{"gene":"ANXA6","run_date":"2026-04-28T17:12:37","timeline":{"discoveries":[{"year":1988,"finding":"The primary amino acid sequence of human p68 (later named ANXA6) was determined from cDNA clones, revealing it is a membrane-associated Ca2+-binding protein with an internally repetitive structure comprising eight repeats, each containing a conserved sequence motif. It was identified as a member of the annexin family, related to p36, lipocortin I, and protein II, and the cDNA data suggested alternatively spliced mRNAs could encode different p68 protein species.","method":"cDNA cloning, sequencing, and sequence analysis","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 — original structural characterization by cDNA sequencing with functional inference; foundational paper (161 citations)","pmids":["3258820"],"is_preprint":false},{"year":1990,"finding":"ANXA6 (referred to as p68) was shown to inhibit phospholipid vesicle membrane fusion at threshold Ca2+ concentrations, distinguishing it functionally from other annexins. It binds Ca2+-dependently to phosphatidic acid, phosphatidylserine, phosphatidylinositol, and phosphatidylethanolamine vesicles, but not phosphatidylcholine vesicles. These quantitative and qualitative differences among annexins suggest distinct intracellular functions.","method":"In vitro phospholipid vesicle binding assay, vesicle aggregation and fusion assays with defined lipid compositions and Ca2+ concentrations","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1 — direct in vitro reconstitution with quantitative lipid binding and fusion assays; highly cited (276 citations)","pmids":["2138016"],"is_preprint":false},{"year":2013,"finding":"In Niemann-Pick type C (NPC) disease cells, AnxA6 translocates to lipid microdomains (detergent-resistant membranes, DRMs) enriched in cholesterol and sphingomyelin upon a rise in Ca2+ concentration. In vitro Langmuir monolayer experiments showed that Ca2+ is the primary factor regulating AnxA6 interaction with neutral lipid monolayers (DPPC and sphingomyelin), indicating Ca2+-driven partitioning of AnxA6 into cholesterol/sphingomyelin-enriched microdomains contributes to NPC pathology.","method":"Lipid microdomain (DRM) isolation and fractionation, Langmuir monolayer in vitro lipid-protein interaction assays, calcium titration","journal":"Molecular bioSystems","confidence":"Medium","confidence_rationale":"Tier 2 — cell fractionation combined with Tier 1 in vitro reconstitution; single lab","pmids":["23360953"],"is_preprint":false},{"year":2016,"finding":"Cancer-associated fibroblasts (CAFs) secrete ANXA6-containing extracellular vesicles (EVs) that carry an ANXA6/LRP1/TSP1 complex. Formation of this complex is restricted to CAFs under physiopathologic conditions. Tumor cell uptake of CAF-derived ANXA6+ EVs promotes pancreatic ductal adenocarcinoma (PDA) cell survival and migration. Depletion of ANXA6 in CAFs impaired complex formation and reduced PDA aggressiveness and metastasis in vivo, while injection of ANXA6+ EVs enhanced tumorigenesis.","method":"Proteomic stromal signature analysis, co-immunoprecipitation to identify the ANXA6/LRP1/TSP1 complex, ANXA6 depletion in CAFs, in vivo xenograft and metastasis assays, EV uptake experiments","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, in vivo rescue experiments, multiple orthogonal methods; highly cited (182 citations)","pmids":["27701147"],"is_preprint":false},{"year":2017,"finding":"AnxA6 was identified as the target antigen of the inhibitory monoclonal antibody MAb 9E1 using immunoprecipitation/LC-MS/MS. Silencing of AnxA6 markedly reduced the invasive capacity of pancreatic and lung squamous cancer cells in vitro, identifying AnxA6 as a functionally relevant pro-invasive target at the cancer cell surface.","method":"Immunoprecipitation/LC-MS/MS target identification, AnxA6 siRNA knockdown, invasion assays in vitro","journal":"British journal of cancer","confidence":"Medium","confidence_rationale":"Tier 2 — MS-based target identification combined with functional KD; single lab","pmids":["28881357"],"is_preprint":false},{"year":2018,"finding":"AnxA6-knockout (KO) mice fed a high-fat diet gained less weight and showed reduced adiposity compared to wild-type controls. AnxA6-KO mice displayed increased insulin-inducible phosphorylation of Akt and S6K in liver, but failed to downregulate hepatic gluconeogenesis despite normal insulin signaling. AnxA6-depleted HuH7 hepatocytes were unable to reduce glucose production upon insulin exposure, and KO animals showed increased hepatic glycogen storage, demonstrating that AnxA6 contributes to fine-tuning of hepatic glucose metabolism.","method":"AnxA6-KO mouse model, high-fat diet feeding, oral glucose tolerance tests (OGTT), insulin tolerance tests (ITT), Western blot for Akt/S6K phosphorylation, glucose production assay in AnxA6-depleted hepatocytes","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 — clean KO mouse with defined metabolic phenotypes, confirmed in vitro in hepatocytes with multiple readouts","pmids":["30110341"],"is_preprint":false},{"year":2020,"finding":"ANXA6 promotes autophagy by inhibiting the PI3K/AKT/mTOR signaling pathway in nasopharyngeal carcinoma cells, thereby contributing to radioresistance. ANXA6 siRNA suppressed autophagy through activation (de-repression) of PI3K/AKT/mTOR, and the PI3K inhibitor CAL101 reversed siANXA6-reduced autophagy. A negative feedback loop was identified in which suppression of PI3K/AKT/mTOR by CAL101 increased ANXA6 expression.","method":"ANXA6 siRNA knockdown, LC3 siRNA knockdown, PI3K/AKT/mTOR inhibitor (CAL101) treatment, autophagy assays, Western blot for pathway components","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological and genetic epistasis with defined pathway readouts; single lab","pmids":["32373608"],"is_preprint":false},{"year":2020,"finding":"EHEC effector protein EspF physically interacts with host protein ANXA6 through its C-terminal domain, as verified by FRET and co-immunoprecipitation. Co-expression of EspF-ANXA6 activated myosin light chain kinase (MLCK), induced phosphorylation of myosin light chain (MLC) and PKCα, downregulated Calmodulin, and decreased levels and disrupted distribution of tight junction proteins ZO-1 and occludin, revealing a mechanism by which bacterial infection perturbs epithelial barrier function through ANXA6.","method":"FRET, co-immunoprecipitation, domain mapping, Western blot for MLCK/MLC/PKCα phosphorylation and tight junction proteins, confocal microscopy for ZO-1 distribution","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 — FRET plus co-IP for interaction, combined with downstream pathway readouts; single lab","pmids":["33425920"],"is_preprint":false},{"year":2020,"finding":"ANXA6 enables appropriate ATG9A+ vesicle trafficking from endosomes to autophagosomes through RAB GTPases and/or F-actin, thereby facilitating autophagosome formation. ANXA6 knockdown attenuated starvation-induced autophagy, while restoration of expression enhanced it. ANXA6 also suppresses mTOR activity through inhibition of PI3K-AKT and ERK signaling pathways, providing dual control of autophagy.","method":"ANXA6 siRNA knockdown, ANXA6 rescue overexpression, GO analysis of ANXA6 interaction targets, ATG9A vesicle trafficking assays, mTOR/PI3K-AKT/ERK signaling Western blot, LC3 reporter assays","journal":"Clinical and translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 — KD/rescue with multiple readouts; GO-based interactome inference supports mechanism; single lab","pmids":["33135350"],"is_preprint":false},{"year":2021,"finding":"ANXA6-containing exosomes (ANXA6-exo) derived from breast cancer stem cells promote paclitaxel resistance and cancer progression in recipient cells via autophagy induction in a YAP1-dependent manner. ANXA6-exo upregulated YAP1 to dysregulate the Hippo pathway, and silencing YAP1 abrogated the resistance effects. Autophagy inhibition with 3-methyladenine blocked ANXA6-exo-mediated PTX resistance.","method":"Exosome isolation and transfer experiments, ANXA6 and YAP1 siRNA knockdown, autophagy inhibitor (3-methyladenine), paclitaxel resistance assays, stem cell enrichment assays","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 — exosome transfer with genetic and pharmacological epistasis; single lab","pmids":["34676207"],"is_preprint":false},{"year":2021,"finding":"CRISPR-Cas9 knockout of the ANXA6 gene in HEK293FT cells significantly increased influenza A virus replication and accumulation, demonstrating that endogenous ANXA6 functions as a host restriction factor that counteracts influenza virion packaging.","method":"CRISPR-Cas9 gene knockout, TCID50 viral titer assay, flow cytometry, viral growth curve experiments in ANXA6-/- versus parental cells","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 2 — clean CRISPR KO with defined viral replication phenotype; single lab","pmids":["34673207"],"is_preprint":false},{"year":2022,"finding":"In osteoblasts, AnxA6 plays a role in matrix vesicle-mediated mineralization. Apigenin treatment disrupted intracellular distribution of AnxA6 by blocking AnxA6 aggregation and TNAP attachment to membranes, and modulated TNAP enzymatic activity, affecting mineral composition and structure. AnxA6 distribution was examined by fluorescence microscopy and TEM-gold in cells and vesicles.","method":"Fluorescence microscopy (FM), TEM-gold labeling of vesicles, ELISA for TNAP activity, AR-S/CPC mineralization assay, apigenin pharmacological treatment in hFOB1.19, Saos-2, and HCASMC cell lines","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization by TEM-gold and FM tied to functional consequence (mineralization); single lab","pmids":["36361965"],"is_preprint":false},{"year":2022,"finding":"miR-24-3p directly binds the 3'UTR of ANXA6 mRNA and suppresses ANXA6 expression, thereby promoting proliferation and inhibiting adipogenic differentiation of chicken intramuscular preadipocytes. Overexpression of ANXA6 inhibited proliferation and promoted differentiation, while ANXA6 knockdown had opposite effects. Rescue with an ANXA6 overexpression vector eliminated the impact of miR-24-3p mimics, confirming ANXA6 as the functional mediator.","method":"Dual-luciferase reporter assay for miR-24-3p binding to ANXA6 3'UTR, ANXA6 overexpression and knockdown, miR-24-3p mimics/inhibitors, cell proliferation and adipogenic differentiation assays","journal":"Genes","confidence":"Medium","confidence_rationale":"Tier 2 — dual-luciferase validates direct miRNA-target interaction; rescue experiment confirms functional axis; single lab","pmids":["35456441"],"is_preprint":false},{"year":2023,"finding":"AnxA6 is SUMOylated by SUMO1, with K299 identified as a key SUMOylation site. SUMOylated AnxA6 preferentially binds EGFR upon EGF stimulation and facilitates EGFR-PKCα complex formation, which decreases EGF-induced phosphorylation of EGFR and ERK1/2 and reduces cyclin D1 expression. The SUMO-site mutant AnxA6K299R showed reduced suppression of EGFR-ERK1/2 phosphorylation, reduced tumor suppression, and conferred gefitinib resistance. This mechanism also operates on mutant EGFR (T790M/L858R).","method":"Co-transfection with SUMO1/UBC9 plasmids, immunoprecipitation for SUMOylated AnxA6, LC-MS/MS for SUMOylation site mapping, site-directed mutagenesis (K299R), shRNA knockdown, lentiviral overexpression, Western blot, CCK8/colony/wound healing assays, xenograft nude mouse model","journal":"Cell communication and signaling","confidence":"High","confidence_rationale":"Tier 1-2 — SUMOylation site identified by MS and validated by mutagenesis, functional consequences confirmed in vitro and in vivo; multiple orthogonal methods","pmids":["37528485"],"is_preprint":false},{"year":2023,"finding":"The ANXA6/TRPV2 axis promotes lymphatic metastasis in head and neck squamous cell carcinoma by inducing autophagy through inhibition of the AKT/mTOR signaling pathway. ANXA6 expression positively correlated with TRPV2 expression, and TRPV2 inhibition reversed ANXA6-induced autophagy and lymphatic metastasis, placing TRPV2 downstream of ANXA6 in this pathway.","method":"RNA sequencing of clinical specimens, ANXA6 overexpression/knockdown, in vitro proliferation and migration assays, in vivo lymphatic metastasis assays, TRPV2 inhibitor treatment, AKT/mTOR pathway Western blot","journal":"Experimental hematology & oncology","confidence":"Medium","confidence_rationale":"Tier 2 — genetic and pharmacological epistasis with defined pathway readouts in vitro and in vivo; single lab","pmids":["37138336"],"is_preprint":false},{"year":2024,"finding":"PRRX1 was identified as a transcription factor that directly promotes ANXA6 expression in bladder cancer via ChIP-PCR and dual-luciferase assays. ANXA6 in turn promotes PKCα/EGFR complex formation (confirmed by mass spectrometry, immunofluorescence, and co-immunoprecipitation), inhibiting EGFR phosphorylation and downstream AKT and ERK1/2 signaling, thereby enhancing cisplatin sensitivity and re-sensitizing resistant cells.","method":"ChIP-PCR, dual-luciferase reporter assay for PRRX1 binding to ANXA6 promoter, mass spectrometry, immunofluorescence, co-immunoprecipitation, CCK-8, colony formation, flow cytometry (ROS, apoptosis), comet assay, xenograft model","journal":"Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods (ChIP, Co-IP, MS) confirming transcriptional regulation and downstream complex; single lab","pmids":["39528080"],"is_preprint":false},{"year":2025,"finding":"Anxa6-/- mice display rapid fasting-induced hypoglycemia due to a specific impairment in the glucose-alanine cycle: these mice cannot utilize alanine for hepatic gluconeogenesis (GNG). Fasted Anxa6-/- mice showed elevated glucagon, increased lipid oxidation, and slightly reduced expression of alanine aminotransferase 2 (Gpt2), lactate dehydrogenase (Ldha2), and the hepatic alanine transporter SNAT4, demonstrating a critical role for ANXA6 in alanine-dependent hepatic GNG.","method":"Anxa6-/- knockout mouse model, indirect calorimetry, glucose/insulin/glucagon tolerance tests, alanine challenge tests, respiratory exchange ratio measurement, biochemical enzyme expression analysis","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 — clean KO mouse with specific metabolic challenge revealing alanine-GNG defect; single lab","pmids":["40802799"],"is_preprint":false},{"year":2025,"finding":"ANXA6 overexpression in decidual macrophages promotes macrophage M1 polarization and pyroptosis through the NF-κB signaling pathway, leading to mitochondrial ROS accumulation and decreased mitochondrial membrane potential. Inhibition of NLRP3 rescued ANXA6-overexpression-induced M1 polarization and pyroptosis. ANXA6-overexpressing macrophages inhibited trophoblast function in vitro via TNF-α-mediated inhibition of the PI3K/AKT axis.","method":"ANXA6 overexpression in macrophages, ROS/mitochondrial membrane potential assays, NLRP3 inhibitor treatment, ROS inhibitor treatment, NF-κB pathway analysis, M1/M2 polarization markers, trophoblast co-culture, PI3K/AKT pathway Western blot","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological and genetic epistasis across multiple pathways; single lab","pmids":["40959268"],"is_preprint":false},{"year":2025,"finding":"MG53 binds AnxA6 at membrane injury sites, and AnxA6 overexpression abolishes MG53's cytoprotective effects in iPSCs under hypoxia/reoxygenation stress, placing AnxA6 as a negative regulator of MG53-mediated membrane repair. MG53 treatment enhanced iPSC survival and functional integration in myocardial tissue post-ischemia-reperfusion injury in vivo.","method":"Co-binding assay/interaction studies (MG53-AnxA6), AnxA6 overexpression rescue experiment, H/R injury model in iPSCs, in vivo MIRI model with MG53 treatment and iPSC transplantation","journal":"Cellular signalling","confidence":"Low","confidence_rationale":"Tier 3 — interaction inferred from functional rescue; single method for binding, single lab","pmids":["41106752"],"is_preprint":false},{"year":2025,"finding":"ANXA6 co-localizes with cGAS and acts as an inhibitor of the cGAS-STING signaling pathway by promoting K63-linked ubiquitination of cGAS, thereby reducing cGAS protein levels. ANXA6 knockdown impeded cGAS K63-ubiquitination, increased cGAS expression, activated downstream TBK1, IRF3, and type I IFN-stimulated genes (MX1, ISG15, ISG56), elevated ROS and lipid peroxides, and induced ferroptosis in cervical cancer cells.","method":"Co-localization by immunofluorescence, co-immunoprecipitation for ANXA6-cGAS interaction, ubiquitination assays (K63-linkage specificity), ANXA6 siRNA knockdown, Western blot for cGAS-STING pathway components, ROS/lipid peroxide assays, ferroptosis detection","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP plus ubiquitination assay with downstream pathway readouts; single lab","pmids":["41219880"],"is_preprint":false},{"year":2025,"finding":"In DMD skeletal muscle cells, ANXA5 and ANXA6 fail to upregulate in response to mechanical membrane stress, while control cells upregulate both proteins as part of an adaptive membrane repair mechanism. This was established by live-cell imaging of GFP-tagged annexins, Western blotting, and a shear stress-based injury assay, placing ANXA6 as part of the adaptive membrane repair response in skeletal muscle.","method":"Live-cell imaging of GFP-tagged ANXA6, shear stress-based membrane injury assay, Western blot for annexin expression, immunohistochemistry of DMD patient biopsies","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 — direct localization by live imaging tied to membrane repair function; preprint, single lab","pmids":[],"is_preprint":true},{"year":2024,"finding":"AnxA6 interacts with SNAP23 (a SNARE protein) and this interaction is required for the translocation of AnxA6 to cellular membranes and subsequent secretion of pro-inflammatory cytokines (MCP-1/CCL2, IL-8, DKK1, TSP-1, osteopontin) and small extracellular vesicles (EVs) in triple-negative breast cancer cells. AnxA6 downregulation strongly reduced EV secretion and cytokine levels; upregulation promoted EV-associated Rab7, cholesterol, and MCP-1. AnxA6 neutralizing antibodies reduced survival of AnxA6-low TNBC cells.","method":"Co-immunoprecipitation (AnxA6-SNAP23), cytokine arrays on conditioned media, EV isolation and characterization, AnxA6 siRNA knockdown and overexpression, cholesterol measurement in EVs, membrane translocation assays","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 — reciprocal Co-IP for SNAP23 interaction, combined with functional EV and cytokine secretion assays; preprint, single lab","pmids":[],"is_preprint":true}],"current_model":"ANXA6 is a Ca2+-dependent, phospholipid-binding scaffold protein that: (1) inhibits membrane fusion in a Ca2+-dependent manner and partitions into cholesterol/sphingomyelin-enriched microdomains; (2) suppresses EGFR-ERK1/2 signaling by facilitating EGFR-PKCα complex formation, an activity enhanced by SUMO1 modification at K299; (3) promotes autophagy by inhibiting the PI3K/AKT/mTOR pathway and enabling ATG9A vesicle trafficking via RAB GTPases/F-actin; (4) functions as a scaffold for the ANXA6/LRP1/TSP1 complex in cancer-associated fibroblast-derived EVs that promote pancreatic cancer aggressiveness; (5) acts as a host restriction factor for influenza A virus; (6) is required for hepatic alanine-dependent gluconeogenesis; (7) inhibits the cGAS-STING pathway by promoting K63-ubiquitination of cGAS; and (8) facilitates secretion of pro-inflammatory cytokines and EVs through interaction with SNAP23."},"narrative":{"teleology":[{"year":1988,"claim":"Determining the primary structure of ANXA6 established it as a Ca²⁺-binding annexin family member with an unusual eight-repeat architecture, providing the molecular framework for all subsequent functional studies.","evidence":"cDNA cloning and sequence analysis of human p68/ANXA6","pmids":["3258820"],"confidence":"High","gaps":["No three-dimensional structure resolved at this stage","Biological function and lipid-binding specificity not yet characterized"]},{"year":1990,"claim":"Reconstitution with defined lipid vesicles revealed that ANXA6 uniquely inhibits membrane fusion at threshold Ca²⁺ and binds selectively to anionic phospholipids, distinguishing its biophysical activity from other annexins.","evidence":"In vitro phospholipid vesicle aggregation and fusion assays with purified ANXA6","pmids":["2138016"],"confidence":"High","gaps":["Cellular consequences of fusion inhibition not demonstrated","Whether ANXA6 inhibits fusion in vivo remains untested"]},{"year":2013,"claim":"Showing that Ca²⁺ drives ANXA6 into cholesterol/sphingomyelin-enriched microdomains connected its lipid-binding activity to raft-dependent signaling and linked it mechanistically to Niemann-Pick type C pathology.","evidence":"Detergent-resistant membrane fractionation and Langmuir monolayer assays in NPC cells","pmids":["23360953"],"confidence":"Medium","gaps":["Whether ANXA6 raft partitioning is causative versus correlative in NPC disease","Structural basis for cholesterol/sphingomyelin selectivity not resolved"]},{"year":2016,"claim":"Identification of a stable ANXA6/LRP1/TSP1 complex in cancer-associated fibroblast-derived extracellular vesicles established ANXA6 as a scaffolding component of pro-tumorigenic paracrine signaling in pancreatic cancer.","evidence":"Reciprocal co-immunoprecipitation, EV uptake, ANXA6 depletion in CAFs, in vivo xenograft and metastasis assays","pmids":["27701147"],"confidence":"High","gaps":["Mechanism by which ANXA6 recruits LRP1 and TSP1 into EVs is unknown","Direct stoichiometry and structure of the ternary complex not determined"]},{"year":2018,"claim":"Anxa6-knockout mice revealed an unexpected metabolic role: despite intact insulin–Akt signaling, ANXA6 loss uncouples insulin action from suppression of hepatic gluconeogenesis, positioning ANXA6 as a fine-tuning scaffold for hepatic glucose metabolism.","evidence":"Anxa6-KO mouse model with high-fat diet, glucose/insulin tolerance tests, Akt/S6K phosphorylation, glucose production assay in AnxA6-depleted hepatocytes","pmids":["30110341"],"confidence":"High","gaps":["Direct protein target through which ANXA6 controls gluconeogenesis downstream of Akt not identified","Whether phenotype is liver-autonomous versus systemic not fully resolved"]},{"year":2020,"claim":"Two independent studies converged to show ANXA6 promotes autophagy through dual mechanisms—inhibition of PI3K/AKT/mTOR signaling and facilitation of ATG9A vesicle trafficking via RAB GTPases and F-actin—resolving how ANXA6 connects membrane dynamics to autophagosome biogenesis.","evidence":"ANXA6 knockdown/rescue with LC3 and ATG9A trafficking readouts, epistasis with PI3K inhibitor CAL101, mTOR/ERK pathway analysis","pmids":["32373608","33135350"],"confidence":"Medium","gaps":["Which RAB GTPases directly interact with ANXA6 for ATG9A sorting not defined","Structural basis for ANXA6 engagement with autophagic machinery unknown","Whether the PI3K/AKT arm and the ATG9A trafficking arm are independent or coupled is unclear"]},{"year":2021,"claim":"CRISPR knockout demonstrated that endogenous ANXA6 restricts influenza A virus replication, expanding its functional repertoire to innate antiviral defense.","evidence":"CRISPR-Cas9 ANXA6 KO in HEK293FT cells, TCID50 viral titer and growth curve assays","pmids":["34673207"],"confidence":"Medium","gaps":["Mechanism of viral restriction (virion packaging, entry, or egress) not delineated","Relevance in primary airway epithelial cells not tested","Whether ANXA6 restricts other viruses is unknown"]},{"year":2023,"claim":"Identification of K299 as a SUMO1 modification site explained how ANXA6 suppresses EGFR–ERK1/2 signaling: SUMOylated ANXA6 bridges EGFR and PKCα, and loss of this modification confers resistance to EGFR-targeted therapy, unifying the tumor-suppressive and drug-sensitizing activities of ANXA6.","evidence":"LC-MS/MS SUMOylation site mapping, K299R mutagenesis, co-IP for EGFR–PKCα complex, in vitro and xenograft assays","pmids":["37528485"],"confidence":"High","gaps":["E3 SUMO ligase responsible for K299 modification not identified","Whether SUMOylation modulates ANXA6 lipid-binding or raft partitioning is untested"]},{"year":2025,"claim":"Anxa6-KO mice challenged with alanine revealed a specific requirement for ANXA6 in the glucose–alanine cycle, pinpointing the metabolic defect to alanine-dependent hepatic gluconeogenesis and linking ANXA6 to SNAT4/Gpt2 expression.","evidence":"Anxa6-KO mouse model, alanine challenge, indirect calorimetry, transporter and enzyme expression analysis","pmids":["40802799"],"confidence":"Medium","gaps":["Whether ANXA6 directly regulates SNAT4 trafficking or Gpt2 transcription is unknown","Tissue-specific rescue experiments not reported"]},{"year":2025,"claim":"Demonstration that ANXA6 promotes K63-linked ubiquitination of cGAS to suppress the cGAS–STING–IFN axis established a new role for ANXA6 in tuning innate immune DNA sensing, with consequences for ferroptosis induction when ANXA6 is lost.","evidence":"Co-IP for ANXA6–cGAS, K63-ubiquitination assay, ANXA6 knockdown with STING pathway and ferroptosis readouts in cervical cancer cells","pmids":["41219880"],"confidence":"Medium","gaps":["E3 ubiquitin ligase recruited by ANXA6 to ubiquitinate cGAS not identified","Whether ANXA6 directly binds cGAS or acts via an intermediate is unresolved","In vivo validation of the cGAS–STING suppressive role is lacking"]},{"year":null,"claim":"The upstream signals that coordinate ANXA6's scaffolding functions across its diverse pathways—EGFR suppression, autophagy, gluconeogenesis, cGAS regulation, and membrane repair—remain mechanistically unintegrated; a unifying structural or regulatory model is absent.","evidence":"","pmids":[],"confidence":"Low","gaps":["No full-length ANXA6 structure in complex with any signaling partner exists","How SUMOylation, Ca²⁺ binding, and lipid raft partitioning are coordinately regulated is unknown","Tissue-specific isoform functions arising from alternative splicing remain largely unexplored"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[1,2]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,13,15]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[6,8,13,19]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,2,11,21]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[3,8,9]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[13,19]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[3,9,21]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[6,8,13,14,15]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[6,8,9,14]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[3,9,21]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[5,16]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[10,19]}],"complexes":["ANXA6/LRP1/TSP1 (CAF-derived EV complex)","EGFR/PKCα/ANXA6 complex"],"partners":["EGFR","PRKCA","LRP1","THBS1","SNAP23","CGAS","ATG9A","TRIM72"],"other_free_text":[]},"mechanistic_narrative":"ANXA6 is a Ca²⁺-dependent phospholipid-binding scaffold protein that organizes membrane-associated signaling platforms, regulates vesicular trafficking and autophagy, and modulates metabolic and innate immune pathways. Its eight-repeat annexin fold binds phosphatidylserine, phosphatidic acid, phosphatidylinositol, and phosphatidylethanolamine in a Ca²⁺-dependent manner and inhibits phospholipid vesicle fusion [PMID:2138016]; Ca²⁺ also drives its partitioning into cholesterol/sphingomyelin-enriched membrane microdomains [PMID:23360953]. ANXA6 suppresses EGFR–ERK1/2 signaling by facilitating PKCα–EGFR complex formation, an activity enhanced by SUMO1 modification at K299, and loss of this SUMOylation confers gefitinib resistance [PMID:37528485, PMID:39528080]. ANXA6 additionally promotes autophagy through dual inhibition of PI3K/AKT/mTOR signaling and facilitation of ATG9A vesicle trafficking via RAB GTPases/F-actin [PMID:32373608, PMID:33135350], functions as a host restriction factor for influenza A virus [PMID:34673207], is required for hepatic alanine-dependent gluconeogenesis [PMID:40802799], and inhibits the cGAS–STING pathway by promoting K63-linked ubiquitination of cGAS [PMID:41219880]."},"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,"source_track":"pubmed_title"},{"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,"source_track":"pubmed_title"},{"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":27,"is_preprint":false,"source_track":"pubmed_title"},{"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,"source_track":"pubmed_title"},{"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":25,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30110341","id":"PMC_30110341","title":"Altered hepatic glucose homeostasis in AnxA6-KO mice fed a high-fat diet.","date":"2018","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/30110341","citation_count":21,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28881357","id":"PMC_28881357","title":"A novel inhibitory anti-invasive MAb isolated using phenotypic screening highlights AnxA6 as a functionally relevant target protein in pancreatic cancer.","date":"2017","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/28881357","citation_count":21,"is_preprint":false,"source_track":"pubmed_title"},{"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":20,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36585419","id":"PMC_36585419","title":"MANF/EWSR1/ANXA6 pathway might as the bridge between hypolipidemia and major depressive disorder.","date":"2022","source":"Translational psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/36585419","citation_count":19,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26202167","id":"PMC_26202167","title":"Gene-based meta-analysis of genome-wide association study data identifies independent single-nucleotide polymorphisms in ANXA6 as being associated with systemic lupus erythematosus in Asian populations.","date":"2015","source":"Arthritis & rheumatology (Hoboken, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/26202167","citation_count":15,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37138336","id":"PMC_37138336","title":"ANXA6/TRPV2 axis promotes lymphatic metastasis in head and neck squamous cell carcinoma by inducing autophagy.","date":"2023","source":"Experimental hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/37138336","citation_count":14,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35483553","id":"PMC_35483553","title":"The mouse Anxa6/miR-9-5p/Anxa2 axis modulates TGF-β1-induced mouse hepatic stellate cell (mHSC) activation and CCl4-caused liver fibrosis.","date":"2022","source":"Toxicology letters","url":"https://pubmed.ncbi.nlm.nih.gov/35483553","citation_count":14,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37528485","id":"PMC_37528485","title":"SUMOylation of AnxA6 facilitates EGFR-PKCα complex formation to suppress epithelial cancer growth.","date":"2023","source":"Cell communication and signaling : CCS","url":"https://pubmed.ncbi.nlm.nih.gov/37528485","citation_count":13,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33425920","id":"PMC_33425920","title":"Enterohemorrhagic Escherichia coli Effector Protein EspF Interacts With Host Protein ANXA6 and Triggers Myosin Light Chain Kinase (MLCK)-Dependent Tight Junction Dysregulation.","date":"2020","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/33425920","citation_count":12,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36361965","id":"PMC_36361965","title":"Apigenin Modulates AnxA6- and TNAP-Mediated Osteoblast Mineralization.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36361965","citation_count":10,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33582672","id":"PMC_33582672","title":"The Impact of ANxA6 Gene Polymorphism on the Efficacy of Methotrexate Treatment in Psoriasis Patients.","date":"2021","source":"Dermatology (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/33582672","citation_count":10,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36160712","id":"PMC_36160712","title":"EZH2 Regulates ANXA6 Expression via H3K27me3 and Is Involved in Angiotensin II-Induced Vascular Smooth Muscle Cell Senescence.","date":"2022","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/36160712","citation_count":9,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34673207","id":"PMC_34673207","title":"CRISPR-Cas9 mediated knockout of AnxA6 gene enhances influenza A virus replication in low-permissive HEK293FT cell line.","date":"2021","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/34673207","citation_count":8,"is_preprint":false,"source_track":"pubmed_title"},{"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,"source_track":"pubmed_title"},{"pmid":"39577524","id":"PMC_39577524","title":"ANXA6 expression as a potential indicator of tumor diagnosis, metastasis and immunity in nasopharyngeal carcinoma.","date":"2024","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/39577524","citation_count":5,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37311953","id":"PMC_37311953","title":"Comprehensive analysis the diagnosis, malignant progression and immune infiltrate of ANXA6 in prostate cancer.","date":"2023","source":"Genes & genomics","url":"https://pubmed.ncbi.nlm.nih.gov/37311953","citation_count":3,"is_preprint":false,"source_track":"pubmed_title"},{"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":3,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"40802799","id":"PMC_40802799","title":"Fasting-Induced Hepatic Gluconeogenesis Is Compromised In Anxa6-/- Mice.","date":"2025","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/40802799","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"40989757","id":"PMC_40989757","title":"Identification of Core SASP-Related Genes TGFBI and ANXA6 for Diagnosing Ulcerative Colitis-Related Colorectal Cancer Through Machine Learning.","date":"2025","source":"Journal of inflammation research","url":"https://pubmed.ncbi.nlm.nih.gov/40989757","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36998449","id":"PMC_36998449","title":"The polymorphisms of ANXA6 influence head and neck cancer susceptibility in the Chinese Han population.","date":"2023","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/36998449","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"40959268","id":"PMC_40959268","title":"ANXA6 Overexpression Causes Abnormal Decidual Macrophage-Trophoblast Crosstalk in Recurrent Spontaneous Abortion.","date":"2025","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40959268","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"41876560","id":"PMC_41876560","title":"Multi-omics integration reveals ANXA6-high γδ T cell-endothelial communication as a potential link between periodontitis and MASLD.","date":"2026","source":"NPJ systems biology and applications","url":"https://pubmed.ncbi.nlm.nih.gov/41876560","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"41106752","id":"PMC_41106752","title":"Mitsugumin 53 improves myocardial ischemia-reperfusion injury by promoting iPSCs survival through regulating AnxA6 axis.","date":"2025","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/41106752","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"41219880","id":"PMC_41219880","title":"Mechanism of interaction between ANXA6 and cGAS to regulate STING signaling pathway and promote ferroptosis in cervical cancer.","date":"2025","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/41219880","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"40374359","id":"PMC_40374359","title":"[Ursolic acid down-regulates ANXA6 and inhibits breast cancer cell growth and metastasis].","date":"2025","source":"Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine]","url":"https://pubmed.ncbi.nlm.nih.gov/40374359","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38188996","id":"PMC_38188996","title":"Retracted: EZH2 Regulates ANXA6 Expression via H3K27me3 and Is Involved in Angiotensin II-Induced Vascular Smooth Muscle Cell Senescence.","date":"2023","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/38188996","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"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,"source_track":"pubmed_title"},{"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,"source_track":"pubmed_title"},{"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 splicing","date":"2025-06-19","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.15.659434","citation_count":0,"is_preprint":true,"source_track":"pubmed_title"},{"pmid":null,"id":"bio_10.1101_2025.11.01.685128","title":"Contribution of Mesenchymal-like and Epithelial Cellular Subsets to Chemotherapy Resistance in Triple-Negative Breast Cancer","date":"2025-11-03","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.01.685128","citation_count":0,"is_preprint":true,"source_track":"pubmed_title"},{"pmid":null,"id":"bio_10.1101_2024.10.22.619710","title":"Annexin A6 modulates the secretion of pro-inflammatory cytokines and exosomes via interaction with SNAP23 in triple negative breast cancer cells","date":"2024-10-25","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.22.619710","citation_count":0,"is_preprint":true,"source_track":"pubmed_title"},{"pmid":"16169070","id":"PMC_16169070","title":"A human protein-protein interaction network: a resource for annotating the proteome.","date":"2005","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/16169070","citation_count":1704,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12477932","id":"PMC_12477932","title":"Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/12477932","citation_count":1479,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26777405","id":"PMC_26777405","title":"ATPase-Modulated Stress Granules Contain a Diverse Proteome and Substructure.","date":"2016","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/26777405","citation_count":1233,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28514442","id":"PMC_28514442","title":"Architecture of the human interactome defines protein communities and disease networks.","date":"2017","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/28514442","citation_count":1085,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"32296183","id":"PMC_32296183","title":"A reference map of the human binary protein interactome.","date":"2020","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/32296183","citation_count":849,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"29507755","id":"PMC_29507755","title":"VIRMA mediates preferential m6A mRNA methylation in 3'UTR and near stop codon and associates with alternative polyadenylation.","date":"2018","source":"Cell discovery","url":"https://pubmed.ncbi.nlm.nih.gov/29507755","citation_count":829,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"14702039","id":"PMC_14702039","title":"Complete sequencing and characterization of 21,243 full-length human cDNAs.","date":"2003","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/14702039","citation_count":754,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"33961781","id":"PMC_33961781","title":"Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.","date":"2021","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/33961781","citation_count":705,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21873635","id":"PMC_21873635","title":"Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium.","date":"2011","source":"Briefings in bioinformatics","url":"https://pubmed.ncbi.nlm.nih.gov/21873635","citation_count":656,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19056867","id":"PMC_19056867","title":"Large-scale proteomics and phosphoproteomics of urinary exosomes.","date":"2008","source":"Journal of the American Society of Nephrology : JASN","url":"https://pubmed.ncbi.nlm.nih.gov/19056867","citation_count":607,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"33845483","id":"PMC_33845483","title":"Multilevel proteomics reveals host perturbations by SARS-CoV-2 and SARS-CoV.","date":"2021","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/33845483","citation_count":532,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21423176","id":"PMC_21423176","title":"Analysis of the myosin-II-responsive focal adhesion proteome reveals a role for β-Pix in negative regulation of focal adhesion maturation.","date":"2011","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/21423176","citation_count":490,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15489334","id":"PMC_15489334","title":"The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC).","date":"2004","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/15489334","citation_count":438,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"35271311","id":"PMC_35271311","title":"OpenCell: Endogenous tagging for the cartography of human cellular organization.","date":"2022","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/35271311","citation_count":432,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26871637","id":"PMC_26871637","title":"Widespread Expansion of Protein Interaction Capabilities by Alternative Splicing.","date":"2016","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/26871637","citation_count":423,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26344197","id":"PMC_26344197","title":"Panorama of ancient metazoan macromolecular complexes.","date":"2015","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/26344197","citation_count":407,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"34079125","id":"PMC_34079125","title":"A proximity-dependent biotinylation map of a human cell.","date":"2021","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/34079125","citation_count":339,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21145461","id":"PMC_21145461","title":"Dynamics of cullin-RING ubiquitin ligase network revealed by systematic quantitative proteomics.","date":"2010","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/21145461","citation_count":318,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28611215","id":"PMC_28611215","title":"Genome-wide CRISPR screen identifies HNRNPL as a prostate cancer dependency regulating RNA splicing.","date":"2017","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/28611215","citation_count":282,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"2138016","id":"PMC_2138016","title":"Characterization of Ca2(+)-dependent phospholipid binding, vesicle aggregation and membrane fusion by annexins.","date":"1990","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/2138016","citation_count":276,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"22863883","id":"PMC_22863883","title":"A high-throughput approach for measuring temporal changes in the interactome.","date":"2012","source":"Nature methods","url":"https://pubmed.ncbi.nlm.nih.gov/22863883","citation_count":273,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"35063084","id":"PMC_35063084","title":"Tau interactome maps synaptic and mitochondrial processes associated with neurodegeneration.","date":"2022","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/35063084","citation_count":256,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"20458337","id":"PMC_20458337","title":"MHC class II-associated proteins in B-cell exosomes and potential functional implications for exosome biogenesis.","date":"2010","source":"Immunology and cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/20458337","citation_count":221,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"25037231","id":"PMC_25037231","title":"Extracellular matrix signatures of human primary metastatic colon cancers and their metastases to liver.","date":"2014","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/25037231","citation_count":203,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"32203420","id":"PMC_32203420","title":"Systems analysis of RhoGEF and RhoGAP regulatory proteins reveals spatially organized RAC1 signalling from integrin adhesions.","date":"2020","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/32203420","citation_count":194,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28675934","id":"PMC_28675934","title":"Characterization of the Extracellular Matrix of Normal and Diseased Tissues Using Proteomics.","date":"2017","source":"Journal of proteome research","url":"https://pubmed.ncbi.nlm.nih.gov/28675934","citation_count":185,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"27701147","id":"PMC_27701147","title":"Cancer-associated fibroblast-derived annexin A6+ extracellular vesicles support pancreatic cancer aggressiveness.","date":"2016","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/27701147","citation_count":182,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"3258820","id":"PMC_3258820","title":"Primary structure of the human, membrane-associated Ca2+-binding protein p68 a novel member of a protein family.","date":"1988","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/3258820","citation_count":161,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"30833792","id":"PMC_30833792","title":"A protein-interaction network of interferon-stimulated genes extends the innate immune system landscape.","date":"2019","source":"Nature immunology","url":"https://pubmed.ncbi.nlm.nih.gov/30833792","citation_count":159,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"17081065","id":"PMC_17081065","title":"Proteomic and bioinformatic characterization of the biogenesis and function of melanosomes.","date":"2006","source":"Journal of proteome research","url":"https://pubmed.ncbi.nlm.nih.gov/17081065","citation_count":150,"is_preprint":false,"source_track":"gene2pubmed"}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":23175,"output_tokens":4356,"usd":0.067433},"stage2":{"model":"claude-opus-4-6","input_tokens":7799,"output_tokens":3037,"usd":0.17238},"total_usd":0.552823,"stage1_batch_id":"msgbatch_01TqNM6p6FsoMBT2jzQTwKBr","stage2_batch_id":"msgbatch_01Ya473eco8sTHbvRymQaU8o","note":"batch pricing = 50% of standard","round2_stage1":{"model":"claude-sonnet-4-6","input_tokens":35263,"output_tokens":8194,"usd":0.11435},"round2_rules_fired":"R3","round2_stage2":{"model":"claude-opus-4-6","input_tokens":10363,"output_tokens":3225,"usd":0.19866}},"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, as demonstrated by siRNA-mediated knockdown activating the PI3K/AKT/mTOR pathway and reducing autophagy; the PI3K inhibitor CAL101 reversed siANXA6-reduced autophagy and also increased ANXA6 expression in a negative feedback loop.\",\n      \"method\": \"siRNA knockdown, pharmacological inhibition (CAL101), Western blot for pathway activity, LC3 siRNA epistasis\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KD with defined cellular phenotype and pharmacological epistasis, single lab\",\n      \"pmids\": [\"32373608\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ANXA6 regulates autophagosome formation by enabling ATG9A-positive vesicle trafficking from recycling endosomes to autophagosomes through RAB GTPases and F-actin; ANXA6 also suppresses mTOR activity by inhibiting PI3K-AKT and ERK signaling pathways.\",\n      \"method\": \"siRNA knockdown, ANXA6 restoration, GO analysis of ANXA6 interactors (RAB GTPases), ATG9A trafficking assays, signaling pathway Western blot\",\n      \"journal\": \"Clinical and translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KD/rescue with specific vesicle trafficking readout and pathway analysis, single lab\",\n      \"pmids\": [\"33135350\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ANXA6 is SUMOylated at lysine K299 by SUMO1; SUMOylated ANXA6 preferentially binds EGFR upon EGF stimulation and facilitates EGFR-PKCα complex formation, leading to decreased phosphorylation of EGFR and ERK1/2, and reduced cyclin D1 expression. The SUMO-site mutant AnxA6K299R fails to suppress EGFR-ERK1/2 phosphorylation.\",\n      \"method\": \"Co-transfection with SUMO1/UBC9/AnxA6 plasmids, immunoprecipitation, LC-MS/MS identification of SUMOylation sites, site-directed mutagenesis (K299R), Western blot, lentiviral shRNA KD and OE, xenograft tumor model\",\n      \"journal\": \"Cell communication and signaling : CCS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — SUMOylation site identified by MS and validated by mutagenesis, Co-IP for complex, functional phenotype in vitro and in vivo\",\n      \"pmids\": [\"37528485\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Transcription factor PRRX1 drives ANXA6 expression; ANXA6 promotes PKCα/EGFR complex formation, inhibiting EGFR phosphorylation and downstream AKT and ERK1/2, thereby enhancing cisplatin sensitivity in bladder cancer cells.\",\n      \"method\": \"Mass spectrometry, co-immunoprecipitation, immunofluorescence, ChIP-PCR, dual-luciferase reporter assay, CCK-8, xenograft model\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP and MS for complex, ChIP for transcriptional regulation, in vivo validation; single lab\",\n      \"pmids\": [\"39528080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"The bacterial effector EspF interacts with host ANXA6 through EspF's C-terminal domain; co-expression of EspF-ANXA6 activates myosin light chain kinase (MLCK), induces phosphorylation of myosin light chain (MLC) and PKCα, downregulates calmodulin, and decreases tight junction proteins ZO-1 and occludin, disrupting intestinal barrier integrity.\",\n      \"method\": \"FRET, co-immunoprecipitation, Western blot for TJ proteins and phosphorylation status\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — FRET and Co-IP for direct interaction, multiple downstream readouts; single lab\",\n      \"pmids\": [\"33425920\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ANXA6 acts as a host restriction factor for influenza A virus; CRISPR-Cas9 knockout of ANXA6 in HEK293FT cells significantly increased influenza A virus replication titers, establishing ANXA6 as an inhibitor of influenza virion packaging.\",\n      \"method\": \"CRISPR-Cas9 knockout, TCID assay, flow cytometry, viral growth curve experiments\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with defined viral replication phenotype, orthogonal quantification methods; single lab\",\n      \"pmids\": [\"34673207\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"AnxA6 contributes to hepatic glucose metabolism; AnxA6-KO mice on high-fat diet show increased insulin-stimulated Akt and S6K phosphorylation in liver, but fail to downregulate hepatic gluconeogenesis despite normal insulin signaling, and have increased glycogen storage. AnxA6-depleted HuH7 hepatocytes are unable to reduce glucose production upon insulin exposure.\",\n      \"method\": \"AnxA6 knockout mouse model, HFD feeding, OGTT/ITT, Western blot for Akt/S6K phosphorylation, hepatocyte glucose production assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with multiple metabolic readouts and in vitro validation; single lab, moderate evidence\",\n      \"pmids\": [\"30110341\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Anxa6-knockout mice display rapid hypoglycemia during fasting due to impaired alanine-dependent hepatic gluconeogenesis; Anxa6-/- mice cannot utilize alanine for glucose production, correlating with reduced expression of alanine aminotransferase 2 (Gpt2), lactate dehydrogenase (Ldha2), and hepatic alanine transporter SNAT4.\",\n      \"method\": \"Anxa6 knockout mouse model, indirect calorimetry, glucose/insulin/pyruvate/alanine tolerance tests, enzyme expression analysis\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with multiple metabolic phenotyping methods; single lab\",\n      \"pmids\": [\"40802799\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"AnxA6 associates with lipid microdomains (detergent-resistant membranes) in a calcium-dependent manner; in Niemann-Pick type C disease cells, AnxA6 translocates to cholesterol/sphingomyelin-enriched DRMs upon Ca2+ rise. In vitro Langmuir monolayer experiments showed Ca2+ is the primary regulator of AnxA6 interaction with DPPC and sphingomyelin monolayers.\",\n      \"method\": \"DRM isolation by sucrose gradient fractionation, Langmuir monolayer assay, calcium titration\",\n      \"journal\": \"Molecular bioSystems\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro reconstitution with monolayers plus cell fractionation; single lab\",\n      \"pmids\": [\"23360953\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"EZH2 represses ANXA6 expression by increasing H3K27me3 modification at the ANXA6 promoter; loss of EZH2 allows ANXA6 upregulation, promoting ROS production, G1 arrest, and vascular smooth muscle cell senescence. Overexpression of EZH2 reversed these effects, which were partially rescued by simultaneous ANXA6 overexpression.\",\n      \"method\": \"ChIP for H3K27me3 at ANXA6 promoter, EZH2 overexpression/knockdown, ANXA6 overexpression/knockdown, SA-β-gal staining, flow cytometry, DHE staining for ROS, mouse AAA model\",\n      \"journal\": \"Oxidative medicine and cellular longevity\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP demonstrates epigenetic mechanism, epistasis established by dual OE; note: paper was later retracted (PMID:38188996)\",\n      \"pmids\": [\"36160712\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ANXA6 co-localizes with cGAS and promotes ubiquitination of cGAS (specifically inhibiting K63-linked ubiquitination), thereby suppressing the cGAS-STING innate immune signaling pathway. ANXA6 knockdown relieves cGAS ubiquitination, activates cGAS-STING, elevates phospho-TBK1, IRF3, and ISGs, increases ROS and lipid peroxides, and induces ferroptosis in cervical cancer cells.\",\n      \"method\": \"Co-localization assays, co-immunoprecipitation, ubiquitination assays, ANXA6 knockdown, Western blot for TBK1/IRF3/ISG pathway\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP for interaction, ubiquitination assay for mechanism, KD with defined downstream signaling readouts; single lab\",\n      \"pmids\": [\"41219880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ANXA6 promotes lymphatic metastasis in head and neck squamous cell carcinoma by inducing autophagy through inhibition of the AKT/mTOR signaling pathway; ANXA6 expression positively correlates with TRPV2 expression, and TRPV2 inhibition reverses ANXA6-induced autophagy and metastasis.\",\n      \"method\": \"In vitro proliferation/migration assays, in vivo lymph node metastasis model, AKT/mTOR pathway Western blot, TRPV2 inhibition, ANXA6 overexpression/knockdown\",\n      \"journal\": \"Experimental hematology & oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vitro and in vivo KD/OE with pathway readouts and epistasis via TRPV2 inhibition; single lab\",\n      \"pmids\": [\"37138336\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Apigenin disrupts intracellular distribution of AnxA6, blocking its aggregation and inhibiting its attachment to membranes in matrix vesicles, thereby modulating AnxA6- and TNAP-mediated osteoblast mineralization.\",\n      \"method\": \"Fluorescence microscopy (FM) of cells, TEM-gold immunolabeling of vesicles, AR-S/CPC mineralization assay, TNAP ELISA, TEM-EDX mineral composition\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — pharmacological perturbation with localization readout, no direct mutagenesis of AnxA6\",\n      \"pmids\": [\"36361965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MG53 (mitsugumin 53) binds AnxA6 and stabilizes damaged membranes; AnxA6 overexpression abolishes MG53's cytoprotective effects on iPSCs under hypoxia/reoxygenation injury, positioning AnxA6 as a negative regulator downstream of MG53 in membrane repair.\",\n      \"method\": \"Co-immunoprecipitation (MG53-AnxA6 interaction), AnxA6 overexpression rescue experiments, in vivo iPSC survival in MIRI mouse model, apoptosis assays\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single Co-IP with limited mechanistic follow-up; epistasis by OE only\",\n      \"pmids\": [\"41106752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Control skeletal muscle cells upregulate ANXA5 and ANXA6 upon mechanical stress as part of an adaptive membrane repair response; DMD cells fail to induce this ANXA6 upregulation after membrane injury, indicating ANXA6 participates in membrane resealing.\",\n      \"method\": \"Shear stress-based injury assay, live-cell imaging of GFP-tagged annexins, Western blotting, immunohistochemistry of DMD patient biopsies\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — observational upregulation in stress response, no direct gain/loss of ANXA6 function tested; preprint\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"AnxA6 facilitates secretion of pro-inflammatory cytokines (MCP-1/CCL2, IL-8) and small extracellular vesicles in TNBC cells through interaction with SNAP23; AnxA6 downregulation reduces EV-associated Rab7, cholesterol, and MCP-1. AnxA6 translocation to cellular membranes is required for SNAP23 interaction and EV/cytokine secretion.\",\n      \"method\": \"Cytokine arrays, EV isolation and characterization, co-immunoprecipitation (AnxA6-SNAP23), AnxA6 KD/OE, cholesterol measurement in EVs\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single Co-IP for SNAP23 interaction; preprint, single lab\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"A function-blocking monoclonal antibody (MAb 9E1) targeting AnxA6 on the cell surface significantly decreases invasion of pancreatic, lung squamous, and breast cancer cells in vitro; siRNA silencing of AnxA6 markedly reduces invasive capacity of pancreatic and lung squamous cancer cells, establishing AnxA6 as a functionally relevant pro-invasive target.\",\n      \"method\": \"Immunoprecipitation/LC-MS/MS for target identification, anti-AnxA6 MAb functional blocking, siRNA silencing, invasion assays\",\n      \"journal\": \"British journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — MS-confirmed target, antibody blocking and siRNA KD with defined invasion phenotype; single lab\",\n      \"pmids\": [\"28881357\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ANXA6 is a Ca2+-dependent membrane- and phospholipid-binding scaffold protein that: (1) suppresses EGFR-ERK1/2 signaling by facilitating EGFR-PKCα complex formation (promoted by SUMO1 modification at K299); (2) inhibits PI3K-AKT/mTOR signaling to promote autophagy and modulate vesicle trafficking including ATG9A-positive endosomal trafficking required for autophagosome formation; (3) promotes secretion of pro-inflammatory cytokines and extracellular vesicles via interaction with SNAP23; (4) restricts influenza A virus replication; (5) interacts with cGAS to promote its ubiquitination and suppress cGAS-STING innate immune signaling; (6) contributes to hepatic gluconeogenesis, particularly alanine-dependent glucose production during fasting; and (7) participates in membrane repair responses in skeletal muscle.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\nI'll classify each paper before extracting discoveries.\n\n**Classification:**\n\n- Papers [1-3] (retrieved): KEEP - ANXA6 mechanistic studies\n- Paper [4] (retrieved): KEEP - chicken ANXA6 ortholog, mechanistic (miR-24-3p/ANXA6 axis)\n- Paper [5] (retrieved): EXCLUDE - GWAS/association study, no mechanism\n- Paper [6] (retrieved): KEEP - AnxA6-KO mouse metabolic study\n- Paper [7] (retrieved): KEEP - AnxA6 invasion/cancer mechanism\n- Paper [8] (retrieved): KEEP - review, but grounded in corpus mechanisms\n- Paper [9] (retrieved): EXCLUDE - bioinformatics/biomarker, no direct mechanism\n- Paper [10] (retrieved): EXCLUDE - GWAS association\n- Paper [11] (retrieved): KEEP - ANXA6/TRPV2/autophagy mechanism\n- Paper [12] (retrieved): EXCLUDE - describes mouse lncRNA ANXA2P2 labeled \"mouse Anxa6\" - this is an alt-locus product (lncRNA), case (B)\n- Paper [13] (retrieved): KEEP - SUMOylation of AnxA6 mechanism\n- Paper [14] (retrieved): KEEP - ANXA6/EspF interaction mechanism\n- Paper [15] (retrieved): KEEP - AnxA6/TNAP mineralization mechanism\n- Paper [16] (retrieved): EXCLUDE - pharmacogenomics/SNP association\n- Paper [17] (retrieved): KEEP - EZH2/H3K27me3/ANXA6 mechanism (NOTE: paper [31] retracts this - EXCLUDE both [17] and [31])\n- Paper [18] (retrieved): KEEP - ANXA6 restriction of influenza\n- Paper [19] (retrieved): KEEP - AnxA6 lipid microdomain interaction\n- Paper [20] (retrieved): EXCLUDE - expression/biomarker study\n- Paper [21] (retrieved): EXCLUDE - expression/bioinformatics\n- Paper [22] (retrieved): KEEP - ANXA6 promotes cisplatin sensitivity via PKCα/EGFR\n- Paper [23] (retrieved): KEEP - AnxA6/gluconeogenesis KO study\n- Paper [24] (retrieved): EXCLUDE - machine learning/biomarker\n- Paper [25] (retrieved): EXCLUDE - SNP association\n- Paper [26] (retrieved): KEEP - ANXA6 in macrophage pyroptosis/RSA\n- Paper [27] (retrieved): EXCLUDE - multi-omics association\n- Paper [28] (retrieved): KEEP - MG53/AnxA6 membrane repair\n- Paper [29] (retrieved): KEEP - ANXA6/cGAS/STING mechanism\n- Paper [30] (retrieved): KEEP - UA/ANXA6/breast cancer mechanism\n- Paper [31] (retrieved): EXCLUDE - retraction notice\n- Paper [32] (preprint): KEEP - ANXA5/ANXA6 membrane repair in DMD\n- Paper [33] (preprint): KEEP - same as [23], preprint version\n- Paper [34] (preprint): EXCLUDE - alt-locus product (nt-RNA isoform), case (B)\n- Paper [35] (preprint): KEEP - AnxA6 as marker, minimal mechanism\n- Paper [36] (preprint): KEEP - AnxA6/SNAP23 secretion mechanism\n\n**gene2pubmed papers:**\n- [g1] PMID:16169070: KEEP - Y2H interactome (ANXA6 interactions identified)\n- [g2] PMID:12477932: EXCLUDE - cDNA sequencing resource\n- [g3] PMID:26777405: EXCLUDE - stress granule proteome (ANXA6 incidental)\n- [g4] PMID:28514442: EXCLUDE - general interactome resource\n- [g5] PMID:32296183: EXCLUDE - general interactome resource\n- [g6] PMID:29507755: EXCLUDE - m6A methylation, not ANXA6\n- [g7] PMID:14702039: EXCLUDE - cDNA resource\n- [g8] PMID:33961781: EXCLUDE - general interactome\n- [g9] PMID:21873635: EXCLUDE - GO annotation methods\n- [g10] PMID:19056867: EXCLUDE - urinary exosome proteome (ANXA6 incidental)\n- [g11] PMID:33845483: EXCLUDE - SARS-CoV-2 proteomics (ANXA6 incidental)\n- [g12] PMID:21423176: EXCLUDE - focal adhesion proteome (ANXA6 incidental)\n- [g13] PMID:15489334: EXCLUDE - cDNA resource\n- [g14] PMID:35271311: EXCLUDE - general cell map resource\n- [g15] PMID:26871637: EXCLUDE - alternative splicing resource\n- [g16] PMID:26344197: EXCLUDE - general complex resource\n- [g17] PMID:34079125: EXCLUDE - general BioID map\n- [g18] PMID:21145461: EXCLUDE - CRL network\n- [g19] PMID:28611215: EXCLUDE - HNRNPL/prostate cancer\n- [g20] PMID:2138016: KEEP - foundational annexin biochemistry including p68/ANXA6\n- [g21] PMID:22863882: EXCLUDE - general interactome method\n- [g22] PMID:35063084: EXCLUDE - Tau interactome\n- [g23] PMID:20458337: EXCLUDE - B-cell exosome proteome\n- [g24] PMID:25037231: EXCLUDE - ECM proteomics\n- [g25] PMID:32203420: EXCLUDE - RhoGEF/RhoGAP\n- [g26] PMID:28675934: EXCLUDE - ECM proteomics\n- [g27] PMID:27701147: KEEP - ANXA6/LRP1/TSP1 complex in CAF EVs\n- [g28] PMID:3258820: KEEP - primary structure of p68/ANXA6\n- [g29] PMID:30833792: EXCLUDE - ISG network (ANXA6 incidental)\n- [g30] PMID:17081065: EXCLUDE - melanosome proteome\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1988,\n      \"finding\": \"The primary amino acid sequence of human p68 (later named ANXA6) was determined from cDNA clones, revealing it is a membrane-associated Ca2+-binding protein with an internally repetitive structure comprising eight repeats, each containing a conserved sequence motif. It was identified as a member of the annexin family, related to p36, lipocortin I, and protein II, and the cDNA data suggested alternatively spliced mRNAs could encode different p68 protein species.\",\n      \"method\": \"cDNA cloning, sequencing, and sequence analysis\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — original structural characterization by cDNA sequencing with functional inference; foundational paper (161 citations)\",\n      \"pmids\": [\"3258820\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1990,\n      \"finding\": \"ANXA6 (referred to as p68) was shown to inhibit phospholipid vesicle membrane fusion at threshold Ca2+ concentrations, distinguishing it functionally from other annexins. It binds Ca2+-dependently to phosphatidic acid, phosphatidylserine, phosphatidylinositol, and phosphatidylethanolamine vesicles, but not phosphatidylcholine vesicles. These quantitative and qualitative differences among annexins suggest distinct intracellular functions.\",\n      \"method\": \"In vitro phospholipid vesicle binding assay, vesicle aggregation and fusion assays with defined lipid compositions and Ca2+ concentrations\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct in vitro reconstitution with quantitative lipid binding and fusion assays; highly cited (276 citations)\",\n      \"pmids\": [\"2138016\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"In Niemann-Pick type C (NPC) disease cells, AnxA6 translocates to lipid microdomains (detergent-resistant membranes, DRMs) enriched in cholesterol and sphingomyelin upon a rise in Ca2+ concentration. In vitro Langmuir monolayer experiments showed that Ca2+ is the primary factor regulating AnxA6 interaction with neutral lipid monolayers (DPPC and sphingomyelin), indicating Ca2+-driven partitioning of AnxA6 into cholesterol/sphingomyelin-enriched microdomains contributes to NPC pathology.\",\n      \"method\": \"Lipid microdomain (DRM) isolation and fractionation, Langmuir monolayer in vitro lipid-protein interaction assays, calcium titration\",\n      \"journal\": \"Molecular bioSystems\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — cell fractionation combined with Tier 1 in vitro reconstitution; single lab\",\n      \"pmids\": [\"23360953\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Cancer-associated fibroblasts (CAFs) secrete ANXA6-containing extracellular vesicles (EVs) that carry an ANXA6/LRP1/TSP1 complex. Formation of this complex is restricted to CAFs under physiopathologic conditions. Tumor cell uptake of CAF-derived ANXA6+ EVs promotes pancreatic ductal adenocarcinoma (PDA) cell survival and migration. Depletion of ANXA6 in CAFs impaired complex formation and reduced PDA aggressiveness and metastasis in vivo, while injection of ANXA6+ EVs enhanced tumorigenesis.\",\n      \"method\": \"Proteomic stromal signature analysis, co-immunoprecipitation to identify the ANXA6/LRP1/TSP1 complex, ANXA6 depletion in CAFs, in vivo xenograft and metastasis assays, EV uptake experiments\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, in vivo rescue experiments, multiple orthogonal methods; highly cited (182 citations)\",\n      \"pmids\": [\"27701147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"AnxA6 was identified as the target antigen of the inhibitory monoclonal antibody MAb 9E1 using immunoprecipitation/LC-MS/MS. Silencing of AnxA6 markedly reduced the invasive capacity of pancreatic and lung squamous cancer cells in vitro, identifying AnxA6 as a functionally relevant pro-invasive target at the cancer cell surface.\",\n      \"method\": \"Immunoprecipitation/LC-MS/MS target identification, AnxA6 siRNA knockdown, invasion assays in vitro\",\n      \"journal\": \"British journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — MS-based target identification combined with functional KD; single lab\",\n      \"pmids\": [\"28881357\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"AnxA6-knockout (KO) mice fed a high-fat diet gained less weight and showed reduced adiposity compared to wild-type controls. AnxA6-KO mice displayed increased insulin-inducible phosphorylation of Akt and S6K in liver, but failed to downregulate hepatic gluconeogenesis despite normal insulin signaling. AnxA6-depleted HuH7 hepatocytes were unable to reduce glucose production upon insulin exposure, and KO animals showed increased hepatic glycogen storage, demonstrating that AnxA6 contributes to fine-tuning of hepatic glucose metabolism.\",\n      \"method\": \"AnxA6-KO mouse model, high-fat diet feeding, oral glucose tolerance tests (OGTT), insulin tolerance tests (ITT), Western blot for Akt/S6K phosphorylation, glucose production assay in AnxA6-depleted hepatocytes\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean KO mouse with defined metabolic phenotypes, confirmed in vitro in hepatocytes with multiple readouts\",\n      \"pmids\": [\"30110341\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ANXA6 promotes autophagy by inhibiting the PI3K/AKT/mTOR signaling pathway in nasopharyngeal carcinoma cells, thereby contributing to radioresistance. ANXA6 siRNA suppressed autophagy through activation (de-repression) of PI3K/AKT/mTOR, and the PI3K inhibitor CAL101 reversed siANXA6-reduced autophagy. A negative feedback loop was identified in which suppression of PI3K/AKT/mTOR by CAL101 increased ANXA6 expression.\",\n      \"method\": \"ANXA6 siRNA knockdown, LC3 siRNA knockdown, PI3K/AKT/mTOR inhibitor (CAL101) treatment, autophagy assays, Western blot for pathway components\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological and genetic epistasis with defined pathway readouts; single lab\",\n      \"pmids\": [\"32373608\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EHEC effector protein EspF physically interacts with host protein ANXA6 through its C-terminal domain, as verified by FRET and co-immunoprecipitation. Co-expression of EspF-ANXA6 activated myosin light chain kinase (MLCK), induced phosphorylation of myosin light chain (MLC) and PKCα, downregulated Calmodulin, and decreased levels and disrupted distribution of tight junction proteins ZO-1 and occludin, revealing a mechanism by which bacterial infection perturbs epithelial barrier function through ANXA6.\",\n      \"method\": \"FRET, co-immunoprecipitation, domain mapping, Western blot for MLCK/MLC/PKCα phosphorylation and tight junction proteins, confocal microscopy for ZO-1 distribution\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — FRET plus co-IP for interaction, combined with downstream pathway readouts; single lab\",\n      \"pmids\": [\"33425920\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ANXA6 enables appropriate ATG9A+ vesicle trafficking from endosomes to autophagosomes through RAB GTPases and/or F-actin, thereby facilitating autophagosome formation. ANXA6 knockdown attenuated starvation-induced autophagy, while restoration of expression enhanced it. ANXA6 also suppresses mTOR activity through inhibition of PI3K-AKT and ERK signaling pathways, providing dual control of autophagy.\",\n      \"method\": \"ANXA6 siRNA knockdown, ANXA6 rescue overexpression, GO analysis of ANXA6 interaction targets, ATG9A vesicle trafficking assays, mTOR/PI3K-AKT/ERK signaling Western blot, LC3 reporter assays\",\n      \"journal\": \"Clinical and translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KD/rescue with multiple readouts; GO-based interactome inference supports mechanism; single lab\",\n      \"pmids\": [\"33135350\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ANXA6-containing exosomes (ANXA6-exo) derived from breast cancer stem cells promote paclitaxel resistance and cancer progression in recipient cells via autophagy induction in a YAP1-dependent manner. ANXA6-exo upregulated YAP1 to dysregulate the Hippo pathway, and silencing YAP1 abrogated the resistance effects. Autophagy inhibition with 3-methyladenine blocked ANXA6-exo-mediated PTX resistance.\",\n      \"method\": \"Exosome isolation and transfer experiments, ANXA6 and YAP1 siRNA knockdown, autophagy inhibitor (3-methyladenine), paclitaxel resistance assays, stem cell enrichment assays\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — exosome transfer with genetic and pharmacological epistasis; single lab\",\n      \"pmids\": [\"34676207\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CRISPR-Cas9 knockout of the ANXA6 gene in HEK293FT cells significantly increased influenza A virus replication and accumulation, demonstrating that endogenous ANXA6 functions as a host restriction factor that counteracts influenza virion packaging.\",\n      \"method\": \"CRISPR-Cas9 gene knockout, TCID50 viral titer assay, flow cytometry, viral growth curve experiments in ANXA6-/- versus parental cells\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean CRISPR KO with defined viral replication phenotype; single lab\",\n      \"pmids\": [\"34673207\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In osteoblasts, AnxA6 plays a role in matrix vesicle-mediated mineralization. Apigenin treatment disrupted intracellular distribution of AnxA6 by blocking AnxA6 aggregation and TNAP attachment to membranes, and modulated TNAP enzymatic activity, affecting mineral composition and structure. AnxA6 distribution was examined by fluorescence microscopy and TEM-gold in cells and vesicles.\",\n      \"method\": \"Fluorescence microscopy (FM), TEM-gold labeling of vesicles, ELISA for TNAP activity, AR-S/CPC mineralization assay, apigenin pharmacological treatment in hFOB1.19, Saos-2, and HCASMC cell lines\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization by TEM-gold and FM tied to functional consequence (mineralization); single lab\",\n      \"pmids\": [\"36361965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"miR-24-3p directly binds the 3'UTR of ANXA6 mRNA and suppresses ANXA6 expression, thereby promoting proliferation and inhibiting adipogenic differentiation of chicken intramuscular preadipocytes. Overexpression of ANXA6 inhibited proliferation and promoted differentiation, while ANXA6 knockdown had opposite effects. Rescue with an ANXA6 overexpression vector eliminated the impact of miR-24-3p mimics, confirming ANXA6 as the functional mediator.\",\n      \"method\": \"Dual-luciferase reporter assay for miR-24-3p binding to ANXA6 3'UTR, ANXA6 overexpression and knockdown, miR-24-3p mimics/inhibitors, cell proliferation and adipogenic differentiation assays\",\n      \"journal\": \"Genes\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — dual-luciferase validates direct miRNA-target interaction; rescue experiment confirms functional axis; single lab\",\n      \"pmids\": [\"35456441\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"AnxA6 is SUMOylated by SUMO1, with K299 identified as a key SUMOylation site. SUMOylated AnxA6 preferentially binds EGFR upon EGF stimulation and facilitates EGFR-PKCα complex formation, which decreases EGF-induced phosphorylation of EGFR and ERK1/2 and reduces cyclin D1 expression. The SUMO-site mutant AnxA6K299R showed reduced suppression of EGFR-ERK1/2 phosphorylation, reduced tumor suppression, and conferred gefitinib resistance. This mechanism also operates on mutant EGFR (T790M/L858R).\",\n      \"method\": \"Co-transfection with SUMO1/UBC9 plasmids, immunoprecipitation for SUMOylated AnxA6, LC-MS/MS for SUMOylation site mapping, site-directed mutagenesis (K299R), shRNA knockdown, lentiviral overexpression, Western blot, CCK8/colony/wound healing assays, xenograft nude mouse model\",\n      \"journal\": \"Cell communication and signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — SUMOylation site identified by MS and validated by mutagenesis, functional consequences confirmed in vitro and in vivo; multiple orthogonal methods\",\n      \"pmids\": [\"37528485\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The ANXA6/TRPV2 axis promotes lymphatic metastasis in head and neck squamous cell carcinoma by inducing autophagy through inhibition of the AKT/mTOR signaling pathway. ANXA6 expression positively correlated with TRPV2 expression, and TRPV2 inhibition reversed ANXA6-induced autophagy and lymphatic metastasis, placing TRPV2 downstream of ANXA6 in this pathway.\",\n      \"method\": \"RNA sequencing of clinical specimens, ANXA6 overexpression/knockdown, in vitro proliferation and migration assays, in vivo lymphatic metastasis assays, TRPV2 inhibitor treatment, AKT/mTOR pathway Western blot\",\n      \"journal\": \"Experimental hematology & oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic and pharmacological epistasis with defined pathway readouts in vitro and in vivo; single lab\",\n      \"pmids\": [\"37138336\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PRRX1 was identified as a transcription factor that directly promotes ANXA6 expression in bladder cancer via ChIP-PCR and dual-luciferase assays. ANXA6 in turn promotes PKCα/EGFR complex formation (confirmed by mass spectrometry, immunofluorescence, and co-immunoprecipitation), inhibiting EGFR phosphorylation and downstream AKT and ERK1/2 signaling, thereby enhancing cisplatin sensitivity and re-sensitizing resistant cells.\",\n      \"method\": \"ChIP-PCR, dual-luciferase reporter assay for PRRX1 binding to ANXA6 promoter, mass spectrometry, immunofluorescence, co-immunoprecipitation, CCK-8, colony formation, flow cytometry (ROS, apoptosis), comet assay, xenograft model\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (ChIP, Co-IP, MS) confirming transcriptional regulation and downstream complex; single lab\",\n      \"pmids\": [\"39528080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Anxa6-/- mice display rapid fasting-induced hypoglycemia due to a specific impairment in the glucose-alanine cycle: these mice cannot utilize alanine for hepatic gluconeogenesis (GNG). Fasted Anxa6-/- mice showed elevated glucagon, increased lipid oxidation, and slightly reduced expression of alanine aminotransferase 2 (Gpt2), lactate dehydrogenase (Ldha2), and the hepatic alanine transporter SNAT4, demonstrating a critical role for ANXA6 in alanine-dependent hepatic GNG.\",\n      \"method\": \"Anxa6-/- knockout mouse model, indirect calorimetry, glucose/insulin/glucagon tolerance tests, alanine challenge tests, respiratory exchange ratio measurement, biochemical enzyme expression analysis\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO mouse with specific metabolic challenge revealing alanine-GNG defect; single lab\",\n      \"pmids\": [\"40802799\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ANXA6 overexpression in decidual macrophages promotes macrophage M1 polarization and pyroptosis through the NF-κB signaling pathway, leading to mitochondrial ROS accumulation and decreased mitochondrial membrane potential. Inhibition of NLRP3 rescued ANXA6-overexpression-induced M1 polarization and pyroptosis. ANXA6-overexpressing macrophages inhibited trophoblast function in vitro via TNF-α-mediated inhibition of the PI3K/AKT axis.\",\n      \"method\": \"ANXA6 overexpression in macrophages, ROS/mitochondrial membrane potential assays, NLRP3 inhibitor treatment, ROS inhibitor treatment, NF-κB pathway analysis, M1/M2 polarization markers, trophoblast co-culture, PI3K/AKT pathway Western blot\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological and genetic epistasis across multiple pathways; single lab\",\n      \"pmids\": [\"40959268\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MG53 binds AnxA6 at membrane injury sites, and AnxA6 overexpression abolishes MG53's cytoprotective effects in iPSCs under hypoxia/reoxygenation stress, placing AnxA6 as a negative regulator of MG53-mediated membrane repair. MG53 treatment enhanced iPSC survival and functional integration in myocardial tissue post-ischemia-reperfusion injury in vivo.\",\n      \"method\": \"Co-binding assay/interaction studies (MG53-AnxA6), AnxA6 overexpression rescue experiment, H/R injury model in iPSCs, in vivo MIRI model with MG53 treatment and iPSC transplantation\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — interaction inferred from functional rescue; single method for binding, single lab\",\n      \"pmids\": [\"41106752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ANXA6 co-localizes with cGAS and acts as an inhibitor of the cGAS-STING signaling pathway by promoting K63-linked ubiquitination of cGAS, thereby reducing cGAS protein levels. ANXA6 knockdown impeded cGAS K63-ubiquitination, increased cGAS expression, activated downstream TBK1, IRF3, and type I IFN-stimulated genes (MX1, ISG15, ISG56), elevated ROS and lipid peroxides, and induced ferroptosis in cervical cancer cells.\",\n      \"method\": \"Co-localization by immunofluorescence, co-immunoprecipitation for ANXA6-cGAS interaction, ubiquitination assays (K63-linkage specificity), ANXA6 siRNA knockdown, Western blot for cGAS-STING pathway components, ROS/lipid peroxide assays, ferroptosis detection\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP plus ubiquitination assay with downstream pathway readouts; single lab\",\n      \"pmids\": [\"41219880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In DMD skeletal muscle cells, ANXA5 and ANXA6 fail to upregulate in response to mechanical membrane stress, while control cells upregulate both proteins as part of an adaptive membrane repair mechanism. This was established by live-cell imaging of GFP-tagged annexins, Western blotting, and a shear stress-based injury assay, placing ANXA6 as part of the adaptive membrane repair response in skeletal muscle.\",\n      \"method\": \"Live-cell imaging of GFP-tagged ANXA6, shear stress-based membrane injury assay, Western blot for annexin expression, immunohistochemistry of DMD patient biopsies\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — direct localization by live imaging tied to membrane repair function; preprint, single lab\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"AnxA6 interacts with SNAP23 (a SNARE protein) and this interaction is required for the translocation of AnxA6 to cellular membranes and subsequent secretion of pro-inflammatory cytokines (MCP-1/CCL2, IL-8, DKK1, TSP-1, osteopontin) and small extracellular vesicles (EVs) in triple-negative breast cancer cells. AnxA6 downregulation strongly reduced EV secretion and cytokine levels; upregulation promoted EV-associated Rab7, cholesterol, and MCP-1. AnxA6 neutralizing antibodies reduced survival of AnxA6-low TNBC cells.\",\n      \"method\": \"Co-immunoprecipitation (AnxA6-SNAP23), cytokine arrays on conditioned media, EV isolation and characterization, AnxA6 siRNA knockdown and overexpression, cholesterol measurement in EVs, membrane translocation assays\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP for SNAP23 interaction, combined with functional EV and cytokine secretion assays; preprint, single lab\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"ANXA6 is a Ca2+-dependent, phospholipid-binding scaffold protein that: (1) inhibits membrane fusion in a Ca2+-dependent manner and partitions into cholesterol/sphingomyelin-enriched microdomains; (2) suppresses EGFR-ERK1/2 signaling by facilitating EGFR-PKCα complex formation, an activity enhanced by SUMO1 modification at K299; (3) promotes autophagy by inhibiting the PI3K/AKT/mTOR pathway and enabling ATG9A vesicle trafficking via RAB GTPases/F-actin; (4) functions as a scaffold for the ANXA6/LRP1/TSP1 complex in cancer-associated fibroblast-derived EVs that promote pancreatic cancer aggressiveness; (5) acts as a host restriction factor for influenza A virus; (6) is required for hepatic alanine-dependent gluconeogenesis; (7) inhibits the cGAS-STING pathway by promoting K63-ubiquitination of cGAS; and (8) facilitates secretion of pro-inflammatory cytokines and EVs through interaction with SNAP23.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"ANXA6 is a Ca²⁺-dependent phospholipid-binding scaffold that negatively regulates growth factor signaling and modulates membrane-associated trafficking events. ANXA6 suppresses EGFR-ERK1/2 signaling by facilitating PKCα–EGFR complex formation in a manner dependent on SUMO1 modification at K299, and independently inhibits the PI3K–AKT–mTOR axis to promote autophagy and ATG9A-positive vesicle trafficking from recycling endosomes to autophagosomes [PMID:37528485, PMID:33135350, PMID:32373608]. ANXA6 interacts with cGAS to promote its ubiquitination and suppress cGAS-STING innate immune signaling [PMID:41219880], and functions as a host restriction factor for influenza A virus replication [PMID:34673207]. In hepatic metabolism, Anxa6 knockout mice display impaired alanine-dependent gluconeogenesis and fasting hypoglycemia, establishing ANXA6 as a regulator of hepatic glucose production [PMID:40802799, PMID:30110341].\",\n  \"teleology\": [\n    {\n      \"year\": 2013,\n      \"claim\": \"Establishing that ANXA6 membrane association is primarily Ca²⁺-driven answered how ANXA6 is recruited to specific lipid microdomains, showing Ca²⁺-dependent translocation to cholesterol/sphingomyelin-enriched detergent-resistant membranes.\",\n      \"evidence\": \"Langmuir monolayer reconstitution with DPPC/sphingomyelin and sucrose gradient DRM fractionation from Niemann-Pick C cells\",\n      \"pmids\": [\"23360953\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural basis for lipid selectivity\", \"Role of cholesterol accumulation (NPC context) versus normal physiology unclear\", \"Ca²⁺ threshold for translocation not quantified in vivo\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Demonstrating that ANXA6 is required for cancer cell invasion via antibody blocking and siRNA knockdown established ANXA6 as a functionally relevant cell-surface factor in tumor invasiveness.\",\n      \"evidence\": \"Anti-AnxA6 MAb 9E1 blocking and siRNA silencing with Matrigel invasion assays in pancreatic, lung, and breast cancer cells; target confirmed by IP-LC-MS/MS\",\n      \"pmids\": [\"28881357\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism linking cell-surface ANXA6 to invasion not identified\", \"Downstream effectors of pro-invasive activity unknown\", \"In vivo metastasis validation absent\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Anxa6 knockout mice revealed a paradoxical metabolic phenotype — enhanced hepatic insulin signaling (increased Akt/S6K phosphorylation) but failure to suppress gluconeogenesis — establishing ANXA6 as a regulator of hepatic glucose metabolism beyond canonical insulin-Akt output.\",\n      \"evidence\": \"Anxa6-KO mice on HFD with OGTT/ITT, hepatocyte glucose production assays, Western blot for Akt/S6K\",\n      \"pmids\": [\"30110341\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct substrate or binding partner mediating gluconeogenic regulation unknown\", \"Mechanism uncoupling Akt activation from gluconeogenesis suppression not resolved\", \"Contribution of extrahepatic ANXA6 not excluded in whole-body KO\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Two studies converged on ANXA6 as a negative regulator of PI3K-AKT-mTOR signaling that promotes autophagy and ATG9A-positive vesicle trafficking, resolving how ANXA6 links membrane scaffolding to autophagosome biogenesis.\",\n      \"evidence\": \"siRNA knockdown and restoration in NPC and other cells; CAL101 epistasis; ATG9A trafficking from recycling endosomes; RAB GTPase interactome analysis; LC3 readouts\",\n      \"pmids\": [\"32373608\", \"33135350\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct physical interaction with PI3K or mTOR complexes not demonstrated\", \"Which RAB GTPases are direct versus indirect ANXA6 partners unclear\", \"Whether autophagy role is Ca²⁺-dependent not tested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identification of ANXA6 as a direct binding target of bacterial effector EspF linked ANXA6 to tight junction regulation through MLCK/MLC/PKCα activation, revealing a pathogen exploitation mechanism.\",\n      \"evidence\": \"FRET and co-IP for EspF–ANXA6 interaction; Western blot for ZO-1, occludin, phospho-MLC, PKCα\",\n      \"pmids\": [\"33425920\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ANXA6 has an endogenous role in tight junction maintenance independent of EspF unknown\", \"Structural basis of EspF–ANXA6 interaction not determined\", \"In vivo intestinal barrier phenotype not tested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"CRISPR knockout of ANXA6 increased influenza A virus replication, establishing ANXA6 as a host restriction factor and raising the question of which step of the viral life cycle is inhibited.\",\n      \"evidence\": \"CRISPR-Cas9 KO in HEK293FT cells with TCID assay and viral growth curves\",\n      \"pmids\": [\"34673207\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of antiviral restriction (entry, replication, assembly, or budding) not defined\", \"Whether other annexins compensate in vivo not addressed\", \"Rescue with ANXA6 re-expression not shown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identification of K299 SUMOylation by SUMO1 as the modification that enables ANXA6 to bind EGFR and scaffold the PKCα–EGFR inhibitory complex resolved how ANXA6 suppresses EGFR-ERK1/2 signaling at a molecular level.\",\n      \"evidence\": \"LC-MS/MS SUMOylation site mapping, K299R mutagenesis abolishing EGFR-ERK suppression, Co-IP for EGFR-PKCα complex, xenograft tumor suppression\",\n      \"pmids\": [\"37528485\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of SUMO E3 ligase for ANXA6 unknown\", \"Whether SUMOylation also regulates autophagy or other ANXA6 functions not tested\", \"Structural basis of SUMO-dependent EGFR binding not resolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Discovery that ANXA6 binds cGAS and promotes its ubiquitination to suppress cGAS-STING signaling expanded ANXA6's role to innate immune regulation, with downstream effects on TBK1-IRF3 activation and ferroptosis.\",\n      \"evidence\": \"Co-localization and co-IP for ANXA6–cGAS interaction; ubiquitination assays showing K63-linked ubiquitin modulation; KD with phospho-TBK1/IRF3/ISG readouts\",\n      \"pmids\": [\"41219880\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ANXA6 is the E3 ligase or recruits one is unknown\", \"Specificity for K63-linked versus other ubiquitin chain types needs independent confirmation\", \"Whether Ca²⁺ binding is required for cGAS interaction not tested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identification of PRRX1 as a transcriptional activator of ANXA6 and confirmation of PKCα/EGFR complex-mediated cisplatin sensitization in bladder cancer independently validated the EGFR-suppressive mechanism and connected ANXA6 to chemoresistance.\",\n      \"evidence\": \"ChIP-PCR and dual-luciferase for PRRX1-ANXA6 promoter binding; Co-IP for PKCα-EGFR; xenograft cisplatin response\",\n      \"pmids\": [\"39528080\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether PRRX1 regulation is tissue-specific unknown\", \"Relationship between PRRX1-driven ANXA6 expression and SUMOylation status not explored\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Refined metabolic phenotyping of Anxa6-KO mice pinpointed alanine-dependent gluconeogenesis as the specific pathway disrupted, explaining fasting hypoglycemia through reduced Gpt2, Ldha2, and SNAT4 expression.\",\n      \"evidence\": \"Anxa6-KO mice with indirect calorimetry, alanine tolerance tests, enzyme/transporter expression analysis\",\n      \"pmids\": [\"40802799\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ANXA6 directly regulates Gpt2/SNAT4 transcription or acts indirectly unknown\", \"Liver-specific versus systemic KO phenotype not distinguished\", \"Human relevance of alanine gluconeogenesis defect not established\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include: the structural basis for how SUMOylation at K299 alters ANXA6 protein–protein interactions; whether Ca²⁺-dependent membrane recruitment is required for each of ANXA6's distinct signaling functions; and the identity of E3 ligases mediating ANXA6-promoted cGAS ubiquitination.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No crystal structure of SUMOylated ANXA6 or ANXA6-EGFR complex\", \"Unified model linking Ca²⁺ binding, membrane translocation, and signaling suppression across different pathways absent\", \"In vivo immune phenotype of ANXA6 loss not characterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [8, 12]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [2, 3, 10]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 2, 3, 11]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [8, 12, 16]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1, 2]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 2, 3, 11]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [0, 1, 11]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [5, 10]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [6, 7]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"EGFR\",\n      \"PRKCA\",\n      \"CGAS\",\n      \"ATG9A\",\n      \"SNAP23\",\n      \"MG53\",\n      \"PRRX1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"ANXA6 is a Ca²⁺-dependent phospholipid-binding scaffold protein that organizes membrane-associated signaling platforms, regulates vesicular trafficking and autophagy, and modulates metabolic and innate immune pathways. Its eight-repeat annexin fold binds phosphatidylserine, phosphatidic acid, phosphatidylinositol, and phosphatidylethanolamine in a Ca²⁺-dependent manner and inhibits phospholipid vesicle fusion [PMID:2138016]; Ca²⁺ also drives its partitioning into cholesterol/sphingomyelin-enriched membrane microdomains [PMID:23360953]. ANXA6 suppresses EGFR–ERK1/2 signaling by facilitating PKCα–EGFR complex formation, an activity enhanced by SUMO1 modification at K299, and loss of this SUMOylation confers gefitinib resistance [PMID:37528485, PMID:39528080]. ANXA6 additionally promotes autophagy through dual inhibition of PI3K/AKT/mTOR signaling and facilitation of ATG9A vesicle trafficking via RAB GTPases/F-actin [PMID:32373608, PMID:33135350], functions as a host restriction factor for influenza A virus [PMID:34673207], is required for hepatic alanine-dependent gluconeogenesis [PMID:40802799], and inhibits the cGAS–STING pathway by promoting K63-linked ubiquitination of cGAS [PMID:41219880].\",\n  \"teleology\": [\n    {\n      \"year\": 1988,\n      \"claim\": \"Determining the primary structure of ANXA6 established it as a Ca²⁺-binding annexin family member with an unusual eight-repeat architecture, providing the molecular framework for all subsequent functional studies.\",\n      \"evidence\": \"cDNA cloning and sequence analysis of human p68/ANXA6\",\n      \"pmids\": [\"3258820\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No three-dimensional structure resolved at this stage\",\n        \"Biological function and lipid-binding specificity not yet characterized\"\n      ]\n    },\n    {\n      \"year\": 1990,\n      \"claim\": \"Reconstitution with defined lipid vesicles revealed that ANXA6 uniquely inhibits membrane fusion at threshold Ca²⁺ and binds selectively to anionic phospholipids, distinguishing its biophysical activity from other annexins.\",\n      \"evidence\": \"In vitro phospholipid vesicle aggregation and fusion assays with purified ANXA6\",\n      \"pmids\": [\"2138016\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Cellular consequences of fusion inhibition not demonstrated\",\n        \"Whether ANXA6 inhibits fusion in vivo remains untested\"\n      ]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Showing that Ca²⁺ drives ANXA6 into cholesterol/sphingomyelin-enriched microdomains connected its lipid-binding activity to raft-dependent signaling and linked it mechanistically to Niemann-Pick type C pathology.\",\n      \"evidence\": \"Detergent-resistant membrane fractionation and Langmuir monolayer assays in NPC cells\",\n      \"pmids\": [\"23360953\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether ANXA6 raft partitioning is causative versus correlative in NPC disease\",\n        \"Structural basis for cholesterol/sphingomyelin selectivity not resolved\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identification of a stable ANXA6/LRP1/TSP1 complex in cancer-associated fibroblast-derived extracellular vesicles established ANXA6 as a scaffolding component of pro-tumorigenic paracrine signaling in pancreatic cancer.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation, EV uptake, ANXA6 depletion in CAFs, in vivo xenograft and metastasis assays\",\n      \"pmids\": [\"27701147\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Mechanism by which ANXA6 recruits LRP1 and TSP1 into EVs is unknown\",\n        \"Direct stoichiometry and structure of the ternary complex not determined\"\n      ]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Anxa6-knockout mice revealed an unexpected metabolic role: despite intact insulin–Akt signaling, ANXA6 loss uncouples insulin action from suppression of hepatic gluconeogenesis, positioning ANXA6 as a fine-tuning scaffold for hepatic glucose metabolism.\",\n      \"evidence\": \"Anxa6-KO mouse model with high-fat diet, glucose/insulin tolerance tests, Akt/S6K phosphorylation, glucose production assay in AnxA6-depleted hepatocytes\",\n      \"pmids\": [\"30110341\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Direct protein target through which ANXA6 controls gluconeogenesis downstream of Akt not identified\",\n        \"Whether phenotype is liver-autonomous versus systemic not fully resolved\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Two independent studies converged to show ANXA6 promotes autophagy through dual mechanisms—inhibition of PI3K/AKT/mTOR signaling and facilitation of ATG9A vesicle trafficking via RAB GTPases and F-actin—resolving how ANXA6 connects membrane dynamics to autophagosome biogenesis.\",\n      \"evidence\": \"ANXA6 knockdown/rescue with LC3 and ATG9A trafficking readouts, epistasis with PI3K inhibitor CAL101, mTOR/ERK pathway analysis\",\n      \"pmids\": [\"32373608\", \"33135350\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Which RAB GTPases directly interact with ANXA6 for ATG9A sorting not defined\",\n        \"Structural basis for ANXA6 engagement with autophagic machinery unknown\",\n        \"Whether the PI3K/AKT arm and the ATG9A trafficking arm are independent or coupled is unclear\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"CRISPR knockout demonstrated that endogenous ANXA6 restricts influenza A virus replication, expanding its functional repertoire to innate antiviral defense.\",\n      \"evidence\": \"CRISPR-Cas9 ANXA6 KO in HEK293FT cells, TCID50 viral titer and growth curve assays\",\n      \"pmids\": [\"34673207\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism of viral restriction (virion packaging, entry, or egress) not delineated\",\n        \"Relevance in primary airway epithelial cells not tested\",\n        \"Whether ANXA6 restricts other viruses is unknown\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identification of K299 as a SUMO1 modification site explained how ANXA6 suppresses EGFR–ERK1/2 signaling: SUMOylated ANXA6 bridges EGFR and PKCα, and loss of this modification confers resistance to EGFR-targeted therapy, unifying the tumor-suppressive and drug-sensitizing activities of ANXA6.\",\n      \"evidence\": \"LC-MS/MS SUMOylation site mapping, K299R mutagenesis, co-IP for EGFR–PKCα complex, in vitro and xenograft assays\",\n      \"pmids\": [\"37528485\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"E3 SUMO ligase responsible for K299 modification not identified\",\n        \"Whether SUMOylation modulates ANXA6 lipid-binding or raft partitioning is untested\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Anxa6-KO mice challenged with alanine revealed a specific requirement for ANXA6 in the glucose–alanine cycle, pinpointing the metabolic defect to alanine-dependent hepatic gluconeogenesis and linking ANXA6 to SNAT4/Gpt2 expression.\",\n      \"evidence\": \"Anxa6-KO mouse model, alanine challenge, indirect calorimetry, transporter and enzyme expression analysis\",\n      \"pmids\": [\"40802799\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether ANXA6 directly regulates SNAT4 trafficking or Gpt2 transcription is unknown\",\n        \"Tissue-specific rescue experiments not reported\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstration that ANXA6 promotes K63-linked ubiquitination of cGAS to suppress the cGAS–STING–IFN axis established a new role for ANXA6 in tuning innate immune DNA sensing, with consequences for ferroptosis induction when ANXA6 is lost.\",\n      \"evidence\": \"Co-IP for ANXA6–cGAS, K63-ubiquitination assay, ANXA6 knockdown with STING pathway and ferroptosis readouts in cervical cancer cells\",\n      \"pmids\": [\"41219880\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"E3 ubiquitin ligase recruited by ANXA6 to ubiquitinate cGAS not identified\",\n        \"Whether ANXA6 directly binds cGAS or acts via an intermediate is unresolved\",\n        \"In vivo validation of the cGAS–STING suppressive role is lacking\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The upstream signals that coordinate ANXA6's scaffolding functions across its diverse pathways—EGFR suppression, autophagy, gluconeogenesis, cGAS regulation, and membrane repair—remain mechanistically unintegrated; a unifying structural or regulatory model is absent.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No full-length ANXA6 structure in complex with any signaling partner exists\",\n        \"How SUMOylation, Ca²⁺ binding, and lipid raft partitioning are coordinately regulated is unknown\",\n        \"Tissue-specific isoform functions arising from alternative splicing remain largely unexplored\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [1, 2]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 13, 15]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [6, 8, 13, 19]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 2, 11, 21]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [3, 8, 9]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [13, 19]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [3, 9, 21]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [6, 8, 13, 14, 15]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [6, 8, 9, 14]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [3, 9, 21]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [5, 16]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [10, 19]}\n    ],\n    \"complexes\": [\n      \"ANXA6/LRP1/TSP1 (CAF-derived EV complex)\",\n      \"EGFR/PKCα/ANXA6 complex\"\n    ],\n    \"partners\": [\n      \"EGFR\",\n      \"PRKCA\",\n      \"LRP1\",\n      \"THBS1\",\n      \"SNAP23\",\n      \"CGAS\",\n      \"ATG9A\",\n      \"TRIM72\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}