reporter assays with knockdown/overexpression in laryngeal cancer and 293T cells\",\n      \"pmids\": [\"37834257\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Coactivators mediating activation not identified\", \"Determinants of activator vs repressor mode unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected ZBTB10 transcriptional activation of ARRDC3 to ITGB4 turnover and PI3K/AKT suppression, defining a tumor-suppressive output in gastric cancer.\",\n      \"evidence\": \"ChIP, Co-IP, reporter assays, xenografts, and phospho-proteomics\",\n      \"pmids\": [\"39873948\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ZBTB10 binding site within ARRDC3 promoter not finely mapped\", \"Whether ARRDC3 is the sole effector unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showed exosomal miR-582-5p represses ZBTB10 to derepress TFR1 via H3K27ac, linking ZBTB10 loss to ferroptosis in renal tubular cells.\",\n      \"evidence\": \"3\\u2032UTR dual-luciferase, miRNA overexpression, and H3K27ac ChIP\",\n      \"pmids\": [\"42110628\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How ZBTB10 loss alters TFR1 promoter H3K27ac mechanistically not defined\", \"Direct ZBTB10 occupancy at TFR1 not shown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How ZBTB10's telomeric variant-repeat binding integrates with its dual transcriptional activator/repressor functions, and what dictates its mode and target selection in different tissues, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying model linking telomere and promoter roles\", \"Cofactor partners distinguishing activation from repression unidentified\", \"In vivo physiological function not established\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [0, 1, 5, 6]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [2, 4, 5, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 5]},\n      {\"term_id\": \"GO:0000228\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [2, 4, 5, 6]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TRF2\", \"RAP1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":5,"faith_total":5,"faith_pct":100.0}}