{"gene":"ANXA6","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":2020,"finding":"ANXA6 promotes autophagy by inhibiting the PI3K/AKT/mTOR signaling pathway in nasopharyngeal carcinoma cells; siRNA knockdown of ANXA6 activated PI3K/AKT/mTOR and suppressed autophagy, and co-treatment with PI3K/AKT/mTOR inhibitor CAL101 reversed siANXA6-reduced autophagy, establishing negative regulatory feedback.","method":"siRNA knockdown, pharmacological inhibition (CAL101), western blotting of pathway components","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined molecular phenotype and pharmacological rescue, single lab, two orthogonal perturbation methods","pmids":["32373608"],"is_preprint":false},{"year":2020,"finding":"ANXA6 enables ATG9A-positive vesicle trafficking from recycling endosomes to autophagosomes through RAB GTPases and F-actin, thereby facilitating autophagosome formation; ANXA6 also suppresses mTOR activity by inhibiting PI3K-AKT and ERK signaling pathways.","method":"siRNA knockdown, re-expression rescue, GO analysis of interactors, confocal imaging of ATG9A vesicle trafficking, western blotting of mTOR pathway","journal":"Clinical and translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function plus rescue with defined trafficking phenotype, single lab, multiple orthogonal readouts","pmids":["33135350"],"is_preprint":false},{"year":2023,"finding":"ANXA6 is SUMOylated by SUMO1 at lysine K299; this SUMOylation promotes ANXA6 binding to EGFR upon EGF stimulation and facilitates EGFR-PKCα complex formation, which decreases phosphorylation of EGFR and ERK1/2. The K299R mutant (SUMO-deficient) loses the ability to suppress EGFR-ERK signaling and confers drug resistance to gefitinib.","method":"Co-transfection with SUMO1/UBC9/His-SUMO1 plasmids, immunoprecipitation, LC-MS/MS identification of SUMOylation sites, site-directed mutagenesis (K299R), shRNA knockdown, lentiviral overexpression, xenograft mouse model","journal":"Cell communication and signaling : CCS","confidence":"High","confidence_rationale":"Tier 1 / Strong — SUMOylation site identified by MS and validated by mutagenesis, complex formation confirmed by Co-IP, functional consequence tested in vitro and in vivo","pmids":["37528485"],"is_preprint":false},{"year":2020,"finding":"The EHEC effector protein EspF interacts with host protein ANXA6 through EspF's C-terminal domain; this EspF-ANXA6 interaction activates MLCK, induces MLC and PKCα phosphorylation, downregulates calmodulin, and disrupts tight junction proteins ZO-1 and occludin, leading to intestinal barrier disruption.","method":"FRET, co-immunoprecipitation, domain mapping, western blotting, immunofluorescence of tight junction distribution","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — interaction confirmed by two orthogonal methods (FRET + Co-IP), downstream pathway assessed by western blot, single lab","pmids":["33425920"],"is_preprint":false},{"year":2018,"finding":"AnxA6-knockout mice on a high-fat diet show increased insulin-stimulated Akt and S6K phosphorylation in liver but fail to suppress hepatic gluconeogenesis upon insulin exposure; AnxA6-depleted HuH7 hepatocytes also fail to reduce glucose production upon insulin, implicating AnxA6 in fine-tuning hepatic glucose metabolism downstream of or parallel to insulin signaling.","method":"AnxA6-KO mouse model, OGTT, ITT, western blotting of phospho-Akt and phospho-S6, glucose production assay in AnxA6-depleted hepatocytes","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mouse phenotype corroborated by in vitro hepatocyte loss-of-function, multiple metabolic readouts, single lab","pmids":["30110341"],"is_preprint":false},{"year":2025,"finding":"Anxa6-knockout mice display rapid fasting hypoglycaemia due to inability to utilize alanine for hepatic gluconeogenesis (glucose-alanine cycle impairment), associated with reduced expression of hepatic alanine aminotransferase 2 (Gpt2), lactate dehydrogenase (Ldha2), and the alanine transporter SNAT4.","method":"Anxa6-/- mouse model, indirect calorimetry, glucose/insulin/alanine tolerance tests, biochemical analysis, gene expression profiling","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mouse with multiple metabolic tolerance tests and molecular pathway readouts, single lab, peer-reviewed","pmids":["40802799"],"is_preprint":false},{"year":2013,"finding":"In Niemann-Pick type C disease cells, AnxA6 translocates to cholesterol- and sphingomyelin-enriched lipid microdomains (DRMs) upon a rise in Ca2+ concentration; in vitro Langmuir monolayer experiments showed Ca2+ is the primary factor regulating AnxA6 interaction with DPPC/sphingomyelin monolayers and its localization in these microdomains.","method":"Lipid microdomain (DRM) isolation by fractionation, Langmuir monolayer assay with defined lipid compositions, Ca2+ titration","journal":"Molecular bioSystems","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — in vitro reconstitution with Langmuir monolayers plus cellular fractionation, single lab, two orthogonal methods","pmids":["23360953"],"is_preprint":false},{"year":2022,"finding":"EZH2 suppresses ANXA6 expression by increasing H3K27me3 histone methylation at the ANXA6 promoter; overexpression of EZH2 reduced ANXA6 levels, inhibited Ang II-induced ROS, and protected against vascular smooth muscle cell senescence, while co-overexpression of ANXA6 partially reversed the EZH2 protective effect.","method":"ChIP (H3K27me3 at ANXA6 promoter), lentiviral overexpression of EZH2 and ANXA6, SA-β-gal staining, flow cytometry, ROS assay, western blotting","journal":"Oxidative medicine and cellular longevity","confidence":"Low","confidence_rationale":"Tier 2 / Weak — ChIP validates epigenetic mechanism but paper was subsequently retracted (PMID:38188996); evidence is unreliable","pmids":["36160712","38188996"],"is_preprint":false},{"year":2021,"finding":"CRISPR-Cas9 knockout of ANXA6 in HEK293FT cells significantly increased influenza A virus replication and viral titer, demonstrating that AnxA6 acts as a host restriction factor counteracting influenza virion packaging.","method":"CRISPR-Cas9 gene knockout, TCID50 assay, flow cytometry, viral growth curve experiments","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO with defined functional phenotype (viral restriction), two quantitative methods, single lab","pmids":["34673207"],"is_preprint":false},{"year":2022,"finding":"Apigenin disrupts intracellular distribution of AnxA6 (blocking AnxA6 aggregation) and TNAP attachment to membranes in osteoblast cell lines, thereby modulating matrix vesicle-mediated mineralization; the effects were cell-type dependent across hFOB 1.19, Saos-2, and HCASMC cells.","method":"Fluorescence microscopy of AnxA6 distribution, TEM-gold immunolabeling of vesicles, AR-S/CPC mineralization assay, ELISA for TNAP activity","journal":"International journal of molecular sciences","confidence":"Low","confidence_rationale":"Tier 3 / Weak — localization experiments with functional correlation but no direct mechanistic perturbation of AnxA6, single lab","pmids":["36361965"],"is_preprint":false},{"year":2023,"finding":"ANXA6 interacts with TRPV2; ANXA6 induces autophagy in head and neck squamous cell carcinoma by inhibiting the AKT/mTOR signaling pathway, and TRPV2 inhibition reverses ANXA6-induced autophagy and lymphatic metastasis, placing TRPV2 downstream of ANXA6 in this pathway.","method":"ANXA6 overexpression/knockdown in vitro and in vivo (lymphatic metastasis model), Co-IP/molecular interaction assays, AKT/mTOR pathway western blotting, TRPV2 pharmacological inhibition","journal":"Experimental hematology & oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — interaction shown with Co-IP, epistasis demonstrated by pharmacological rescue, in vivo model, single lab","pmids":["37138336"],"is_preprint":false},{"year":2024,"finding":"PRRX1 transcription factor directly promotes ANXA6 transcription (validated by ChIP-PCR and dual-luciferase assay); ANXA6 in turn promotes PKCα/EGFR complex formation by co-immunoprecipitation, inhibiting EGFR phosphorylation and downstream AKT and ERK1/2, thereby enhancing cisplatin sensitivity in bladder cancer cells.","method":"ChIP-PCR, dual-luciferase reporter assay, mass spectrometry, immunofluorescence, co-immunoprecipitation, xenograft model","journal":"Life sciences","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — transcription factor binding validated by ChIP and luciferase; EGFR-PKCα complex by Co-IP and MS; single lab with multiple methods","pmids":["39528080"],"is_preprint":false},{"year":2025,"finding":"ANXA6 co-localizes with cGAS and promotes ubiquitination (K63-linked) of cGAS, suppressing the cGAS-STING signaling pathway; ANXA6 knockdown inhibits K63-linked ubiquitination of cGAS, leading to cGAS accumulation, activation of phospho-TBK1/IRF3, type I IFN-stimulated genes, increased ROS and lipid peroxides, and ultimately ferroptosis in cervical cancer cells.","method":"Co-localization (immunofluorescence), western blotting of cGAS-STING pathway components, ubiquitination assay, ROS and lipid peroxide measurements, ANXA6 knockdown","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-localization and ubiquitination assay establish mechanism, pathway validated by downstream signaling, single lab","pmids":["41219880"],"is_preprint":false},{"year":2025,"finding":"MG53 (mitsugumin 53) binds AnxA6 and exerts cytoprotective effects on iPSCs under hypoxia/reoxygenation injury; overexpression of AnxA6 abolishes MG53's membrane-stabilizing cytoprotective effects, indicating AnxA6 negatively modulates MG53-mediated membrane repair.","method":"Co-immunoprecipitation (MG53-AnxA6 binding), AnxA6 overexpression rescue experiment, cell viability assays, in vivo iPSC transplantation in MIRI model","journal":"Cellular signalling","confidence":"Low","confidence_rationale":"Tier 3 / Weak — binding shown by Co-IP but mechanistic detail is limited; single lab, limited orthogonal validation","pmids":["41106752"],"is_preprint":false},{"year":2025,"finding":"ANXA6 overexpression in decidual macrophages promotes mitochondrial ROS accumulation and reduces mitochondrial membrane potential via the NF-κB signaling pathway, triggering NLRP3-dependent pyroptosis and M1 macrophage polarization; conditioned medium from ANXA6-overexpressing macrophages inhibits trophoblast function via TNF-α-mediated suppression of the PI3K/AKT axis.","method":"ANXA6 overexpression in primary macrophages, ROS assay, mitochondrial membrane potential assay, NLRP3 inhibitor rescue, NF-κB pathway analysis, in vitro trophoblast co-culture","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss/gain of function with defined molecular pathway, pharmacological rescue with NLRP3 inhibitor and ROS scavenger, single lab","pmids":["40959268"],"is_preprint":false},{"year":2024,"finding":"AnxA6 interacts with SNAP23 and this interaction is required for the translocation of AnxA6 to cellular membranes, which facilitates secretion of small extracellular vesicles (EVs) and pro-inflammatory cytokines (MCP-1/CCL2, IL-8) in triple-negative breast cancer cells; AnxA6 downregulation strongly reduces EV secretion and associated cholesterol and MCP-1 content.","method":"Co-immunoprecipitation (AnxA6-SNAP23), cytokine arrays of secreted factors, EV isolation and characterization, AnxA6 knockdown and overexpression, cholesterol quantification in EVs","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 / Weak — Co-IP interaction plus functional secretion data, preprint not yet peer-reviewed, single lab","pmids":[],"is_preprint":true},{"year":2025,"finding":"In healthy skeletal muscle cells, mechanical stress induces upregulation of ANXA6 (and ANXA5), suggesting ANXA6 participates in an adaptive membrane repair response; in Duchenne muscular dystrophy (DMD) cells, ANXA6 expression is dysregulated, contributing to defective membrane repair.","method":"Live-cell imaging of GFP-tagged annexins, shear stress and laser irradiation injury assays, western blotting, immunohistochemistry of DMD patient biopsies","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 / Weak — localization and upregulation data from injury assays establish correlation with membrane repair; preprint, mechanistic detail for ANXA6 specifically is limited","pmids":[],"is_preprint":true}],"current_model":"ANXA6 (Annexin A6) is a Ca2+-dependent membrane- and phospholipid-binding scaffold protein that: (1) suppresses EGFR-ERK/AKT signaling by facilitating EGFR-PKCα complex formation (a function potentiated by SUMO1 modification at K299); (2) promotes autophagy by inhibiting PI3K/AKT/mTOR signaling and facilitating ATG9A vesicle trafficking via RAB GTPases and F-actin; (3) regulates cholesterol homeostasis by Ca2+-dependent translocation into lipid microdomains; (4) acts as a host restriction factor for influenza replication; (5) suppresses cGAS-STING signaling by promoting K63-linked ubiquitination of cGAS; (6) is required for hepatic alanine-dependent gluconeogenesis; and (7) facilitates secretion of extracellular vesicles and pro-inflammatory cytokines through interaction with SNAP23."},"narrative":{"mechanistic_narrative":"ANXA6 is a Ca2+-responsive membrane- and lipid-associated scaffold protein that integrates growth-factor signaling, autophagy, and membrane organization across diverse cell types [PMID:33135350, PMID:23360953]. Its best-characterized signaling output is suppression of the EGFR axis: ANXA6 promotes formation of an EGFR-PKCα complex that lowers EGFR and ERK1/2 phosphorylation, a function potentiated by SUMO1 conjugation at lysine K299, with the SUMO-deficient K299R mutant losing this activity and conferring gefitinib resistance [PMID:37528485]. The same EGFR-PKCα-restraining activity is engaged downstream of the transcription factor PRRX1 to enhance cisplatin sensitivity in bladder cancer [PMID:39528080]. ANXA6 also drives autophagy by inhibiting PI3K/AKT/mTOR (and ERK) signaling and by enabling ATG9A-vesicle trafficking from recycling endosomes to autophagosomes via RAB GTPases and F-actin [PMID:32373608, PMID:33135350], with TRPV2 acting downstream of ANXA6-induced autophagy [PMID:37138336]. Consistent with Ca2+-dependent lipid binding, ANXA6 translocates into cholesterol- and sphingomyelin-rich microdomains as intracellular Ca2+ rises [PMID:23360953], and supports SNAP23-dependent membrane translocation that facilitates secretion of extracellular vesicles and pro-inflammatory cytokines. Beyond these roles, ANXA6 acts as a host restriction factor limiting influenza A virus replication [PMID:34673207], suppresses cGAS-STING signaling by promoting K63-linked ubiquitination of cGAS [PMID:41219880], and is required in liver for insulin-modulated suppression of hepatic gluconeogenesis and for alanine-dependent gluconeogenesis [PMID:30110341, PMID:40802799].","teleology":[{"year":2013,"claim":"Established the biophysical basis for ANXA6 membrane targeting by showing that Ca2+ drives its translocation into cholesterol/sphingomyelin-enriched microdomains.","evidence":"DRM fractionation in Niemann-Pick type C cells plus Langmuir monolayer reconstitution with Ca2+ titration","pmids":["23360953"],"confidence":"Medium","gaps":["Does not identify the protein partners ANXA6 recruits to these microdomains","Functional consequence for cholesterol handling in vivo not resolved"]},{"year":2018,"claim":"Connected ANXA6 to systemic metabolism by showing knockout livers fail to suppress gluconeogenesis upon insulin, placing ANXA6 in the hepatic insulin response.","evidence":"AnxA6-KO mice with OGTT/ITT and phospho-Akt/S6 blotting, plus glucose-production assays in depleted hepatocytes","pmids":["30110341"],"confidence":"Medium","gaps":["Molecular target through which ANXA6 modulates Akt/S6K not defined","Direct binding partners in hepatocytes not identified"]},{"year":2020,"claim":"Defined ANXA6 as a positive regulator of autophagy operating both by inhibiting PI3K/AKT/mTOR and by physically enabling ATG9A vesicle delivery to autophagosomes.","evidence":"siRNA knockdown with re-expression rescue, pharmacological PI3K inhibition, and confocal imaging of ATG9A trafficking","pmids":["32373608","33135350"],"confidence":"Medium","gaps":["Whether ANXA6 binds ATG9A directly or acts through RAB/F-actin scaffolds is unresolved","How the mTOR-inhibitory and trafficking functions are coordinated is unclear"]},{"year":2020,"claim":"Identified ANXA6 as a host target of the EHEC effector EspF, linking it to tight-junction integrity via MLCK/PKCα signaling.","evidence":"FRET and Co-IP with domain mapping, plus tight-junction protein immunofluorescence","pmids":["33425920"],"confidence":"Medium","gaps":["Does not establish ANXA6's endogenous role at tight junctions absent infection","Direct enzymatic consequence of EspF binding to ANXA6 unknown"]},{"year":2021,"claim":"Showed ANXA6 functions as a host restriction factor against influenza A virus.","evidence":"CRISPR-Cas9 knockout in HEK293FT with TCID50 and viral growth-curve readouts","pmids":["34673207"],"confidence":"Medium","gaps":["Step of the viral life cycle ANXA6 acts on is not mechanistically pinned","No viral or host binding partner identified"]},{"year":2023,"claim":"Revealed that SUMO1 modification at K299 is required for ANXA6 to suppress EGFR-ERK signaling, mechanistically tying a post-translational modification to drug response.","evidence":"MS-mapped SUMOylation site validated by K299R mutagenesis, Co-IP of EGFR-PKCα complex, and xenograft model","pmids":["37528485"],"confidence":"High","gaps":["The SUMO E3 ligase and conditions controlling K299 modification are not defined","Structural basis of SUMO-dependent EGFR binding unknown"]},{"year":2023,"claim":"Extended the autophagy axis by identifying TRPV2 as a downstream effector of ANXA6-induced, AKT/mTOR-dependent autophagy in head and neck cancer metastasis.","evidence":"Gain/loss-of-function in vitro and in a lymphatic metastasis model, Co-IP, and TRPV2 pharmacological rescue","pmids":["37138336"],"confidence":"Medium","gaps":["Whether ANXA6-TRPV2 binding is direct and Ca2+-dependent is unresolved","Reciprocal validation of the interaction is limited"]},{"year":2024,"claim":"Placed the EGFR-suppressive ANXA6 function within a transcriptional circuit, showing PRRX1 directly drives ANXA6 expression to enhance chemosensitivity.","evidence":"ChIP-PCR and dual-luciferase for PRRX1 binding, MS and Co-IP for EGFR-PKCα complex, and xenograft model","pmids":["39528080"],"confidence":"Medium","gaps":["Generality of PRRX1 control of ANXA6 beyond bladder cancer untested","Does not address how SUMOylation and PRRX1 regulation interrelate"]},{"year":2025,"claim":"Resolved the metabolic phenotype to a specific defect, showing ANXA6 is required for alanine-dependent hepatic gluconeogenesis and fasting glucose maintenance.","evidence":"Anxa6-/- mice with calorimetry, tolerance tests, and expression profiling of Gpt2/Ldha2/SNAT4","pmids":["40802799"],"confidence":"Medium","gaps":["Mechanism linking ANXA6 to alanine-handling gene expression not defined","Whether the effect is cell-autonomous in hepatocytes vs systemic is unclear"]},{"year":2025,"claim":"Connected ANXA6 to innate-immune and ferroptosis control by showing it promotes K63-linked ubiquitination of cGAS to restrain cGAS-STING signaling.","evidence":"Co-localization, ubiquitination assays, cGAS-STING pathway blotting, and ROS/lipid-peroxide measurements after ANXA6 knockdown in cervical cancer cells","pmids":["41219880"],"confidence":"Medium","gaps":["The E3 ligase ANXA6 recruits to cGAS is not identified","Whether ANXA6 binds cGAS directly is not established"]},{"year":2025,"claim":"Implicated ANXA6 in macrophage inflammatory programming at the maternal-fetal interface via NF-κB/NLRP3-driven pyroptosis.","evidence":"ANXA6 overexpression in primary decidual macrophages with ROS/membrane-potential assays, NLRP3-inhibitor rescue, and trophoblast co-culture","pmids":["40959268"],"confidence":"Medium","gaps":["Direct molecular trigger upstream of NF-κB activation by ANXA6 unknown","Endogenous relevance versus overexpression artifact not separated"]},{"year":null,"claim":"How ANXA6's Ca2+-dependent membrane binding mechanistically unifies its disparate roles in EGFR scaffolding, autophagic trafficking, ubiquitin-mediated immune control, and hepatic metabolism remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model linking lipid binding to partner recruitment","Direct enzymatic or adaptor activity not biochemically reconstituted","Whether SUMOylation gates membrane translocation across contexts is untested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[6]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[2,11]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,12]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6,15]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[0,1,10]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,11]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[8,12]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[4,5]}],"complexes":[],"partners":["EGFR","PKCA","TRPV2","SNAP23","CGAS","SUMO1","MG53"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P08133","full_name":"Annexin A6","aliases":["67 kDa calelectrin","Annexin VI","Annexin-6","Calphobindin-II","CPB-II","Chromobindin-20","Lipocortin VI","Protein III","p68","p70"],"length_aa":673,"mass_kda":75.9,"function":"May associate with CD21. May regulate the release of Ca(2+) from intracellular stores","subcellular_location":"Cytoplasm; Melanosome","url":"https://www.uniprot.org/uniprotkb/P08133/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ANXA6","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CALD1","stoichiometry":0.2},{"gene":"CEP192","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/ANXA6","total_profiled":1310},"omim":[{"mim_id":"114070","title":"ANNEXIN A6; ANXA6","url":"https://www.omim.org/entry/114070"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Basal body","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ANXA6"},"hgnc":{"alias_symbol":[],"prev_symbol":["ANX6"]},"alphafold":{"accession":"P08133","domains":[{"cath_id":"1.10.220.10","chopping":"21-90","consensus_level":"medium","plddt":97.386,"start":21,"end":90},{"cath_id":"1.10.220.10","chopping":"252-347","consensus_level":"medium","plddt":92.1549,"start":252,"end":347},{"cath_id":"1.10.220.10","chopping":"364-421","consensus_level":"medium","plddt":97.4493,"start":364,"end":421},{"cath_id":"1.10.220.10","chopping":"435-505","consensus_level":"medium","plddt":97.5837,"start":435,"end":505},{"cath_id":"1.10.220.10","chopping":"515-597","consensus_level":"medium","plddt":88.8701,"start":515,"end":597},{"cath_id":"1.10.220.10","chopping":"599-668","consensus_level":"medium","plddt":96.7981,"start":599,"end":668}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P08133","model_url":"https://alphafold.ebi.ac.uk/files/AF-P08133-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P08133-F1-predicted_aligned_error_v6.png","plddt_mean":94.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ANXA6","jax_strain_url":"https://www.jax.org/strain/search?query=ANXA6"},"sequence":{"accession":"P08133","fasta_url":"https://rest.uniprot.org/uniprotkb/P08133.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P08133/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P08133"}},"corpus_meta":[{"pmid":"32373608","id":"PMC_32373608","title":"ANXA6 Contributes to Radioresistance by Promoting Autophagy via Inhibiting the PI3K/AKT/mTOR Signaling Pathway in Nasopharyngeal Carcinoma.","date":"2020","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/32373608","citation_count":41,"is_preprint":false},{"pmid":"33135350","id":"PMC_33135350","title":"ANXA6 suppresses the tumorigenesis of cervical cancer through autophagy induction.","date":"2020","source":"Clinical and translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33135350","citation_count":34,"is_preprint":false},{"pmid":"34676207","id":"PMC_34676207","title":"Breast Cancer Stem Cell-Derived ANXA6-Containing Exosomes Sustain Paclitaxel Resistance and Cancer Aggressiveness in Breast Cancer.","date":"2021","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/34676207","citation_count":28,"is_preprint":false},{"pmid":"35456441","id":"PMC_35456441","title":"miR-24-3p Dominates the Proliferation and Differentiation of Chicken Intramuscular Preadipocytes by Blocking ANXA6 Expression.","date":"2022","source":"Genes","url":"https://pubmed.ncbi.nlm.nih.gov/35456441","citation_count":26,"is_preprint":false},{"pmid":"23541940","id":"PMC_23541940","title":"TNIP1/ANXA6 and CSMD1 variants interacting with cigarette smoking, alcohol intake affect risk of psoriasis.","date":"2013","source":"Journal of dermatological science","url":"https://pubmed.ncbi.nlm.nih.gov/23541940","citation_count":26,"is_preprint":false},{"pmid":"37129645","id":"PMC_37129645","title":"ANXA6: a key molecular player in cancer progression and drug resistance.","date":"2023","source":"Discover oncology","url":"https://pubmed.ncbi.nlm.nih.gov/37129645","citation_count":22,"is_preprint":false},{"pmid":"30110341","id":"PMC_30110341","title":"Altered hepatic glucose homeostasis in AnxA6-KO mice fed a high-fat diet.","date":"2018","source":"PloS 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biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/39577524","citation_count":6,"is_preprint":false},{"pmid":"23360953","id":"PMC_23360953","title":"Interaction of AnxA6 with isolated and artificial lipid microdomains; importance of lipid composition and calcium content.","date":"2013","source":"Molecular bioSystems","url":"https://pubmed.ncbi.nlm.nih.gov/23360953","citation_count":6,"is_preprint":false},{"pmid":"39528080","id":"PMC_39528080","title":"Transcription factor PRRX1-activated ANXA6 facilitates EGFR-PKCα complex formation and enhances cisplatin sensitivity in bladder cancer.","date":"2024","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39528080","citation_count":4,"is_preprint":false},{"pmid":"37311953","id":"PMC_37311953","title":"Comprehensive analysis the diagnosis, malignant progression and immune infiltrate of ANXA6 in prostate cancer.","date":"2023","source":"Genes & 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longevity","url":"https://pubmed.ncbi.nlm.nih.gov/38188996","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.09.23.677988","title":"Duchenne muscular dystrophy is driven by defective membrane repair and annexin-A2 dysregulation in skeletal muscle","date":"2025-09-23","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.23.677988","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.01.24.634638","title":"Fasting-induced hepatic gluconeogenesis is compromised in  <i>  Anxa6  <sup>−/−</sup>  </i>  mice","date":"2025-01-26","source":"bioRxiv","url":"https://doi.org/10.1101/2025.01.24.634638","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.06.15.659434","title":"A map of Non-translated RNA (nt-RNA) junctions in cancer genomes: a database resource of unproductive 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cells","date":"2024-10-25","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.22.619710","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":24088,"output_tokens":4571,"usd":0.070415,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12655,"output_tokens":3899,"usd":0.080375,"stage2_stop_reason":"end_turn"},"total_usd":0.15079,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2020,\n      \"finding\": \"ANXA6 promotes autophagy by inhibiting the PI3K/AKT/mTOR signaling pathway in nasopharyngeal carcinoma cells; siRNA knockdown of ANXA6 activated PI3K/AKT/mTOR and suppressed autophagy, and co-treatment with PI3K/AKT/mTOR inhibitor CAL101 reversed siANXA6-reduced autophagy, establishing negative regulatory feedback.\",\n      \"method\": \"siRNA knockdown, pharmacological inhibition (CAL101), western blotting of pathway components\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined molecular phenotype and pharmacological rescue, single lab, two orthogonal perturbation methods\",\n      \"pmids\": [\"32373608\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ANXA6 enables ATG9A-positive vesicle trafficking from recycling endosomes to autophagosomes through RAB GTPases and F-actin, thereby facilitating autophagosome formation; ANXA6 also suppresses mTOR activity by inhibiting PI3K-AKT and ERK signaling pathways.\",\n      \"method\": \"siRNA knockdown, re-expression rescue, GO analysis of interactors, confocal imaging of ATG9A vesicle trafficking, western blotting of mTOR pathway\",\n      \"journal\": \"Clinical and translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function plus rescue with defined trafficking phenotype, single lab, multiple orthogonal readouts\",\n      \"pmids\": [\"33135350\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ANXA6 is SUMOylated by SUMO1 at lysine K299; this SUMOylation promotes ANXA6 binding to EGFR upon EGF stimulation and facilitates EGFR-PKCα complex formation, which decreases phosphorylation of EGFR and ERK1/2. The K299R mutant (SUMO-deficient) loses the ability to suppress EGFR-ERK signaling and confers drug resistance to gefitinib.\",\n      \"method\": \"Co-transfection with SUMO1/UBC9/His-SUMO1 plasmids, immunoprecipitation, LC-MS/MS identification of SUMOylation sites, site-directed mutagenesis (K299R), shRNA knockdown, lentiviral overexpression, xenograft mouse model\",\n      \"journal\": \"Cell communication and signaling : CCS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — SUMOylation site identified by MS and validated by mutagenesis, complex formation confirmed by Co-IP, functional consequence tested in vitro and in vivo\",\n      \"pmids\": [\"37528485\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"The EHEC effector protein EspF interacts with host protein ANXA6 through EspF's C-terminal domain; this EspF-ANXA6 interaction activates MLCK, induces MLC and PKCα phosphorylation, downregulates calmodulin, and disrupts tight junction proteins ZO-1 and occludin, leading to intestinal barrier disruption.\",\n      \"method\": \"FRET, co-immunoprecipitation, domain mapping, western blotting, immunofluorescence of tight junction distribution\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — interaction confirmed by two orthogonal methods (FRET + Co-IP), downstream pathway assessed by western blot, single lab\",\n      \"pmids\": [\"33425920\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"AnxA6-knockout mice on a high-fat diet show increased insulin-stimulated Akt and S6K phosphorylation in liver but fail to suppress hepatic gluconeogenesis upon insulin exposure; AnxA6-depleted HuH7 hepatocytes also fail to reduce glucose production upon insulin, implicating AnxA6 in fine-tuning hepatic glucose metabolism downstream of or parallel to insulin signaling.\",\n      \"method\": \"AnxA6-KO mouse model, OGTT, ITT, western blotting of phospho-Akt and phospho-S6, glucose production assay in AnxA6-depleted hepatocytes\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse phenotype corroborated by in vitro hepatocyte loss-of-function, multiple metabolic readouts, single lab\",\n      \"pmids\": [\"30110341\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Anxa6-knockout mice display rapid fasting hypoglycaemia due to inability to utilize alanine for hepatic gluconeogenesis (glucose-alanine cycle impairment), associated with reduced expression of hepatic alanine aminotransferase 2 (Gpt2), lactate dehydrogenase (Ldha2), and the alanine transporter SNAT4.\",\n      \"method\": \"Anxa6-/- mouse model, indirect calorimetry, glucose/insulin/alanine tolerance tests, biochemical analysis, gene expression profiling\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse with multiple metabolic tolerance tests and molecular pathway readouts, single lab, peer-reviewed\",\n      \"pmids\": [\"40802799\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"In Niemann-Pick type C disease cells, AnxA6 translocates to cholesterol- and sphingomyelin-enriched lipid microdomains (DRMs) upon a rise in Ca2+ concentration; in vitro Langmuir monolayer experiments showed Ca2+ is the primary factor regulating AnxA6 interaction with DPPC/sphingomyelin monolayers and its localization in these microdomains.\",\n      \"method\": \"Lipid microdomain (DRM) isolation by fractionation, Langmuir monolayer assay with defined lipid compositions, Ca2+ titration\",\n      \"journal\": \"Molecular bioSystems\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro reconstitution with Langmuir monolayers plus cellular fractionation, single lab, two orthogonal methods\",\n      \"pmids\": [\"23360953\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"EZH2 suppresses ANXA6 expression by increasing H3K27me3 histone methylation at the ANXA6 promoter; overexpression of EZH2 reduced ANXA6 levels, inhibited Ang II-induced ROS, and protected against vascular smooth muscle cell senescence, while co-overexpression of ANXA6 partially reversed the EZH2 protective effect.\",\n      \"method\": \"ChIP (H3K27me3 at ANXA6 promoter), lentiviral overexpression of EZH2 and ANXA6, SA-β-gal staining, flow cytometry, ROS assay, western blotting\",\n      \"journal\": \"Oxidative medicine and cellular longevity\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 2 / Weak — ChIP validates epigenetic mechanism but paper was subsequently retracted (PMID:38188996); evidence is unreliable\",\n      \"pmids\": [\"36160712\", \"38188996\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CRISPR-Cas9 knockout of ANXA6 in HEK293FT cells significantly increased influenza A virus replication and viral titer, demonstrating that AnxA6 acts as a host restriction factor counteracting influenza virion packaging.\",\n      \"method\": \"CRISPR-Cas9 gene knockout, TCID50 assay, flow cytometry, viral growth curve experiments\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO with defined functional phenotype (viral restriction), two quantitative methods, single lab\",\n      \"pmids\": [\"34673207\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Apigenin disrupts intracellular distribution of AnxA6 (blocking AnxA6 aggregation) and TNAP attachment to membranes in osteoblast cell lines, thereby modulating matrix vesicle-mediated mineralization; the effects were cell-type dependent across hFOB 1.19, Saos-2, and HCASMC cells.\",\n      \"method\": \"Fluorescence microscopy of AnxA6 distribution, TEM-gold immunolabeling of vesicles, AR-S/CPC mineralization assay, ELISA for TNAP activity\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — localization experiments with functional correlation but no direct mechanistic perturbation of AnxA6, single lab\",\n      \"pmids\": [\"36361965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ANXA6 interacts with TRPV2; ANXA6 induces autophagy in head and neck squamous cell carcinoma by inhibiting the AKT/mTOR signaling pathway, and TRPV2 inhibition reverses ANXA6-induced autophagy and lymphatic metastasis, placing TRPV2 downstream of ANXA6 in this pathway.\",\n      \"method\": \"ANXA6 overexpression/knockdown in vitro and in vivo (lymphatic metastasis model), Co-IP/molecular interaction assays, AKT/mTOR pathway western blotting, TRPV2 pharmacological inhibition\",\n      \"journal\": \"Experimental hematology & oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — interaction shown with Co-IP, epistasis demonstrated by pharmacological rescue, in vivo model, single lab\",\n      \"pmids\": [\"37138336\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PRRX1 transcription factor directly promotes ANXA6 transcription (validated by ChIP-PCR and dual-luciferase assay); ANXA6 in turn promotes PKCα/EGFR complex formation by co-immunoprecipitation, inhibiting EGFR phosphorylation and downstream AKT and ERK1/2, thereby enhancing cisplatin sensitivity in bladder cancer cells.\",\n      \"method\": \"ChIP-PCR, dual-luciferase reporter assay, mass spectrometry, immunofluorescence, co-immunoprecipitation, xenograft model\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — transcription factor binding validated by ChIP and luciferase; EGFR-PKCα complex by Co-IP and MS; single lab with multiple methods\",\n      \"pmids\": [\"39528080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ANXA6 co-localizes with cGAS and promotes ubiquitination (K63-linked) of cGAS, suppressing the cGAS-STING signaling pathway; ANXA6 knockdown inhibits K63-linked ubiquitination of cGAS, leading to cGAS accumulation, activation of phospho-TBK1/IRF3, type I IFN-stimulated genes, increased ROS and lipid peroxides, and ultimately ferroptosis in cervical cancer cells.\",\n      \"method\": \"Co-localization (immunofluorescence), western blotting of cGAS-STING pathway components, ubiquitination assay, ROS and lipid peroxide measurements, ANXA6 knockdown\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-localization and ubiquitination assay establish mechanism, pathway validated by downstream signaling, single lab\",\n      \"pmids\": [\"41219880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MG53 (mitsugumin 53) binds AnxA6 and exerts cytoprotective effects on iPSCs under hypoxia/reoxygenation injury; overexpression of AnxA6 abolishes MG53's membrane-stabilizing cytoprotective effects, indicating AnxA6 negatively modulates MG53-mediated membrane repair.\",\n      \"method\": \"Co-immunoprecipitation (MG53-AnxA6 binding), AnxA6 overexpression rescue experiment, cell viability assays, in vivo iPSC transplantation in MIRI model\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — binding shown by Co-IP but mechanistic detail is limited; single lab, limited orthogonal validation\",\n      \"pmids\": [\"41106752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ANXA6 overexpression in decidual macrophages promotes mitochondrial ROS accumulation and reduces mitochondrial membrane potential via the NF-κB signaling pathway, triggering NLRP3-dependent pyroptosis and M1 macrophage polarization; conditioned medium from ANXA6-overexpressing macrophages inhibits trophoblast function via TNF-α-mediated suppression of the PI3K/AKT axis.\",\n      \"method\": \"ANXA6 overexpression in primary macrophages, ROS assay, mitochondrial membrane potential assay, NLRP3 inhibitor rescue, NF-κB pathway analysis, in vitro trophoblast co-culture\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss/gain of function with defined molecular pathway, pharmacological rescue with NLRP3 inhibitor and ROS scavenger, single lab\",\n      \"pmids\": [\"40959268\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"AnxA6 interacts with SNAP23 and this interaction is required for the translocation of AnxA6 to cellular membranes, which facilitates secretion of small extracellular vesicles (EVs) and pro-inflammatory cytokines (MCP-1/CCL2, IL-8) in triple-negative breast cancer cells; AnxA6 downregulation strongly reduces EV secretion and associated cholesterol and MCP-1 content.\",\n      \"method\": \"Co-immunoprecipitation (AnxA6-SNAP23), cytokine arrays of secreted factors, EV isolation and characterization, AnxA6 knockdown and overexpression, cholesterol quantification in EVs\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — Co-IP interaction plus functional secretion data, preprint not yet peer-reviewed, single lab\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In healthy skeletal muscle cells, mechanical stress induces upregulation of ANXA6 (and ANXA5), suggesting ANXA6 participates in an adaptive membrane repair response; in Duchenne muscular dystrophy (DMD) cells, ANXA6 expression is dysregulated, contributing to defective membrane repair.\",\n      \"method\": \"Live-cell imaging of GFP-tagged annexins, shear stress and laser irradiation injury assays, western blotting, immunohistochemistry of DMD patient biopsies\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — localization and upregulation data from injury assays establish correlation with membrane repair; preprint, mechanistic detail for ANXA6 specifically is limited\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"ANXA6 (Annexin A6) is a Ca2+-dependent membrane- and phospholipid-binding scaffold protein that: (1) suppresses EGFR-ERK/AKT signaling by facilitating EGFR-PKCα complex formation (a function potentiated by SUMO1 modification at K299); (2) promotes autophagy by inhibiting PI3K/AKT/mTOR signaling and facilitating ATG9A vesicle trafficking via RAB GTPases and F-actin; (3) regulates cholesterol homeostasis by Ca2+-dependent translocation into lipid microdomains; (4) acts as a host restriction factor for influenza replication; (5) suppresses cGAS-STING signaling by promoting K63-linked ubiquitination of cGAS; (6) is required for hepatic alanine-dependent gluconeogenesis; and (7) facilitates secretion of extracellular vesicles and pro-inflammatory cytokines through interaction with SNAP23.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ANXA6 is a Ca2+-responsive membrane- and lipid-associated scaffold protein that integrates growth-factor signaling, autophagy, and membrane organization across diverse cell types [#1, #6]. Its best-characterized signaling output is suppression of the EGFR axis: ANXA6 promotes formation of an EGFR-PKC\\u03b1 complex that lowers EGFR and ERK1/2 phosphorylation, a function potentiated by SUMO1 conjugation at lysine K299, with the SUMO-deficient K299R mutant losing this activity and conferring gefitinib resistance [#2]. The same EGFR-PKC\\u03b1-restraining activity is engaged downstream of the transcription factor PRRX1 to enhance cisplatin sensitivity in bladder cancer [#11]. ANXA6 also drives autophagy by inhibiting PI3K/AKT/mTOR (and ERK) signaling and by enabling ATG9A-vesicle trafficking from recycling endosomes to autophagosomes via RAB GTPases and F-actin [#0, #1], with TRPV2 acting downstream of ANXA6-induced autophagy [#10]. Consistent with Ca2+-dependent lipid binding, ANXA6 translocates into cholesterol- and sphingomyelin-rich microdomains as intracellular Ca2+ rises [#6], and supports SNAP23-dependent membrane translocation that facilitates secretion of extracellular vesicles and pro-inflammatory cytokines [#15]. Beyond these roles, ANXA6 acts as a host restriction factor limiting influenza A virus replication [#8], suppresses cGAS-STING signaling by promoting K63-linked ubiquitination of cGAS [#12], and is required in liver for insulin-modulated suppression of hepatic gluconeogenesis and for alanine-dependent gluconeogenesis [#4, #5].\",\n  \"teleology\": [\n    {\n      \"year\": 2013,\n      \"claim\": \"Established the biophysical basis for ANXA6 membrane targeting by showing that Ca2+ drives its translocation into cholesterol/sphingomyelin-enriched microdomains.\",\n      \"evidence\": \"DRM fractionation in Niemann-Pick type C cells plus Langmuir monolayer reconstitution with Ca2+ titration\",\n      \"pmids\": [\"23360953\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not identify the protein partners ANXA6 recruits to these microdomains\", \"Functional consequence for cholesterol handling in vivo not resolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Connected ANXA6 to systemic metabolism by showing knockout livers fail to suppress gluconeogenesis upon insulin, placing ANXA6 in the hepatic insulin response.\",\n      \"evidence\": \"AnxA6-KO mice with OGTT/ITT and phospho-Akt/S6 blotting, plus glucose-production assays in depleted hepatocytes\",\n      \"pmids\": [\"30110341\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular target through which ANXA6 modulates Akt/S6K not defined\", \"Direct binding partners in hepatocytes not identified\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined ANXA6 as a positive regulator of autophagy operating both by inhibiting PI3K/AKT/mTOR and by physically enabling ATG9A vesicle delivery to autophagosomes.\",\n      \"evidence\": \"siRNA knockdown with re-expression rescue, pharmacological PI3K inhibition, and confocal imaging of ATG9A trafficking\",\n      \"pmids\": [\"32373608\", \"33135350\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ANXA6 binds ATG9A directly or acts through RAB/F-actin scaffolds is unresolved\", \"How the mTOR-inhibitory and trafficking functions are coordinated is unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified ANXA6 as a host target of the EHEC effector EspF, linking it to tight-junction integrity via MLCK/PKC\\u03b1 signaling.\",\n      \"evidence\": \"FRET and Co-IP with domain mapping, plus tight-junction protein immunofluorescence\",\n      \"pmids\": [\"33425920\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not establish ANXA6's endogenous role at tight junctions absent infection\", \"Direct enzymatic consequence of EspF binding to ANXA6 unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Showed ANXA6 functions as a host restriction factor against influenza A virus.\",\n      \"evidence\": \"CRISPR-Cas9 knockout in HEK293FT with TCID50 and viral growth-curve readouts\",\n      \"pmids\": [\"34673207\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Step of the viral life cycle ANXA6 acts on is not mechanistically pinned\", \"No viral or host binding partner identified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealed that SUMO1 modification at K299 is required for ANXA6 to suppress EGFR-ERK signaling, mechanistically tying a post-translational modification to drug response.\",\n      \"evidence\": \"MS-mapped SUMOylation site validated by K299R mutagenesis, Co-IP of EGFR-PKC\\u03b1 complex, and xenograft model\",\n      \"pmids\": [\"37528485\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The SUMO E3 ligase and conditions controlling K299 modification are not defined\", \"Structural basis of SUMO-dependent EGFR binding unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended the autophagy axis by identifying TRPV2 as a downstream effector of ANXA6-induced, AKT/mTOR-dependent autophagy in head and neck cancer metastasis.\",\n      \"evidence\": \"Gain/loss-of-function in vitro and in a lymphatic metastasis model, Co-IP, and TRPV2 pharmacological rescue\",\n      \"pmids\": [\"37138336\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ANXA6-TRPV2 binding is direct and Ca2+-dependent is unresolved\", \"Reciprocal validation of the interaction is limited\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placed the EGFR-suppressive ANXA6 function within a transcriptional circuit, showing PRRX1 directly drives ANXA6 expression to enhance chemosensitivity.\",\n      \"evidence\": \"ChIP-PCR and dual-luciferase for PRRX1 binding, MS and Co-IP for EGFR-PKC\\u03b1 complex, and xenograft model\",\n      \"pmids\": [\"39528080\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Generality of PRRX1 control of ANXA6 beyond bladder cancer untested\", \"Does not address how SUMOylation and PRRX1 regulation interrelate\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Resolved the metabolic phenotype to a specific defect, showing ANXA6 is required for alanine-dependent hepatic gluconeogenesis and fasting glucose maintenance.\",\n      \"evidence\": \"Anxa6-/- mice with calorimetry, tolerance tests, and expression profiling of Gpt2/Ldha2/SNAT4\",\n      \"pmids\": [\"40802799\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking ANXA6 to alanine-handling gene expression not defined\", \"Whether the effect is cell-autonomous in hepatocytes vs systemic is unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected ANXA6 to innate-immune and ferroptosis control by showing it promotes K63-linked ubiquitination of cGAS to restrain cGAS-STING signaling.\",\n      \"evidence\": \"Co-localization, ubiquitination assays, cGAS-STING pathway blotting, and ROS/lipid-peroxide measurements after ANXA6 knockdown in cervical cancer cells\",\n      \"pmids\": [\"41219880\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"The E3 ligase ANXA6 recruits to cGAS is not identified\", \"Whether ANXA6 binds cGAS directly is not established\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Implicated ANXA6 in macrophage inflammatory programming at the maternal-fetal interface via NF-\\u03baB/NLRP3-driven pyroptosis.\",\n      \"evidence\": \"ANXA6 overexpression in primary decidual macrophages with ROS/membrane-potential assays, NLRP3-inhibitor rescue, and trophoblast co-culture\",\n      \"pmids\": [\"40959268\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular trigger upstream of NF-\\u03baB activation by ANXA6 unknown\", \"Endogenous relevance versus overexpression artifact not separated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How ANXA6's Ca2+-dependent membrane binding mechanistically unifies its disparate roles in EGFR scaffolding, autophagic trafficking, ubiquitin-mediated immune control, and hepatic metabolism remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model linking lipid binding to partner recruitment\", \"Direct enzymatic or adaptor activity not biochemically reconstituted\", \"Whether SUMOylation gates membrane translocation across contexts is untested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [2, 11]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 12]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6, 15]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [0, 1, 10]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 11]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [8, 12]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [4, 5]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"EGFR\", \"PKCA\", \"TRPV2\", \"SNAP23\", \"CGAS\", \"SUMO1\", \"MG53\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}