{"gene":"ANKRD6","run_date":"2026-04-28T17:12:37","timeline":{"discoveries":[{"year":2014,"finding":"mAnkrd6 protein is asymmetrically localized in cells of the mouse inner ear sensory organs, characteristic of core PCP complex components. Loss of mAnkrd6 causes planar cell polarity defects in inner ear sensory organs, and canonical Wnt signaling is significantly increased in mouse embryonic fibroblasts from mAnkrd6 knockout mice compared to wild-type controls, demonstrating that mAnkrd6 suppresses canonical Wnt signaling.","method":"Mouse knockout (loss-of-function), immunolocalization, canonical Wnt reporter assays in MEFs, Drosophila rescue experiments","journal":"Developmental biology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (KO phenotype, localization, reporter assays, cross-species rescue) in a single study","pmids":["25218921"],"is_preprint":false},{"year":2014,"finding":"ANKRD6 (Diversin) acts as a molecular switch between canonical Wnt/β-catenin and non-canonical PCP Wnt signaling pathways; rare missense mutations p.Pro548Leu and p.Arg632His identified in NTD patients significantly altered DIVERSIN activity in Wnt signaling reporter assays in a hypomorphic manner.","method":"Wnt signaling reporter assays in mammalian cells, sequencing of coding region in NTD patients vs. controls","journal":"Birth defects research. Part A, Clinical and molecular teratology","confidence":"Medium","confidence_rationale":"Tier 2 — functional reporter assay with specific mutation validation, single lab","pmids":["25200652"],"is_preprint":false},{"year":2005,"finding":"ANKRD6 ankyrin repeats are predicted binding domains for Prickle1, Prickle2, Vangl1, and Vangl2; the central coiled-coil region is located within the binding domain for Casein kinase Iε (CKIε); and the C-terminal coiled-coil region is located within the binding domain for Axin1 and Axin2, placing ANKRD6 in direct physical interaction with core PCP and canonical Wnt pathway components.","method":"Bioinformatics domain analysis corroborated by literature on known interaction domains","journal":"International journal of molecular medicine","confidence":"Low","confidence_rationale":"Tier 4 — computational/bioinformatics prediction only, no direct binding experiment in this paper","pmids":["15647854"],"is_preprint":false},{"year":2005,"finding":"ANKRD6 (Diego homolog) interacts with PRICKLE1, PRICKLE2, VANGL1, VANGL2, DVL1, DVL2, DVL3 as part of the vertebrate PCP signaling complex, and functions to induce class switch from the WNT/GSK3β (canonical) pathway to the WNT/PCP pathway.","method":"Comparative genomics and literature synthesis identifying protein-protein interactions","journal":"International journal of oncology","confidence":"Low","confidence_rationale":"Tier 4 — bioinformatics/comparative genomics, interactions not directly assayed in this paper","pmids":["15809738"],"is_preprint":false},{"year":2019,"finding":"miR-214 targets ANKRD6 in melanoma cells; knockdown of ANKRD6 increased melanoma cell proliferation and migration and decreased sensitivity to MAPK inhibitors, indicating ANKRD6 functions as a negative regulator of Wnt/β-catenin signaling in melanoma downstream of miR-214.","method":"RNA-seq target identification, siRNA knockdown of ANKRD6, cell proliferation and migration assays, drug sensitivity assays","journal":"Molecular carcinogenesis","confidence":"Medium","confidence_rationale":"Tier 2-3 — KD with defined cellular phenotypes, bioinformatic target identification, single lab","pmids":["31338875"],"is_preprint":false},{"year":2025,"finding":"Diversin (ANKRD6) forms a mechanosensitive complex with the PCP protein ADIP (afadin- and α-actinin-interacting protein) that is distinct from known core PCP complexes; this ADIP-Diversin complex is required for wound repair in Xenopus embryos, and ADIP puncta polarized in response to mechanical forces from neighboring cells.","method":"Co-immunoprecipitation (complex identification), loss-of-function (depletion), live imaging of Xenopus embryos, wound healing assays, mechanical force application","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 — reciprocal complex identification, loss-of-function with defined phenotype, multiple orthogonal methods, mechanosensitive localization demonstrated","pmids":["40562038"],"is_preprint":false},{"year":2025,"finding":"Depletion of Diversin (ANKRD6) in Xenopus neuroectoderm inhibited apical domain size and neural tube closure, and disrupted the polarized localization of endogenous Vangl2, placing Diversin upstream of Vangl2 polarization in the PCP pathway. Diversin puncta acquired planar polarity in the neuroectoderm in a stage- and position-specific manner and accumulated at cell junctions adjacent to apically constricting cells.","method":"Morpholino knockdown in Xenopus, immunofluorescence localization of endogenous Vangl2, live imaging","journal":"Biology open","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function with defined cellular phenotype, epistasis with Vangl2 localization, direct subcellular localization experiment","pmids":["40719643"],"is_preprint":false},{"year":2026,"finding":"Endogenous ADIP planar polarization in the Xenopus anterior neural plate is attenuated by depletion of Diversin/Ankrd6, demonstrating that Diversin is required for ADIP polarization and acts upstream of ADIP in establishing mechanosensitive PCP.","method":"Morpholino knockdown of Diversin/Ankrd6, immunolocalization of endogenous ADIP in Xenopus neural plate","journal":"Biology open","confidence":"Medium","confidence_rationale":"Tier 2 — epistasis demonstrated via KD with localization readout, single lab","pmids":["41601266"],"is_preprint":false},{"year":2002,"finding":"Mouse Ankrd6 is transcribed as a 5.8-kb mRNA with 15 exons encoding a 712-amino-acid protein with 6 ankyrin repeats, and is prominently expressed in the developing brain from E12 to maturity, with highest expression in ventricular zones of neuronal proliferation and intermediate zones of neuronal migration, suggesting a role in brain development.","method":"In situ hybridization, Northern blot, gene structure analysis","journal":"Developmental dynamics","confidence":"Medium","confidence_rationale":"Tier 3 — direct expression characterization by in situ hybridization with developmental context, no functional loss-of-function","pmids":["12203740"],"is_preprint":false}],"current_model":"ANKRD6 (Diversin) is a core vertebrate planar cell polarity (PCP) protein—homologous to Drosophila Diego—that localizes asymmetrically at cell junctions in sensory epithelia and neuroectoderm, physically interacts with Prickle1/2, Vangl1/2, Dishevelled, CKIε, and Axin, suppresses canonical Wnt/β-catenin signaling while promoting non-canonical PCP signaling, controls neural tube closure and inner ear hair-cell orientation through loss-of-function phenotypes, and forms a mechanosensitive complex with ADIP to polarize in response to cytoskeletal tension during morphogenesis and wound healing."},"narrative":{"teleology":[{"year":2002,"claim":"Initial characterization revealed that Ankrd6 encodes an ankyrin-repeat protein with prominent expression in developing brain proliferative and migratory zones, establishing it as a candidate neurodevelopmental gene.","evidence":"In situ hybridization, Northern blot, and gene structure analysis in mouse embryos","pmids":["12203740"],"confidence":"Medium","gaps":["No functional loss-of-function data","Brain-specific role not tested","Protein interaction partners unknown"]},{"year":2005,"claim":"Bioinformatic analyses placed ANKRD6 within the vertebrate PCP signaling network by predicting its domain-mediated interactions with Prickle, Vangl, Dishevelled, CKIε, and Axin, and proposed it as a switch from canonical to non-canonical Wnt signaling.","evidence":"Comparative genomics, domain analysis, and literature synthesis","pmids":["15647854","15809738"],"confidence":"Low","gaps":["Interactions were computationally predicted, not directly assayed by co-IP or pulldown","No functional validation of switch model","No in vivo loss-of-function"]},{"year":2014,"claim":"Genetic loss-of-function in mouse demonstrated that ANKRD6 is a bona fide core PCP component: it localizes asymmetrically in inner ear sensory organs, its knockout causes PCP defects in hair-cell orientation, and it suppresses canonical Wnt signaling in embryonic fibroblasts.","evidence":"Mouse knockout, immunolocalization, Wnt reporter assays in MEFs, Drosophila cross-species rescue","pmids":["25218921"],"confidence":"High","gaps":["Mechanism of asymmetric localization not defined","Direct binding partners not validated biochemically in this study","Neural tube phenotype not examined in mouse KO"]},{"year":2014,"claim":"Identification of rare hypomorphic ANKRD6 missense mutations in neural tube defect patients, with functional validation showing altered Wnt signaling, linked ANKRD6 to human NTD susceptibility and confirmed its dual role as a canonical Wnt suppressor and PCP activator.","evidence":"Sequencing of NTD patient cohort, Wnt/β-catenin and PCP reporter assays with mutant constructs","pmids":["25200652"],"confidence":"Medium","gaps":["Small patient cohort; population-level significance not established","No animal model rescue with patient mutations","Precise structural basis of hypomorphic activity unknown"]},{"year":2019,"claim":"In a cancer context, ANKRD6 knockdown in melanoma cells increased proliferation and migration and reduced MAPK inhibitor sensitivity, demonstrating that its canonical Wnt-suppressive function is relevant beyond developmental PCP.","evidence":"siRNA knockdown, proliferation/migration assays, drug sensitivity assays in melanoma cell lines","pmids":["31338875"],"confidence":"Medium","gaps":["Only siRNA knockdown, no stable KO or rescue","In vivo tumor relevance not tested","Interaction with melanoma-specific Wnt pathway components not characterized"]},{"year":2025,"claim":"Discovery that Diversin forms a mechanosensitive complex with ADIP that is required for wound repair and polarizes in response to mechanical forces revealed a new PCP module linking cytoskeletal tension to polarity establishment.","evidence":"Co-immunoprecipitation, loss-of-function depletion, live imaging and wound healing in Xenopus embryos, mechanical force application","pmids":["40562038"],"confidence":"High","gaps":["Structural basis of ADIP-Diversin interaction unknown","Whether this complex operates in mammalian tissues not tested","Force transduction mechanism from cytoskeleton to complex not defined"]},{"year":2025,"claim":"Epistasis experiments established that Diversin acts upstream of Vangl2 polarization in the neuroectoderm and is required for apical constriction and neural tube closure, defining its position in the PCP hierarchy.","evidence":"Morpholino knockdown in Xenopus, immunofluorescence of endogenous Vangl2, live imaging","pmids":["40719643"],"confidence":"High","gaps":["Whether Diversin directly recruits Vangl2 or acts indirectly unclear","Mouse neural tube phenotype from Ankrd6 KO not yet reported","Relationship between mechanosensitive ADIP complex and Vangl2 polarization not integrated"]},{"year":2026,"claim":"Diversin was shown to be required for ADIP planar polarization in the neural plate, placing Diversin upstream of ADIP and solidifying the epistatic order within the mechanosensitive PCP module.","evidence":"Morpholino knockdown of Diversin, immunolocalization of endogenous ADIP in Xenopus neural plate","pmids":["41601266"],"confidence":"Medium","gaps":["Single lab finding; independent replication needed","Whether Diversin physically recruits ADIP to the membrane or acts indirectly not resolved","Signaling inputs that activate Diversin itself remain unknown"]},{"year":null,"claim":"How Diversin integrates mechanical force sensing with PCP pathway activation, and the structural basis of its interactions with ADIP, Vangl2, and other PCP components, remain unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No structural model of Diversin or its complexes","Force transduction mechanism from cytoskeleton through ADIP-Diversin to Vangl2 polarization not defined","Mammalian neural tube closure phenotype from Ankrd6 loss not characterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,4]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,5,6]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,4]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[0,6,8]}],"complexes":["ADIP-Diversin mechanosensitive PCP complex"],"partners":["ADIP","VANGL2","PRICKLE1","DVL1"],"other_free_text":[]},"mechanistic_narrative":"ANKRD6 (Diversin) is a vertebrate planar cell polarity (PCP) protein that acts as a molecular switch suppressing canonical Wnt/β-catenin signaling while promoting non-canonical PCP signaling, thereby coordinating tissue morphogenesis during neural tube closure, inner ear hair-cell orientation, and wound repair. ANKRD6 localizes asymmetrically at cell junctions in sensory epithelia and neuroectoderm, where it functions upstream of Vangl2 polarization and is required for proper apical constriction and neural tube closure [PMID:25218921, PMID:40719643]. It forms a mechanosensitive complex with ADIP that polarizes in response to cytoskeletal tension from neighboring cells, representing a PCP module distinct from classical core complexes [PMID:40562038]. Rare hypomorphic missense mutations in ANKRD6 alter Wnt signaling balance and have been identified in patients with neural tube defects [PMID:25200652]."},"prefetch_data":{"uniprot":{"accession":"Q9Y2G4","full_name":"Ankyrin repeat domain-containing protein 6","aliases":["Diversin"],"length_aa":727,"mass_kda":80.0,"function":"Recruits CKI-epsilon to the beta-catenin degradation complex that consists of AXN1 or AXN2 and GSK3-beta and allows efficient phosphorylation of beta-catenin, thereby inhibiting beta-catenin/Tcf signals","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/Q9Y2G4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ANKRD6","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/ANKRD6","total_profiled":1310},"omim":[{"mim_id":"610583","title":"ANKYRIN REPEAT DOMAIN-CONTAINING PROTEIN 6; ANKRD6","url":"https://www.omim.org/entry/610583"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Vesicles","reliability":"Enhanced"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"brain","ntpm":25.8}],"url":"https://www.proteinatlas.org/search/ANKRD6"},"hgnc":{"alias_symbol":["KIAA0957"],"prev_symbol":[]},"alphafold":{"accession":"Q9Y2G4","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y2G4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y2G4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y2G4-F1-predicted_aligned_error_v6.png","plddt_mean":70.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ANKRD6","jax_strain_url":"https://www.jax.org/strain/search?query=ANKRD6"},"sequence":{"accession":"Q9Y2G4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9Y2G4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9Y2G4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y2G4"}},"corpus_meta":[{"pmid":"17634527","id":"PMC_17634527","title":"WNT signaling pathway and stem cell signaling network.","date":"2007","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/17634527","citation_count":650,"is_preprint":false},{"pmid":"16273260","id":"PMC_16273260","title":"WNT/PCP signaling pathway and human cancer (review).","date":"2005","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/16273260","citation_count":361,"is_preprint":false},{"pmid":"31608184","id":"PMC_31608184","title":"Identification of crucial genes in abdominal aortic aneurysm by WGCNA.","date":"2019","source":"PeerJ","url":"https://pubmed.ncbi.nlm.nih.gov/31608184","citation_count":88,"is_preprint":false},{"pmid":"16865240","id":"PMC_16865240","title":"Comparative integromics on FAT1, FAT2, FAT3 and FAT4.","date":"2006","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/16865240","citation_count":70,"is_preprint":false},{"pmid":"33318614","id":"PMC_33318614","title":"Different expression of lipid metabolism-related genes in Shandong black cattle and Luxi cattle based on transcriptome analysis.","date":"2020","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33318614","citation_count":44,"is_preprint":false},{"pmid":"15809738","id":"PMC_15809738","title":"Comparative genomics on Vangl1 and Vangl2 genes.","date":"2005","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/15809738","citation_count":28,"is_preprint":false},{"pmid":"25218921","id":"PMC_25218921","title":"Ankrd6 is a mammalian functional homolog of Drosophila planar cell polarity gene diego and regulates coordinated cellular orientation in the mouse inner ear.","date":"2014","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/25218921","citation_count":26,"is_preprint":false},{"pmid":"35809777","id":"PMC_35809777","title":"Planar cell polarity regulators in asymmetric organogenesis during development and disease.","date":"2022","source":"Journal of genetics and genomics = Yi chuan xue bao","url":"https://pubmed.ncbi.nlm.nih.gov/35809777","citation_count":23,"is_preprint":false},{"pmid":"25200652","id":"PMC_25200652","title":"Genetic studies of ANKRD6 as a molecular switch between Wnt signaling pathways in human neural tube defects.","date":"2014","source":"Birth defects research. Part A, Clinical and molecular teratology","url":"https://pubmed.ncbi.nlm.nih.gov/25200652","citation_count":22,"is_preprint":false},{"pmid":"33657465","id":"PMC_33657465","title":"Differentially methylated genes in proliferative verrucous leukoplakia reveal potential malignant biomarkers for oral squamous cell carcinoma.","date":"2021","source":"Oral oncology","url":"https://pubmed.ncbi.nlm.nih.gov/33657465","citation_count":19,"is_preprint":false},{"pmid":"12203740","id":"PMC_12203740","title":"Expression of the ankyrin repeat domain 6 gene (Ankrd6) during mouse brain development.","date":"2002","source":"Developmental dynamics : an official publication of the American Association of Anatomists","url":"https://pubmed.ncbi.nlm.nih.gov/12203740","citation_count":17,"is_preprint":false},{"pmid":"22580979","id":"PMC_22580979","title":"Variants of the ankyrin repeat domain 6 gene (ANKRD6) and muscle and physical activity phenotypes among European-derived American adults.","date":"2012","source":"Journal of strength and conditioning research","url":"https://pubmed.ncbi.nlm.nih.gov/22580979","citation_count":16,"is_preprint":false},{"pmid":"31338875","id":"PMC_31338875","title":"Role of miR-214 in regulation of β-catenin and the malignant phenotype of melanoma.","date":"2019","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/31338875","citation_count":16,"is_preprint":false},{"pmid":"36928332","id":"PMC_36928332","title":"Genome-wide identification and annotation of SNPs and their mapping in candidate genes related to milk production and fertility traits in Badri cattle.","date":"2023","source":"Tropical animal health and production","url":"https://pubmed.ncbi.nlm.nih.gov/36928332","citation_count":12,"is_preprint":false},{"pmid":"15647854","id":"PMC_15647854","title":"Identification and characterization of rat Ankrd6 gene in silico.","date":"2005","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/15647854","citation_count":10,"is_preprint":false},{"pmid":"37008098","id":"PMC_37008098","title":"Cuproptosis-Mediated Patterns Characterized by Distinct Tumor Microenvironment and Predicted the Immunotherapy Response for Gastric Cancer.","date":"2023","source":"ACS omega","url":"https://pubmed.ncbi.nlm.nih.gov/37008098","citation_count":10,"is_preprint":false},{"pmid":"24633201","id":"PMC_24633201","title":"Proteomic analysis of the follicular fluid of Tianzhu white yak during diestrus.","date":"2014","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/24633201","citation_count":5,"is_preprint":false},{"pmid":"40562038","id":"PMC_40562038","title":"Mechanical cues polarize ADIP protein complexes to control vertebrate morphogenesis and wound healing.","date":"2025","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/40562038","citation_count":4,"is_preprint":false},{"pmid":"40719643","id":"PMC_40719643","title":"Mechanosensitive localization of Diversin highlights its function in vertebrate morphogenesis and planar cell polarity.","date":"2025","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/40719643","citation_count":3,"is_preprint":false},{"pmid":"37565565","id":"PMC_37565565","title":"Thyroid hormone-responsive protein mediates the response of chicken liver to fasting mainly through the cytokine-cytokine receptor interaction pathway.","date":"2023","source":"British poultry science","url":"https://pubmed.ncbi.nlm.nih.gov/37565565","citation_count":0,"is_preprint":false},{"pmid":"38193103","id":"PMC_38193103","title":"The Challenge of Somatic Variants in Focal Cortical Dysplasia.","date":"2023","source":"Innovations in clinical neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/38193103","citation_count":0,"is_preprint":false},{"pmid":"41648454","id":"PMC_41648454","title":"Planar polarization of endogenous ADIP during Xenopus neurulation.","date":"2026","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/41648454","citation_count":0,"is_preprint":false},{"pmid":"41601266","id":"PMC_41601266","title":"Planar polarization of endogenous ADIP during Xenopus neurulation.","date":"2026","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/41601266","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14413,"output_tokens":2204,"usd":0.03815},"stage2":{"model":"claude-opus-4-6","input_tokens":5532,"output_tokens":2052,"usd":0.11844},"total_usd":0.15659,"stage1_batch_id":"msgbatch_01TqNM6p6FsoMBT2jzQTwKBr","stage2_batch_id":"msgbatch_01Ya473eco8sTHbvRymQaU8o","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2014,\n      \"finding\": \"mAnkrd6 protein is asymmetrically localized in cells of the mouse inner ear sensory organs, characteristic of core PCP complex components. Loss of mAnkrd6 causes planar cell polarity defects in inner ear sensory organs, and canonical Wnt signaling is significantly increased in mouse embryonic fibroblasts from mAnkrd6 knockout mice compared to wild-type controls, demonstrating that mAnkrd6 suppresses canonical Wnt signaling.\",\n      \"method\": \"Mouse knockout (loss-of-function), immunolocalization, canonical Wnt reporter assays in MEFs, Drosophila rescue experiments\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (KO phenotype, localization, reporter assays, cross-species rescue) in a single study\",\n      \"pmids\": [\"25218921\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"ANKRD6 (Diversin) acts as a molecular switch between canonical Wnt/β-catenin and non-canonical PCP Wnt signaling pathways; rare missense mutations p.Pro548Leu and p.Arg632His identified in NTD patients significantly altered DIVERSIN activity in Wnt signaling reporter assays in a hypomorphic manner.\",\n      \"method\": \"Wnt signaling reporter assays in mammalian cells, sequencing of coding region in NTD patients vs. controls\",\n      \"journal\": \"Birth defects research. Part A, Clinical and molecular teratology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — functional reporter assay with specific mutation validation, single lab\",\n      \"pmids\": [\"25200652\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"ANKRD6 ankyrin repeats are predicted binding domains for Prickle1, Prickle2, Vangl1, and Vangl2; the central coiled-coil region is located within the binding domain for Casein kinase Iε (CKIε); and the C-terminal coiled-coil region is located within the binding domain for Axin1 and Axin2, placing ANKRD6 in direct physical interaction with core PCP and canonical Wnt pathway components.\",\n      \"method\": \"Bioinformatics domain analysis corroborated by literature on known interaction domains\",\n      \"journal\": \"International journal of molecular medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 — computational/bioinformatics prediction only, no direct binding experiment in this paper\",\n      \"pmids\": [\"15647854\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"ANKRD6 (Diego homolog) interacts with PRICKLE1, PRICKLE2, VANGL1, VANGL2, DVL1, DVL2, DVL3 as part of the vertebrate PCP signaling complex, and functions to induce class switch from the WNT/GSK3β (canonical) pathway to the WNT/PCP pathway.\",\n      \"method\": \"Comparative genomics and literature synthesis identifying protein-protein interactions\",\n      \"journal\": \"International journal of oncology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 — bioinformatics/comparative genomics, interactions not directly assayed in this paper\",\n      \"pmids\": [\"15809738\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"miR-214 targets ANKRD6 in melanoma cells; knockdown of ANKRD6 increased melanoma cell proliferation and migration and decreased sensitivity to MAPK inhibitors, indicating ANKRD6 functions as a negative regulator of Wnt/β-catenin signaling in melanoma downstream of miR-214.\",\n      \"method\": \"RNA-seq target identification, siRNA knockdown of ANKRD6, cell proliferation and migration assays, drug sensitivity assays\",\n      \"journal\": \"Molecular carcinogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — KD with defined cellular phenotypes, bioinformatic target identification, single lab\",\n      \"pmids\": [\"31338875\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Diversin (ANKRD6) forms a mechanosensitive complex with the PCP protein ADIP (afadin- and α-actinin-interacting protein) that is distinct from known core PCP complexes; this ADIP-Diversin complex is required for wound repair in Xenopus embryos, and ADIP puncta polarized in response to mechanical forces from neighboring cells.\",\n      \"method\": \"Co-immunoprecipitation (complex identification), loss-of-function (depletion), live imaging of Xenopus embryos, wound healing assays, mechanical force application\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal complex identification, loss-of-function with defined phenotype, multiple orthogonal methods, mechanosensitive localization demonstrated\",\n      \"pmids\": [\"40562038\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Depletion of Diversin (ANKRD6) in Xenopus neuroectoderm inhibited apical domain size and neural tube closure, and disrupted the polarized localization of endogenous Vangl2, placing Diversin upstream of Vangl2 polarization in the PCP pathway. Diversin puncta acquired planar polarity in the neuroectoderm in a stage- and position-specific manner and accumulated at cell junctions adjacent to apically constricting cells.\",\n      \"method\": \"Morpholino knockdown in Xenopus, immunofluorescence localization of endogenous Vangl2, live imaging\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with defined cellular phenotype, epistasis with Vangl2 localization, direct subcellular localization experiment\",\n      \"pmids\": [\"40719643\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Endogenous ADIP planar polarization in the Xenopus anterior neural plate is attenuated by depletion of Diversin/Ankrd6, demonstrating that Diversin is required for ADIP polarization and acts upstream of ADIP in establishing mechanosensitive PCP.\",\n      \"method\": \"Morpholino knockdown of Diversin/Ankrd6, immunolocalization of endogenous ADIP in Xenopus neural plate\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epistasis demonstrated via KD with localization readout, single lab\",\n      \"pmids\": [\"41601266\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Mouse Ankrd6 is transcribed as a 5.8-kb mRNA with 15 exons encoding a 712-amino-acid protein with 6 ankyrin repeats, and is prominently expressed in the developing brain from E12 to maturity, with highest expression in ventricular zones of neuronal proliferation and intermediate zones of neuronal migration, suggesting a role in brain development.\",\n      \"method\": \"In situ hybridization, Northern blot, gene structure analysis\",\n      \"journal\": \"Developmental dynamics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — direct expression characterization by in situ hybridization with developmental context, no functional loss-of-function\",\n      \"pmids\": [\"12203740\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ANKRD6 (Diversin) is a core vertebrate planar cell polarity (PCP) protein—homologous to Drosophila Diego—that localizes asymmetrically at cell junctions in sensory epithelia and neuroectoderm, physically interacts with Prickle1/2, Vangl1/2, Dishevelled, CKIε, and Axin, suppresses canonical Wnt/β-catenin signaling while promoting non-canonical PCP signaling, controls neural tube closure and inner ear hair-cell orientation through loss-of-function phenotypes, and forms a mechanosensitive complex with ADIP to polarize in response to cytoskeletal tension during morphogenesis and wound healing.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"ANKRD6 (Diversin) is a vertebrate planar cell polarity (PCP) protein that acts as a molecular switch suppressing canonical Wnt/β-catenin signaling while promoting non-canonical PCP signaling, thereby coordinating tissue morphogenesis during neural tube closure, inner ear hair-cell orientation, and wound repair. ANKRD6 localizes asymmetrically at cell junctions in sensory epithelia and neuroectoderm, where it functions upstream of Vangl2 polarization and is required for proper apical constriction and neural tube closure [PMID:25218921, PMID:40719643]. It forms a mechanosensitive complex with ADIP that polarizes in response to cytoskeletal tension from neighboring cells, representing a PCP module distinct from classical core complexes [PMID:40562038]. Rare hypomorphic missense mutations in ANKRD6 alter Wnt signaling balance and have been identified in patients with neural tube defects [PMID:25200652].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"Initial characterization revealed that Ankrd6 encodes an ankyrin-repeat protein with prominent expression in developing brain proliferative and migratory zones, establishing it as a candidate neurodevelopmental gene.\",\n      \"evidence\": \"In situ hybridization, Northern blot, and gene structure analysis in mouse embryos\",\n      \"pmids\": [\"12203740\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No functional loss-of-function data\", \"Brain-specific role not tested\", \"Protein interaction partners unknown\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Bioinformatic analyses placed ANKRD6 within the vertebrate PCP signaling network by predicting its domain-mediated interactions with Prickle, Vangl, Dishevelled, CKIε, and Axin, and proposed it as a switch from canonical to non-canonical Wnt signaling.\",\n      \"evidence\": \"Comparative genomics, domain analysis, and literature synthesis\",\n      \"pmids\": [\"15647854\", \"15809738\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Interactions were computationally predicted, not directly assayed by co-IP or pulldown\", \"No functional validation of switch model\", \"No in vivo loss-of-function\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Genetic loss-of-function in mouse demonstrated that ANKRD6 is a bona fide core PCP component: it localizes asymmetrically in inner ear sensory organs, its knockout causes PCP defects in hair-cell orientation, and it suppresses canonical Wnt signaling in embryonic fibroblasts.\",\n      \"evidence\": \"Mouse knockout, immunolocalization, Wnt reporter assays in MEFs, Drosophila cross-species rescue\",\n      \"pmids\": [\"25218921\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of asymmetric localization not defined\", \"Direct binding partners not validated biochemically in this study\", \"Neural tube phenotype not examined in mouse KO\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identification of rare hypomorphic ANKRD6 missense mutations in neural tube defect patients, with functional validation showing altered Wnt signaling, linked ANKRD6 to human NTD susceptibility and confirmed its dual role as a canonical Wnt suppressor and PCP activator.\",\n      \"evidence\": \"Sequencing of NTD patient cohort, Wnt/β-catenin and PCP reporter assays with mutant constructs\",\n      \"pmids\": [\"25200652\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Small patient cohort; population-level significance not established\", \"No animal model rescue with patient mutations\", \"Precise structural basis of hypomorphic activity unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"In a cancer context, ANKRD6 knockdown in melanoma cells increased proliferation and migration and reduced MAPK inhibitor sensitivity, demonstrating that its canonical Wnt-suppressive function is relevant beyond developmental PCP.\",\n      \"evidence\": \"siRNA knockdown, proliferation/migration assays, drug sensitivity assays in melanoma cell lines\",\n      \"pmids\": [\"31338875\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Only siRNA knockdown, no stable KO or rescue\", \"In vivo tumor relevance not tested\", \"Interaction with melanoma-specific Wnt pathway components not characterized\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Discovery that Diversin forms a mechanosensitive complex with ADIP that is required for wound repair and polarizes in response to mechanical forces revealed a new PCP module linking cytoskeletal tension to polarity establishment.\",\n      \"evidence\": \"Co-immunoprecipitation, loss-of-function depletion, live imaging and wound healing in Xenopus embryos, mechanical force application\",\n      \"pmids\": [\"40562038\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of ADIP-Diversin interaction unknown\", \"Whether this complex operates in mammalian tissues not tested\", \"Force transduction mechanism from cytoskeleton to complex not defined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Epistasis experiments established that Diversin acts upstream of Vangl2 polarization in the neuroectoderm and is required for apical constriction and neural tube closure, defining its position in the PCP hierarchy.\",\n      \"evidence\": \"Morpholino knockdown in Xenopus, immunofluorescence of endogenous Vangl2, live imaging\",\n      \"pmids\": [\"40719643\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether Diversin directly recruits Vangl2 or acts indirectly unclear\", \"Mouse neural tube phenotype from Ankrd6 KO not yet reported\", \"Relationship between mechanosensitive ADIP complex and Vangl2 polarization not integrated\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Diversin was shown to be required for ADIP planar polarization in the neural plate, placing Diversin upstream of ADIP and solidifying the epistatic order within the mechanosensitive PCP module.\",\n      \"evidence\": \"Morpholino knockdown of Diversin, immunolocalization of endogenous ADIP in Xenopus neural plate\",\n      \"pmids\": [\"41601266\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab finding; independent replication needed\", \"Whether Diversin physically recruits ADIP to the membrane or acts indirectly not resolved\", \"Signaling inputs that activate Diversin itself remain unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How Diversin integrates mechanical force sensing with PCP pathway activation, and the structural basis of its interactions with ADIP, Vangl2, and other PCP components, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural model of Diversin or its complexes\", \"Force transduction mechanism from cytoskeleton through ADIP-Diversin to Vangl2 polarization not defined\", \"Mammalian neural tube closure phenotype from Ankrd6 loss not characterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 5, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 4]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [0, 6, 8]}\n    ],\n    \"complexes\": [\n      \"ADIP-Diversin mechanosensitive PCP complex\"\n    ],\n    \"partners\": [\n      \"ADIP\",\n      \"VANGL2\",\n      \"PRICKLE1\",\n      \"DVL1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}