{"gene":"ZNF471","run_date":"2026-06-11T09:02:07","timeline":{"discoveries":[{"year":2018,"finding":"ZNF471 directly binds to the promoters of TFAP2A and PLS3 (demonstrated by ChIP-PCR) and transcriptionally represses their expression; ZNF471 recruits co-repressor KAP1 to these promoter regions, inducing H3K9me3 enrichment for heterochromatic transcriptional silencing.","method":"ChIP-PCR, ectopic expression, Western blot, bioinformatics modeling","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal ChIP-PCR demonstrating direct promoter binding and H3K9me3 enrichment, with KAP1 co-recruitment, validated by ectopic expression functional assays in multiple cell lines","pmids":["29610526"],"is_preprint":false},{"year":2020,"finding":"ZNF471 directly binds to the MAPK10/JNK3 promoter and activates MAPK10/JNK3 expression and downstream signaling, as well as activating PCDH family gene expression, in esophageal squamous cell carcinoma cells.","method":"ChIP assay (promoter binding), RT-PCR, ectopic expression, in vitro and in vivo functional assays","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct promoter binding shown by ChIP, functional rescue confirmed in vitro and in vivo, single lab","pmids":["32089740"],"is_preprint":false},{"year":2020,"finding":"ZNF471 exerts tumor-suppressive functions in breast cancer cells through suppression of AKT and Wnt/β-catenin signaling pathways, inhibiting epithelial-mesenchymal transition and tumor cell stemness.","method":"Western blot, CCK-8, clonogenicity, wound healing, Transwell, flow cytometry, nude mice tumorigenicity, BrdU-ELISA, knockdown and overexpression","journal":"Clinical epigenetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — defined pathway suppression (AKT, Wnt/β-catenin) confirmed by Western blot with KD and OE, multiple orthogonal cellular assays, single lab","pmids":["33203470"],"is_preprint":false},{"year":2024,"finding":"ZNF471 physically interacts with BANP (demonstrated by co-immunoprecipitation or pull-down) and suppresses the malignant phenotype of renal cell carcinoma by inactivating the PI3K/AKT/mTOR signaling pathway.","method":"Transcriptome sequencing, bioinformatics analysis, molecular biology experiments (co-IP/interaction assay), cell biology assays","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — protein-protein interaction with BANP and pathway inactivation (PI3K/AKT/mTOR) reported from single lab with molecular biology experiments; abstract does not detail orthogonal validation","pmids":["38169650"],"is_preprint":false},{"year":2024,"finding":"Asparagine 358 is a functional N-glycosylation site on ZNF471; suppression of N-glycosylation at this site enhances ZNF471 protein stability and promotes its translocation to the cell nucleus, where it binds the c-Myc promoter to repress c-Myc expression and thereby exerts anti-cancer effects and increases docetaxel sensitivity in tongue squamous cell carcinoma.","method":"Site-specific mutagenesis, Western blot, qRT-PCR, immunohistochemistry, nuclear/cytoplasmic fractionation, ChIP (promoter binding)","journal":"The American journal of pathology","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — site-specific mutagenesis identifying glycosylation site combined with functional nuclear translocation and promoter-binding assay; single lab","pmids":["38749608"],"is_preprint":false},{"year":2025,"finding":"ZNF471 overexpression reduces protein levels of β-catenin and downstream Wnt/β-catenin targets c-Myc and MMP-7, inhibiting NPC cell growth, migration, invasion, and tumor sphere formation.","method":"Western blot, CCK-8, EdU proliferation assay, wound healing, Transwell invasion assay, spheroid formation assay, overexpression in NPC cell lines","journal":"General physiology and biophysics","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — pathway suppression shown by Western blot with overexpression, multiple phenotypic readouts, single lab, single method per endpoint","pmids":["40326975"],"is_preprint":false},{"year":2026,"finding":"ZNF471 physically interacts with MIS18A and suppresses hepatocellular carcinoma proliferation and metastasis through negative regulation of the Wnt/β-catenin signaling pathway and inhibition of EMT; the ZNF471-MIS18A axis also increases HCC cell sensitivity to apatinib and donafenib.","method":"Transcriptome sequencing, molecular biology experiments (co-IP/interaction assay), Western blot, functional cellular assays, clinical tissue samples","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — protein-protein interaction with MIS18A and Wnt/β-catenin pathway modulation reported from single lab; abstract lacks explicit orthogonal validation of interaction","pmids":["42155792"],"is_preprint":false},{"year":2020,"finding":"In non-small cell lung cancer, ZNF471 acts as a downstream target of miR-942-5p; lncRNA LIFR-AS1 sponges miR-942-5p to derepress ZNF471, and ZNF471 overexpression inhibits NSCLC cell invasion and migration.","method":"Luciferase reporter assay (sponge relationship), overexpression/knockdown of LIFR-AS1 and ZNF471, rescue experiments, in vivo lung metastasis assay","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis/rescue experiments placing ZNF471 downstream of LIFR-AS1/miR-942-5p axis, in vivo confirmation, single lab","pmids":["32489316"],"is_preprint":false}],"current_model":"ZNF471 is a KRAB-domain zinc finger transcriptional repressor that binds target gene promoters (including TFAP2A, PLS3, MAPK10/JNK3, and c-Myc) via its zinc finger motifs, recruits the co-repressor KAP1 to induce H3K9me3-dependent heterochromatin and transcriptional silencing, and also forms complexes with BANP and MIS18A; collectively, these activities suppress AKT, PI3K/AKT/mTOR, and Wnt/β-catenin signaling to inhibit cancer cell proliferation, invasion, and EMT, while its N-glycosylation at Asn358 regulates its protein stability and nuclear translocation; ZNF471 is frequently silenced across multiple tumor types by promoter CpG hypermethylation."},"narrative":{"mechanistic_narrative":"ZNF471 is a KRAB-domain zinc finger transcription factor that functions as a tumor suppressor across multiple epithelial cancers by acting as a sequence-specific regulator of growth- and motility-associated promoters [PMID:29610526, PMID:33203470]. As a transcriptional repressor it binds target promoters such as TFAP2A and PLS3 and recruits the co-repressor KAP1 to nucleate H3K9me3-marked heterochromatin and silence transcription [PMID:29610526]; it also directly represses c-Myc by binding its promoter [PMID:38749608]. In a context-dependent manner ZNF471 can instead activate transcription, binding the MAPK10/JNK3 promoter to induce JNK3 and PCDH family expression in esophageal squamous carcinoma [PMID:32089740]. Downstream of these transcriptional activities, ZNF471 suppresses AKT, PI3K/AKT/mTOR, and Wnt/β-catenin signaling, thereby inhibiting proliferation, invasion, epithelial-mesenchymal transition, and tumor cell stemness [PMID:33203470, PMID:38169650, PMID:40326975]. Its activity is gated by post-translational and physical regulation: N-glycosylation at Asn358 destabilizes the protein and limits its nuclear translocation, so loss of glycosylation enhances stability and nuclear repressor function [PMID:38749608], and it forms complexes with BANP and MIS18A that contribute to pathway suppression [PMID:38169650, PMID:42155792]. Expression of ZNF471 is itself controlled in trans, being derepressed by the LIFR-AS1/miR-942-5p axis in lung cancer [PMID:32489316].","teleology":[{"year":2018,"claim":"Established the core molecular mechanism by showing ZNF471 is a KRAB-zinc-finger repressor that binds specific promoters and recruits KAP1 to deposit repressive H3K9me3, defining how it silences target genes.","evidence":"ChIP-PCR, ectopic expression and Western blot at TFAP2A and PLS3 promoters in cancer cell lines","pmids":["29610526"],"confidence":"High","gaps":["Full target gene repertoire beyond TFAP2A and PLS3 not defined","Direct demonstration of KRAB domain requirement for KAP1 recruitment not shown","No structural basis for promoter recognition"]},{"year":2020,"claim":"Revealed that ZNF471 can also act as a transcriptional activator, binding the MAPK10/JNK3 promoter to induce JNK3 and PCDH expression, broadening its role beyond pure repression.","evidence":"ChIP, RT-PCR and in vitro/in vivo functional assays in esophageal squamous cell carcinoma","pmids":["32089740"],"confidence":"Medium","gaps":["Mechanism distinguishing activating from repressive promoter engagement unknown","No co-activator identified","Single lab"]},{"year":2020,"claim":"Connected ZNF471 transcriptional activity to downstream oncogenic signaling, showing suppression of AKT and Wnt/β-catenin pathways inhibits EMT and stemness, and placed ZNF471 in a regulatory ncRNA axis.","evidence":"Knockdown/overexpression with Western blot and orthogonal cellular assays in breast cancer; luciferase sponge and rescue assays defining the LIFR-AS1/miR-942-5p axis in NSCLC","pmids":["33203470","32489316"],"confidence":"Medium","gaps":["Direct transcriptional targets linking ZNF471 to AKT/Wnt not identified","Whether signaling suppression is direct or secondary to gene silencing unresolved"]},{"year":2024,"claim":"Identified physical protein partners and a post-translational control point, showing ZNF471 interacts with BANP and that Asn358 N-glycosylation regulates its stability and nuclear localization for c-Myc repression.","evidence":"Co-IP/pull-down with BANP and PI3K/AKT/mTOR readouts in RCC; site-specific mutagenesis, fractionation and ChIP at the c-Myc promoter in tongue squamous carcinoma","pmids":["38169650","38749608"],"confidence":"Medium","gaps":["BANP interaction lacks reciprocal/orthogonal validation in abstract","Enzyme mediating Asn358 glycosylation unknown","Functional consequence of BANP binding on transcription not mapped"]},{"year":2026,"claim":"Extended the partner network and therapeutic relevance by defining a ZNF471-MIS18A axis that suppresses Wnt/β-catenin signaling and EMT and modulates drug sensitivity in HCC.","evidence":"Transcriptome sequencing, co-IP, Western blot and functional assays with clinical tissue in hepatocellular carcinoma","pmids":["42155792"],"confidence":"Medium","gaps":["Interaction lacks explicit orthogonal validation","How MIS18A binding alters ZNF471 transcriptional output unknown"]},{"year":null,"claim":"How ZNF471 switches between repressive and activating modes, and how its physical partners (KAP1, BANP, MIS18A) coordinate on chromatin, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No genome-wide binding map","No structural model of zinc finger-DNA recognition","Determinants of activation versus repression undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[0,1,4]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[0,1,4]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0,1,4]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,3,5,6]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[0]}],"complexes":[],"partners":["KAP1","BANP","MIS18A"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BX82","full_name":"Zinc finger protein 471","aliases":["EZFIT-related protein 1"],"length_aa":626,"mass_kda":73.0,"function":"May be involved in transcriptional regulation","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9BX82/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ZNF471","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ZNF471","total_profiled":1310},"omim":[{"mim_id":"620162","title":"ZINC FINGER PROTEIN 471; ZNF471","url":"https://www.omim.org/entry/620162"},{"mim_id":"609451","title":"ZFP90 ZINC FINGER PROTEIN; ZFP90","url":"https://www.omim.org/entry/609451"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ZNF471"},"hgnc":{"alias_symbol":["KIAA1396","Z1971","Zfp78"],"prev_symbol":[]},"alphafold":{"accession":"Q9BX82","domains":[{"cath_id":"-","chopping":"15-67","consensus_level":"high","plddt":79.1913,"start":15,"end":67},{"cath_id":"3.30.160.60","chopping":"514-621","consensus_level":"medium","plddt":87.3196,"start":514,"end":621}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BX82","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BX82-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BX82-F1-predicted_aligned_error_v6.png","plddt_mean":72.81},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ZNF471","jax_strain_url":"https://www.jax.org/strain/search?query=ZNF471"},"sequence":{"accession":"Q9BX82","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BX82.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BX82/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BX82"}},"corpus_meta":[{"pmid":"33672287","id":"PMC_33672287","title":"KRAB-ZFP Transcriptional Regulators Acting as Oncogenes and Tumor Suppressors: An Overview.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33672287","citation_count":69,"is_preprint":false},{"pmid":"32089740","id":"PMC_32089740","title":"19q13 KRAB zinc-finger protein ZNF471 activates MAPK10/JNK3 signaling but is frequently silenced by promoter CpG methylation in esophageal cancer.","date":"2020","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/32089740","citation_count":49,"is_preprint":false},{"pmid":"29610526","id":"PMC_29610526","title":"Zinc-finger protein 471 suppresses gastric cancer through transcriptionally repressing downstream oncogenic PLS3 and TFAP2A.","date":"2018","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/29610526","citation_count":48,"is_preprint":false},{"pmid":"32489316","id":"PMC_32489316","title":"lncRNA LIFR-AS1 suppresses invasion and metastasis of non-small cell lung cancer via the miR-942-5p/ZNF471 axis.","date":"2020","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/32489316","citation_count":43,"is_preprint":false},{"pmid":"33203470","id":"PMC_33203470","title":"The tumor suppressor Zinc finger protein 471 suppresses breast cancer growth and metastasis through inhibiting AKT and Wnt/β-catenin signaling.","date":"2020","source":"Clinical epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/33203470","citation_count":29,"is_preprint":false},{"pmid":"28255813","id":"PMC_28255813","title":"Gene promoter-associated CpG island hypermethylation in squamous cell carcinoma of the tongue.","date":"2017","source":"Virchows Archiv : an international journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/28255813","citation_count":26,"is_preprint":false},{"pmid":"34944472","id":"PMC_34944472","title":"Epigenome-Wide Association Study of Prostate Cancer in African Americans Identifies DNA Methylation Biomarkers for Aggressive Disease.","date":"2021","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/34944472","citation_count":13,"is_preprint":false},{"pmid":"38169650","id":"PMC_38169650","title":"ZNF471 Interacts with BANP to Reduce Tumour Malignancy by Inactivating PI3K/AKT/mTOR Signalling but is Frequently Silenced by Aberrant Promoter Methylation in Renal Cell Carcinoma.","date":"2024","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38169650","citation_count":10,"is_preprint":false},{"pmid":"36245370","id":"PMC_36245370","title":"Salivary DNA methylation markers for cancer of oral cavity.","date":"2022","source":"Cancer biomarkers : section A of Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/36245370","citation_count":6,"is_preprint":false},{"pmid":"31098739","id":"PMC_31098739","title":"Colla corii asini might upregulate ZNF471 and THOC5 by KRAB domain-containing zinc-finger protein pathway and THO complex subunit 5 pathway to improve anemia of pregnant women with β-thalassemia.","date":"2019","source":"Annals of hematology","url":"https://pubmed.ncbi.nlm.nih.gov/31098739","citation_count":5,"is_preprint":false},{"pmid":"38749608","id":"PMC_38749608","title":"Suppression of N-Glycosylation of Zinc Finger Protein 471 Affects Proliferation, Invasion, and Docetaxel Sensitivity of Tongue Squamous Cell Carcinoma via Regulation of c-Myc.","date":"2024","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/38749608","citation_count":5,"is_preprint":false},{"pmid":"37779184","id":"PMC_37779184","title":"Screening and identifying of biomarkers in early colorectal cancer and adenoma based on genome-wide methylation profiles.","date":"2023","source":"World journal of surgical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/37779184","citation_count":4,"is_preprint":false},{"pmid":"40326975","id":"PMC_40326975","title":"ZNF471 inhibits nasopharyngeal carcinoma cell growth and stemness.","date":"2025","source":"General physiology and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/40326975","citation_count":1,"is_preprint":false},{"pmid":"42155792","id":"PMC_42155792","title":"Methylation of zinc-finger protein ZNF471 regulates MIS18A expression and inhibits proliferation and metastasis of hepatocellular carcinoma via negative regulation of the Wnt/β-catenin pathway, as well as enhances sensitivity to apatinib and donafenib.","date":"2026","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/42155792","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10079,"output_tokens":2076,"usd":0.030689,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9057,"output_tokens":2361,"usd":0.052155,"stage2_stop_reason":"end_turn"},"total_usd":0.082844,"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\": 2018,\n      \"finding\": \"ZNF471 directly binds to the promoters of TFAP2A and PLS3 (demonstrated by ChIP-PCR) and transcriptionally represses their expression; ZNF471 recruits co-repressor KAP1 to these promoter regions, inducing H3K9me3 enrichment for heterochromatic transcriptional silencing.\",\n      \"method\": \"ChIP-PCR, ectopic expression, Western blot, bioinformatics modeling\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal ChIP-PCR demonstrating direct promoter binding and H3K9me3 enrichment, with KAP1 co-recruitment, validated by ectopic expression functional assays in multiple cell lines\",\n      \"pmids\": [\"29610526\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ZNF471 directly binds to the MAPK10/JNK3 promoter and activates MAPK10/JNK3 expression and downstream signaling, as well as activating PCDH family gene expression, in esophageal squamous cell carcinoma cells.\",\n      \"method\": \"ChIP assay (promoter binding), RT-PCR, ectopic expression, in vitro and in vivo functional assays\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct promoter binding shown by ChIP, functional rescue confirmed in vitro and in vivo, single lab\",\n      \"pmids\": [\"32089740\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ZNF471 exerts tumor-suppressive functions in breast cancer cells through suppression of AKT and Wnt/β-catenin signaling pathways, inhibiting epithelial-mesenchymal transition and tumor cell stemness.\",\n      \"method\": \"Western blot, CCK-8, clonogenicity, wound healing, Transwell, flow cytometry, nude mice tumorigenicity, BrdU-ELISA, knockdown and overexpression\",\n      \"journal\": \"Clinical epigenetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — defined pathway suppression (AKT, Wnt/β-catenin) confirmed by Western blot with KD and OE, multiple orthogonal cellular assays, single lab\",\n      \"pmids\": [\"33203470\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZNF471 physically interacts with BANP (demonstrated by co-immunoprecipitation or pull-down) and suppresses the malignant phenotype of renal cell carcinoma by inactivating the PI3K/AKT/mTOR signaling pathway.\",\n      \"method\": \"Transcriptome sequencing, bioinformatics analysis, molecular biology experiments (co-IP/interaction assay), cell biology assays\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — protein-protein interaction with BANP and pathway inactivation (PI3K/AKT/mTOR) reported from single lab with molecular biology experiments; abstract does not detail orthogonal validation\",\n      \"pmids\": [\"38169650\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Asparagine 358 is a functional N-glycosylation site on ZNF471; suppression of N-glycosylation at this site enhances ZNF471 protein stability and promotes its translocation to the cell nucleus, where it binds the c-Myc promoter to repress c-Myc expression and thereby exerts anti-cancer effects and increases docetaxel sensitivity in tongue squamous cell carcinoma.\",\n      \"method\": \"Site-specific mutagenesis, Western blot, qRT-PCR, immunohistochemistry, nuclear/cytoplasmic fractionation, ChIP (promoter binding)\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — site-specific mutagenesis identifying glycosylation site combined with functional nuclear translocation and promoter-binding assay; single lab\",\n      \"pmids\": [\"38749608\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ZNF471 overexpression reduces protein levels of β-catenin and downstream Wnt/β-catenin targets c-Myc and MMP-7, inhibiting NPC cell growth, migration, invasion, and tumor sphere formation.\",\n      \"method\": \"Western blot, CCK-8, EdU proliferation assay, wound healing, Transwell invasion assay, spheroid formation assay, overexpression in NPC cell lines\",\n      \"journal\": \"General physiology and biophysics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — pathway suppression shown by Western blot with overexpression, multiple phenotypic readouts, single lab, single method per endpoint\",\n      \"pmids\": [\"40326975\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"ZNF471 physically interacts with MIS18A and suppresses hepatocellular carcinoma proliferation and metastasis through negative regulation of the Wnt/β-catenin signaling pathway and inhibition of EMT; the ZNF471-MIS18A axis also increases HCC cell sensitivity to apatinib and donafenib.\",\n      \"method\": \"Transcriptome sequencing, molecular biology experiments (co-IP/interaction assay), Western blot, functional cellular assays, clinical tissue samples\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — protein-protein interaction with MIS18A and Wnt/β-catenin pathway modulation reported from single lab; abstract lacks explicit orthogonal validation of interaction\",\n      \"pmids\": [\"42155792\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In non-small cell lung cancer, ZNF471 acts as a downstream target of miR-942-5p; lncRNA LIFR-AS1 sponges miR-942-5p to derepress ZNF471, and ZNF471 overexpression inhibits NSCLC cell invasion and migration.\",\n      \"method\": \"Luciferase reporter assay (sponge relationship), overexpression/knockdown of LIFR-AS1 and ZNF471, rescue experiments, in vivo lung metastasis assay\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis/rescue experiments placing ZNF471 downstream of LIFR-AS1/miR-942-5p axis, in vivo confirmation, single lab\",\n      \"pmids\": [\"32489316\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ZNF471 is a KRAB-domain zinc finger transcriptional repressor that binds target gene promoters (including TFAP2A, PLS3, MAPK10/JNK3, and c-Myc) via its zinc finger motifs, recruits the co-repressor KAP1 to induce H3K9me3-dependent heterochromatin and transcriptional silencing, and also forms complexes with BANP and MIS18A; collectively, these activities suppress AKT, PI3K/AKT/mTOR, and Wnt/β-catenin signaling to inhibit cancer cell proliferation, invasion, and EMT, while its N-glycosylation at Asn358 regulates its protein stability and nuclear translocation; ZNF471 is frequently silenced across multiple tumor types by promoter CpG hypermethylation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ZNF471 is a KRAB-domain zinc finger transcription factor that functions as a tumor suppressor across multiple epithelial cancers by acting as a sequence-specific regulator of growth- and motility-associated promoters [#0, #2]. As a transcriptional repressor it binds target promoters such as TFAP2A and PLS3 and recruits the co-repressor KAP1 to nucleate H3K9me3-marked heterochromatin and silence transcription [#0]; it also directly represses c-Myc by binding its promoter [#4]. In a context-dependent manner ZNF471 can instead activate transcription, binding the MAPK10/JNK3 promoter to induce JNK3 and PCDH family expression in esophageal squamous carcinoma [#1]. Downstream of these transcriptional activities, ZNF471 suppresses AKT, PI3K/AKT/mTOR, and Wnt/\\u03b2-catenin signaling, thereby inhibiting proliferation, invasion, epithelial-mesenchymal transition, and tumor cell stemness [#2, #3, #5]. Its activity is gated by post-translational and physical regulation: N-glycosylation at Asn358 destabilizes the protein and limits its nuclear translocation, so loss of glycosylation enhances stability and nuclear repressor function [#4], and it forms complexes with BANP and MIS18A that contribute to pathway suppression [#3, #6]. Expression of ZNF471 is itself controlled in trans, being derepressed by the LIFR-AS1/miR-942-5p axis in lung cancer [#7].\",\n  \"teleology\": [\n    {\n      \"year\": 2018,\n      \"claim\": \"Established the core molecular mechanism by showing ZNF471 is a KRAB-zinc-finger repressor that binds specific promoters and recruits KAP1 to deposit repressive H3K9me3, defining how it silences target genes.\",\n      \"evidence\": \"ChIP-PCR, ectopic expression and Western blot at TFAP2A and PLS3 promoters in cancer cell lines\",\n      \"pmids\": [\"29610526\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full target gene repertoire beyond TFAP2A and PLS3 not defined\", \"Direct demonstration of KRAB domain requirement for KAP1 recruitment not shown\", \"No structural basis for promoter recognition\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Revealed that ZNF471 can also act as a transcriptional activator, binding the MAPK10/JNK3 promoter to induce JNK3 and PCDH expression, broadening its role beyond pure repression.\",\n      \"evidence\": \"ChIP, RT-PCR and in vitro/in vivo functional assays in esophageal squamous cell carcinoma\",\n      \"pmids\": [\"32089740\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism distinguishing activating from repressive promoter engagement unknown\", \"No co-activator identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected ZNF471 transcriptional activity to downstream oncogenic signaling, showing suppression of AKT and Wnt/\\u03b2-catenin pathways inhibits EMT and stemness, and placed ZNF471 in a regulatory ncRNA axis.\",\n      \"evidence\": \"Knockdown/overexpression with Western blot and orthogonal cellular assays in breast cancer; luciferase sponge and rescue assays defining the LIFR-AS1/miR-942-5p axis in NSCLC\",\n      \"pmids\": [\"33203470\", \"32489316\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct transcriptional targets linking ZNF471 to AKT/Wnt not identified\", \"Whether signaling suppression is direct or secondary to gene silencing unresolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified physical protein partners and a post-translational control point, showing ZNF471 interacts with BANP and that Asn358 N-glycosylation regulates its stability and nuclear localization for c-Myc repression.\",\n      \"evidence\": \"Co-IP/pull-down with BANP and PI3K/AKT/mTOR readouts in RCC; site-specific mutagenesis, fractionation and ChIP at the c-Myc promoter in tongue squamous carcinoma\",\n      \"pmids\": [\"38169650\", \"38749608\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"BANP interaction lacks reciprocal/orthogonal validation in abstract\", \"Enzyme mediating Asn358 glycosylation unknown\", \"Functional consequence of BANP binding on transcription not mapped\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Extended the partner network and therapeutic relevance by defining a ZNF471-MIS18A axis that suppresses Wnt/\\u03b2-catenin signaling and EMT and modulates drug sensitivity in HCC.\",\n      \"evidence\": \"Transcriptome sequencing, co-IP, Western blot and functional assays with clinical tissue in hepatocellular carcinoma\",\n      \"pmids\": [\"42155792\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interaction lacks explicit orthogonal validation\", \"How MIS18A binding alters ZNF471 transcriptional output unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How ZNF471 switches between repressive and activating modes, and how its physical partners (KAP1, BANP, MIS18A) coordinate on chromatin, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No genome-wide binding map\", \"No structural model of zinc finger-DNA recognition\", \"Determinants of activation versus repression undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 1, 4]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [0, 1, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0, 1, 4]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 3, 5, 6]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"KAP1\", \"BANP\", \"MIS18A\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":6,"faith_total":6,"faith_pct":100.0}}