{"gene":"ADCY9","run_date":"2026-06-09T22:02:41","timeline":{"discoveries":[{"year":1998,"finding":"Human ADCY9 (AC9) is stimulated by beta-adrenergic receptor activation but is insensitive to forskolin, Ca2+, and somatostatin when expressed in HEK-293 cells; unlike mouse AC9, human AC9 activity is unaffected by inhibitors of calcineurin. Divergence at the C2a/C2b junction results in an alternative C2b amino acid sequence compared to mouse AC9.","method":"Heterologous expression in HEK-293 cells, pharmacological assays, cAMP measurement","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct functional assay in mammalian cells with multiple pharmacological probes, single lab","pmids":["9628827"],"is_preprint":false},{"year":2008,"finding":"miR-142-3p targets AC9 mRNA to suppress cAMP production in CD4+CD25- T cells; in CD4+CD25+ T regulatory cells, FOXP3 downregulates miR-142-3p, thereby keeping the AC9/cAMP pathway active and enabling high cAMP levels required for suppressor function.","method":"miRNA target validation (luciferase reporter, mRNA/protein quantification), T cell functional assays, cAMP measurement, FOXP3 overexpression/knockdown","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal regulation validated with multiple methods (reporter assay, gain/loss of function, functional T cell assays), replicated across two cell types","pmids":["19098714"],"is_preprint":false},{"year":2013,"finding":"miR-181b targets AC9 mRNA to restrict cAMP production post-transcriptionally in cervical cancer cells; knockdown of AC9 phenocopies miR-181b overexpression (increased proliferation, reduced apoptosis), while AC9 overexpression has opposite effects.","method":"miRNA target validation (luciferase reporter assay), siRNA knockdown and overexpression, cAMP measurement, proliferation and apoptosis assays","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reporter assay plus functional rescue experiments, single lab","pmids":["24269684"],"is_preprint":false},{"year":2014,"finding":"AC9 catalyzes cAMP production in APL cells and enhances retinoic acid receptor trans-activity; AC9 knockdown inhibits ATRA-induced differentiation of NB4 cells. miR-181a targets the 3'UTR of AC9 mRNA to decrease AC9 expression and intracellular cAMP, and CEBPα inhibits miR-181a expression, accounting for differential AC9 levels between ATRA-sensitive and resistant cells.","method":"Luciferase reporter assay (3'UTR targeting), siRNA knockdown, overexpression, cAMP measurement, differentiation assays, CEBPα gain/loss of function","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (reporter, KD, OE, functional readout), single lab","pmids":["24722286"],"is_preprint":false},{"year":2018,"finding":"The isoform-specific carboxyl-terminal C2b domain of AC9 auto-inhibits enzyme activation by Gs-coupled receptors; deletion of the C2b domain markedly enhances cAMP response to beta-adrenergic and prostanoid receptor activation. Residues 1268-1276 within C2b are critical for auto-inhibition. Two molecular weight species of AC9 (~130K and ≥170K) are detected in myocardial membranes, with the lower species lacking the C2b domain, suggesting proteolytic cleavage may regulate AC9 activity.","method":"Deletion mutagenesis, stable overexpression in HEK-293 cells, cAMP assay, immunoblotting with domain-specific antibodies, rodent and human myocardial membrane fractionation","journal":"Cellular signalling","confidence":"High","confidence_rationale":"Tier 1 / Moderate — mutagenesis (deletion and point mutants) with direct enzymatic readout plus tissue biochemistry, single lab with multiple orthogonal approaches","pmids":["30121334"],"is_preprint":false},{"year":2018,"finding":"Adcy9 inactivation in mice reduces aortic atherosclerosis by 65%, decreases macrophage accumulation and proliferation in plaques, and improves endothelial-dependent vasorelaxation via nitric oxide, cyclooxygenase, and endothelial-dependent hyperpolarization pathways. These protective effects are entirely lost when CETP is present (CETPtg Adcy9Gt/Gt mice), demonstrating that Adcy9 inactivation protects from atherosclerosis only in the absence of CETP activity.","method":"Genetic inactivation (Adcy9Gt/Gt mice), atherogenic diet protocol, histology (CD68 macrophage staining), vasorelaxation pharmacology, telemetry, adipose tissue MRI, splenocyte adhesion assay","journal":"Circulation","confidence":"High","confidence_rationale":"Tier 2 / Strong — comprehensive in vivo loss-of-function with multiple orthogonal readouts (atherosclerosis, endothelial function, macrophage biology, autonomic tone), epistasis with CETP transgene","pmids":["29674325"],"is_preprint":false},{"year":2019,"finding":"miR-142-3p targets AC9 in dorsal root ganglion/spinal cord neurons; miR-142-3p inhibition upregulates AC9, elevates cAMP and downstream p-CREB, IL-6, and GAP43, and reduces GTP-RhoA, promoting axon growth in vitro and sensory conduction recovery in vivo after dorsal column injury. Co-knockdown of AC9 reverses these effects. Sorafenib mimics SNCI by downregulating miR-142-3p.","method":"miR-142-3p inhibitor/mimic delivery in rats, AC9 siRNA, axon length measurement, cAMP/protein quantification, in vivo sensory conduction electrophysiology","journal":"Neuropharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis (miR inhibitor + AC9 siRNA rescue), in vitro and in vivo functional readouts, single lab","pmids":["30710569"],"is_preprint":false},{"year":2020,"finding":"miR-142-3p directly targets AC9 (validated by luciferase reporter assay) in sciatic nerve injury (CCI) rats; miR-142-3p expression increases while AC9 and cAMP decrease in CCI rats. miR-142-3p silencing reduces neuropathic pain and inflammatory cytokines via upregulation of AC9 and downstream cAMP/AMPK pathway proteins.","method":"Dual luciferase reporter assay, miR-142-3p mimic/siRNA delivery in CCI rats, AC9 overexpression/siRNA, cAMP measurement, AMPK pathway protein quantification, pain behavioral testing","journal":"International journal of molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — luciferase validation of targeting plus in vivo epistasis experiments, single lab","pmids":["33416140"],"is_preprint":false},{"year":2020,"finding":"Deletion of adcy9 in zebrafish (morphant model) causes cardiac malformation with increased macrophage migration, cardiac apoptosis, and upregulation of mmp9 (matrix metalloproteinase involved in extracellular matrix remodeling), supporting ADCY9 as a candidate gene for congenital heart defects.","method":"Zebrafish adcy9 morpholino knockdown, immunofluorescence (macrophage markers), RNA sequencing, cardiac morphology assessment","journal":"Orphanet journal of rare diseases","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo loss-of-function with immunofluorescence and transcriptomics, single lab, zebrafish model","pmids":["32321550"],"is_preprint":false},{"year":2022,"finding":"miR-210-3p targets ADCY9 mRNA (validated by target relation detection); miR-210-3p is downregulated while ADCY9 is upregulated in myocardial infarction rats. Sevoflurane-mediated cardioprotection (reduced injury, fibrosis, and apoptosis) is potentiated by miR-210-3p elevation and abolished by miR-210-3p inhibition or ADCY9 overexpression, placing ADCY9 downstream of miR-210-3p in the sevoflurane cardioprotective pathway.","method":"miR-210-3p agomir/antagomir delivery, ADCY9 overexpression in MI rat model, echocardiography, histology, miRNA-target validation assay","journal":"Functional & integrative genomics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — functional rescue experiments in vivo but target validation method not fully described in abstract, single lab","pmids":["34988676"],"is_preprint":false},{"year":2023,"finding":"Adcy9 inactivation in mice after myocardial infarction reduces infarct size, pathological left ventricular remodeling, and cardiac dysfunction, and preserves myocardial capillary density in the infarct border zone; increased bone marrow T cells and B cells were also observed. These benefits were only seen in the absence of CETP activity.","method":"Adcy9Gt/Gt mice, permanent LAD ligation MI model, echocardiography, histology (cardiomyocyte size, infarct size, capillary density), flow cytometry (immune cells)","journal":"The Canadian journal of cardiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo genetic loss-of-function with multiple histological and functional readouts, CETP epistasis, single lab","pmids":["37054880"],"is_preprint":false},{"year":2023,"finding":"ADCY9 overexpression in lung adenocarcinoma cell lines (SPCA1, A549) restrains cell proliferation, invasion, and migration, consistent with a tumor-suppressive function mediated through the cAMP pathway.","method":"ADCY9 overexpression in cancer cell lines, proliferation assay, invasion/migration assays","journal":"Journal of thoracic disease","confidence":"Low","confidence_rationale":"Tier 3 / Weak — overexpression phenotype assays only, no direct mechanistic pathway validation, single lab","pmids":["37065546"],"is_preprint":false},{"year":2025,"finding":"Asthma-related cytokines decrease ADCY9 expression and cellular cAMP levels in airway smooth muscle cells (ASMCs), reducing airway relaxation and promoting remodeling. ADCY9 overexpression attenuates airway smooth muscle remodeling, but the missense variant ADCY9-Ile772Met (rs2230739) fails to prevent remodeling, demonstrating that the Ile772Met substitution impairs ADCY9's protective function in the airway.","method":"ADCY9 overexpression and mutant (Ile772Met) expression in ASMCs, cAMP measurement, airway remodeling assays, FEV1/FVC% clinical correlation","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional comparison of wild-type vs missense mutant with enzymatic (cAMP) and cellular readouts, single lab","pmids":["40816614"],"is_preprint":false},{"year":2027,"finding":"In planarian (Dugesia japonica), ADCY9 knockdown causes abnormal brain regeneration (loss of nerve cord, reduced collateral branches, inhibited neuron regeneration/differentiation). Double RNAi of ADCY9 and Mitofusin-1 restores neural regeneration, indicating ADCY9 promotes neural regeneration by negatively regulating the downstream inhibitory factor Mitofusin-1 in the cAMP/AMPK signaling pathway.","method":"RNAi knockdown (single and double) in planarians, RNA sequencing, KEGG pathway analysis, morphological assessment of brain regeneration","journal":"Cells","confidence":"Low","confidence_rationale":"Tier 3 / Weak — genetic epistasis by double RNAi in planarian model, single lab, no direct biochemical validation of ADCY9-Mitofusin-1 interaction","pmids":["41827823"],"is_preprint":false}],"current_model":"ADCY9 (AC9) is a transmembrane adenylyl cyclase that catalyzes ATP-to-cAMP conversion and is activated by Gs-coupled receptors (including beta-adrenergic receptors) but is insensitive to forskolin and Ca2+; its activity is auto-inhibited by an isoform-specific C2b carboxyl-terminal domain (residues 1268-1276), potentially relieved by proteolytic cleavage; ADCY9 is post-transcriptionally suppressed by multiple miRNAs (miR-142-3p, miR-181a, miR-181b, miR-210-3p) that target its 3'UTR to reduce intracellular cAMP, with FOXP3 counter-regulating miR-142-3p in T regulatory cells to maintain high cAMP; in vivo, ADCY9 inactivation protects against atherosclerosis, reduces myocardial infarction injury, and improves endothelial function exclusively in the absence of CETP activity, revealing a genetic epistatic interaction between ADCY9 and CETP."},"narrative":{"mechanistic_narrative":"ADCY9 (AC9) is a transmembrane adenylyl cyclase that synthesizes cAMP downstream of Gs-coupled receptors, including beta-adrenergic and prostanoid receptors, and is distinguished from other adenylyl cyclases by its insensitivity to forskolin and Ca2+ [PMID:9628827]. Its activation is restrained by an isoform-specific carboxyl-terminal C2b domain (residues 1268-1276), whose deletion markedly amplifies the cAMP response to receptor stimulation; two molecular-weight AC9 species in myocardial membranes indicate that proteolytic removal of C2b can relieve this auto-inhibition [PMID:30121334]. ADCY9 output is set post-transcriptionally by multiple miRNAs that target its 3'UTR—miR-142-3p, miR-181a/b, and miR-210-3p—each lowering AC9 protein and intracellular cAMP, with upstream regulators such as FOXP3 and CEBPalpha tuning these miRNAs to control cAMP in T regulatory cells and during myeloid differentiation [PMID:19098714, PMID:24722286]. Through this cAMP-generating activity, ADCY9 governs diverse physiological outputs: it sustains regulatory T cell suppressor function [PMID:19098714], supports retinoic-acid-induced differentiation [PMID:24722286], and shapes cardiovascular biology, where genetic Adcy9 inactivation reduces atherosclerosis, limits myocardial infarction injury, and improves endothelial function—effects observed exclusively in the absence of CETP activity, defining a genetic epistasis between ADCY9 and CETP [PMID:29674325, PMID:37054880]. In airway smooth muscle, ADCY9-driven cAMP promotes relaxation and opposes remodeling, a protective function abolished by the missense variant Ile772Met [PMID:40816614].","teleology":[{"year":1998,"claim":"Established that human AC9 is a functional adenylyl cyclase coupled to Gs receptors yet pharmacologically atypical, defining its identity among the cyclase family.","evidence":"Heterologous expression in HEK-293 cells with beta-adrenergic, forskolin, Ca2+ and calcineurin probes plus cAMP measurement","pmids":["9628827"],"confidence":"Medium","gaps":["No structural basis for forskolin insensitivity defined","Endogenous receptor partners in native tissue not mapped"]},{"year":2008,"claim":"Showed that AC9 is post-transcriptionally controlled by miR-142-3p, and that FOXP3 relieves this repression to maintain the high cAMP required for Treg suppressor function, linking AC9 to immune regulation.","evidence":"Luciferase reporter, mRNA/protein quantification, FOXP3 gain/loss of function, and Treg functional assays in CD4+ T cell subsets","pmids":["19098714"],"confidence":"High","gaps":["Whether AC9 is the dominant cAMP source in Tregs not established","Direct FOXP3-miR-142-3p promoter interaction details not resolved"]},{"year":2013,"claim":"Extended miRNA control of AC9 to cancer, showing miR-181b suppresses AC9 and that AC9 loss drives proliferation, implicating cAMP output in tumor cell growth.","evidence":"Luciferase reporter, siRNA knockdown and overexpression, cAMP, proliferation and apoptosis assays in cervical cancer cells","pmids":["24269684"],"confidence":"Medium","gaps":["Downstream cAMP effectors of the antiproliferative effect not identified","Single cancer cell context"]},{"year":2014,"claim":"Demonstrated that AC9-derived cAMP enhances retinoic acid receptor activity and ATRA-induced differentiation, with miR-181a (repressed by CEBPalpha) tuning AC9 levels and ATRA sensitivity.","evidence":"3'UTR luciferase reporter, siRNA/overexpression, cAMP, differentiation assays, and CEBPalpha gain/loss of function in APL cells","pmids":["24722286"],"confidence":"Medium","gaps":["Mechanism by which cAMP potentiates RAR trans-activity not detailed","Single lab/cell system"]},{"year":2018,"claim":"Defined the molecular basis of AC9 regulation by identifying the C2b domain (residues 1268-1276) as an intramolecular auto-inhibitor whose removal—possibly by proteolysis—boosts receptor-stimulated cAMP.","evidence":"Deletion and point mutagenesis, stable HEK-293 expression with cAMP assay, and domain-specific immunoblotting of rodent/human myocardial membranes","pmids":["30121334"],"confidence":"High","gaps":["Protease responsible for C2b cleavage not identified","In vivo physiological trigger for de-repression unknown"]},{"year":2018,"claim":"Revealed a CETP-dependent cardiovascular role: Adcy9 inactivation protects against atherosclerosis and improves endothelial function, but only when CETP is absent, establishing ADCY9-CETP genetic epistasis.","evidence":"Adcy9Gt/Gt and CETPtg mice on atherogenic diet with histology, vasorelaxation pharmacology, telemetry and macrophage assays","pmids":["29674325"],"confidence":"High","gaps":["Molecular mechanism connecting ADCY9 cAMP to CETP not defined","Cell type responsible for the protective effect not pinpointed"]},{"year":2020,"claim":"Connected AC9/cAMP to neuronal repair, showing miR-142-3p suppression elevates AC9 to drive cAMP/CREB signaling and axon growth after injury, with AC9 knockdown reversing the benefit.","evidence":"miR-142-3p inhibitor/mimic plus AC9 siRNA in rats, axon length, cAMP/protein quantification and sensory conduction electrophysiology","pmids":["30710569","33416140"],"confidence":"Medium","gaps":["Whether AC9 acts cell-autonomously in neurons not resolved","Relative contribution of AC9 versus other cyclases unclear"]},{"year":2020,"claim":"Implicated ADCY9 in cardiac development, as zebrafish adcy9 knockdown caused cardiac malformation with macrophage infiltration and mmp9 upregulation.","evidence":"Morpholino knockdown in zebrafish with immunofluorescence, RNA-seq and cardiac morphology","pmids":["32321550"],"confidence":"Medium","gaps":["Morpholino off-target effects not excluded by genetic mutant","cAMP-dependence of the phenotype not directly tested"]},{"year":2023,"claim":"Showed Adcy9 inactivation limits infarct size and adverse remodeling after myocardial infarction, again only without CETP, reinforcing ADCY9 as a CETP-conditional cardiovascular target.","evidence":"Adcy9Gt/Gt mice with LAD ligation, echocardiography, histology and immune cell flow cytometry","pmids":["37054880"],"confidence":"Medium","gaps":["Causal cell type for cardioprotection not identified","Link between bone marrow lymphocyte changes and outcome not established"]},{"year":2025,"claim":"Provided variant-level functional evidence in airway disease, showing ADCY9-driven cAMP opposes airway smooth muscle remodeling and that the Ile772Met substitution abolishes this protection.","evidence":"Wild-type versus Ile772Met ADCY9 expression in airway smooth muscle cells with cAMP, remodeling assays and clinical lung-function correlation","pmids":["40816614"],"confidence":"Medium","gaps":["Structural/enzymatic defect caused by Ile772Met not characterized","In vivo airway phenotype of the variant not tested"]},{"year":null,"claim":"The molecular mechanism linking ADCY9 cAMP output to the CETP-dependent cardiovascular phenotype, and the protease/trigger that relieves C2b auto-inhibition in vivo, remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No biochemical pathway connecting ADCY9 and CETP","C2b-cleaving protease unidentified","No structural model of human AC9 regulation"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0009975","term_label":"cyclase activity","supporting_discovery_ids":[0,4]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,4]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,4]}],"complexes":[],"partners":[],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O60503","full_name":"Adenylate cyclase type 9","aliases":["ATP pyrophosphate-lyase 9","Adenylate cyclase type IX","ACIX","Adenylyl cyclase 9","AC9"],"length_aa":1353,"mass_kda":150.7,"function":"Adenylyl cyclase that catalyzes the formation of the signaling molecule cAMP in response to activation of G protein-coupled receptors (PubMed:10987815, PubMed:12972952, PubMed:15879435, PubMed:9628827). Contributes to signaling cascades activated by CRH (corticotropin-releasing factor), corticosteroids and beta-adrenergic receptors (PubMed:9628827)","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/O60503/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ADCY9","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ADCY9","total_profiled":1310},"omim":[{"mim_id":"603302","title":"ADENYLATE CYCLASE 9; ADCY9","url":"https://www.omim.org/entry/603302"},{"mim_id":"155600","title":"MELANOMA, CUTANEOUS MALIGNANT, SUSCEPTIBILITY TO, 1; CMM1","url":"https://www.omim.org/entry/155600"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Plasma membrane","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"},{"location":"Primary cilium","reliability":"Additional"},{"location":"Basal body","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"skeletal muscle","ntpm":48.5}],"url":"https://www.proteinatlas.org/search/ADCY9"},"hgnc":{"alias_symbol":["AC9"],"prev_symbol":[]},"alphafold":{"accession":"O60503","domains":[{"cath_id":"-","chopping":"99-195_218-310","consensus_level":"medium","plddt":86.0725,"start":99,"end":310},{"cath_id":"3.30.70.1230","chopping":"1049-1248_1263-1281","consensus_level":"medium","plddt":86.0799,"start":1049,"end":1281}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O60503","model_url":"https://alphafold.ebi.ac.uk/files/AF-O60503-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O60503-F1-predicted_aligned_error_v6.png","plddt_mean":70.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ADCY9","jax_strain_url":"https://www.jax.org/strain/search?query=ADCY9"},"sequence":{"accession":"O60503","fasta_url":"https://rest.uniprot.org/uniprotkb/O60503.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O60503/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O60503"}},"corpus_meta":[{"pmid":"19098714","id":"PMC_19098714","title":"miR-142-3p restricts cAMP production in CD4+CD25- T cells and CD4+CD25+ TREG cells by targeting AC9 mRNA.","date":"2008","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/19098714","citation_count":227,"is_preprint":false},{"pmid":"9628827","id":"PMC_9628827","title":"Cloning, chromosomal mapping, and regulatory properties of the human type 9 adenylyl cyclase (ADCY9).","date":"1998","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/9628827","citation_count":88,"is_preprint":false},{"pmid":"24269684","id":"PMC_24269684","title":"miR-181b promotes cell proliferation and reduces apoptosis by repressing the expression of adenylyl cyclase 9 (AC9) in cervical cancer cells.","date":"2013","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/24269684","citation_count":62,"is_preprint":false},{"pmid":"21545619","id":"PMC_21545619","title":"Combined pharmacogenetic effect of ADCY9 and ADRB2 gene polymorphisms on the bronchodilator response to inhaled combination therapy.","date":"2010","source":"Journal of clinical pharmacy and therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/21545619","citation_count":44,"is_preprint":false},{"pmid":"29525816","id":"PMC_29525816","title":"ADCY9 Genetic Variants and Cardiovascular Outcomes With Evacetrapib in Patients With High-Risk Vascular Disease: A Nested Case-Control Study.","date":"2018","source":"JAMA cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/29525816","citation_count":38,"is_preprint":false},{"pmid":"29674325","id":"PMC_29674325","title":"ADCY9 (Adenylate Cyclase Type 9) Inactivation Protects From Atherosclerosis Only in the Absence of CETP (Cholesteryl Ester Transfer Protein).","date":"2018","source":"Circulation","url":"https://pubmed.ncbi.nlm.nih.gov/29674325","citation_count":33,"is_preprint":false},{"pmid":"31331193","id":"PMC_31331193","title":"Impact of ADCY9 Genotype on Response to Anacetrapib.","date":"2019","source":"Circulation","url":"https://pubmed.ncbi.nlm.nih.gov/31331193","citation_count":29,"is_preprint":false},{"pmid":"24722286","id":"PMC_24722286","title":"MicroRNA-181a-mediated downregulation of AC9 protein decreases intracellular cAMP level and inhibits ATRA-induced APL cell differentiation.","date":"2014","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/24722286","citation_count":24,"is_preprint":false},{"pmid":"11840511","id":"PMC_11840511","title":"Molecular analysis, mutation screening, and association study of adenylate cyclase type 9 gene (ADCY9) in mood disorders.","date":"2002","source":"American journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/11840511","citation_count":22,"is_preprint":false},{"pmid":"33416140","id":"PMC_33416140","title":"miR‑142‑3p targets AC9 to regulate sciatic nerve injury‑induced neuropathic pain by regulating the cAMP/AMPK signalling pathway.","date":"2020","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33416140","citation_count":21,"is_preprint":false},{"pmid":"35130732","id":"PMC_35130732","title":"Multivalent Interactions Drive the Toxoplasma AC9:AC10:ERK7 Complex To Concentrate ERK7 in the Apical Cap.","date":"2022","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/35130732","citation_count":21,"is_preprint":false},{"pmid":"30121334","id":"PMC_30121334","title":"Auto-inhibition of adenylyl cyclase 9 (AC9) by an isoform-specific motif in the carboxyl-terminal region.","date":"2018","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/30121334","citation_count":20,"is_preprint":false},{"pmid":"30710569","id":"PMC_30710569","title":"Sorafenib promotes sensory conduction function recovery via miR-142-3p/AC9/cAMP axis post dorsal column injury.","date":"2019","source":"Neuropharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/30710569","citation_count":17,"is_preprint":false},{"pmid":"34988676","id":"PMC_34988676","title":"Upregulated microRNA-210-3p improves sevoflurane-induced protective effect on ventricular remodeling in rats with myocardial infarction by inhibiting ADCY9.","date":"2022","source":"Functional & integrative genomics","url":"https://pubmed.ncbi.nlm.nih.gov/34988676","citation_count":10,"is_preprint":false},{"pmid":"34609279","id":"PMC_34609279","title":"A sex-specific evolutionary interaction between ADCY9 and CETP.","date":"2021","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/34609279","citation_count":9,"is_preprint":false},{"pmid":"37065546","id":"PMC_37065546","title":"ADCY9 functions as a novel cancer suppressor gene in lung adenocarcinoma.","date":"2023","source":"Journal of thoracic disease","url":"https://pubmed.ncbi.nlm.nih.gov/37065546","citation_count":6,"is_preprint":false},{"pmid":"32983975","id":"PMC_32983975","title":"A Case-Control Study of ADCY9 Gene Polymorphisms and the Risk of Hepatocellular Carcinoma in the Chinese Han Population.","date":"2020","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/32983975","citation_count":6,"is_preprint":false},{"pmid":"32321550","id":"PMC_32321550","title":"Role of the ADCY9 gene in cardiac abnormalities of the Rubinstein-Taybi syndrome.","date":"2020","source":"Orphanet journal of rare diseases","url":"https://pubmed.ncbi.nlm.nih.gov/32321550","citation_count":6,"is_preprint":false},{"pmid":"34173811","id":"PMC_34173811","title":"Advancing Beyond Failed High-density Lipoprotein Clinical Trials to Pharmacogenetic Studies of ADCY9 and Cholesterol Ester Transfer Protein Inhibition.","date":"2021","source":"Journal of cardiovascular pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/34173811","citation_count":4,"is_preprint":false},{"pmid":"37423551","id":"PMC_37423551","title":"Anti-Inflammatory and Pro-Resolving Actions of the N-Terminal Peptides Ac2-26, Ac2-12, and Ac9-25 of Annexin A1 on Conjunctival Goblet Cell Function.","date":"2023","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/37423551","citation_count":4,"is_preprint":false},{"pmid":"34683946","id":"PMC_34683946","title":"Association between ADCY9 Gene Polymorphisms and Ritodrine Treatment Outcomes in Patients with Preterm Labor.","date":"2021","source":"Pharmaceutics","url":"https://pubmed.ncbi.nlm.nih.gov/34683946","citation_count":3,"is_preprint":false},{"pmid":"39057056","id":"PMC_39057056","title":"ADRB2 and ADCY9 Sequence Variations in Brazilian Asthmatic Patients.","date":"2024","source":"Current issues in molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/39057056","citation_count":3,"is_preprint":false},{"pmid":"37054880","id":"PMC_37054880","title":"Adcy9 Gene Inactivation Improves Cardiac Function After Myocardial Infarction in Mice.","date":"2023","source":"The Canadian journal of cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/37054880","citation_count":1,"is_preprint":false},{"pmid":"37579959","id":"PMC_37579959","title":"The rs2601796 variant in ADCY9 gene is associated with severe asthma and less bronchodilator response.","date":"2023","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/37579959","citation_count":0,"is_preprint":false},{"pmid":"40816614","id":"PMC_40816614","title":"An Ile to Met polymorphism in ADCY9 promotes airway obstruction and remodeling in asthma.","date":"2025","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/40816614","citation_count":0,"is_preprint":false},{"pmid":"36111722","id":"PMC_36111722","title":"[Association of the ADCY9 gene and gene-environmental interaction with the susceptibility to childhood bronchial asthma].","date":"2022","source":"Zhongguo dang dai er ke za zhi = Chinese journal of contemporary pediatrics","url":"https://pubmed.ncbi.nlm.nih.gov/36111722","citation_count":0,"is_preprint":false},{"pmid":"41827823","id":"PMC_41827823","title":"ADCY9 Regulates Neural Stem Cells Via Mitofusin-1 to Maintain Planarian (Dugesia japonica) Cephalic Ganglia Regeneration.","date":"2026","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/41827823","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":17230,"output_tokens":3889,"usd":0.055012,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11621,"output_tokens":3401,"usd":0.071565,"stage2_stop_reason":"end_turn"},"total_usd":0.126577,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1998,\n      \"finding\": \"Human ADCY9 (AC9) is stimulated by beta-adrenergic receptor activation but is insensitive to forskolin, Ca2+, and somatostatin when expressed in HEK-293 cells; unlike mouse AC9, human AC9 activity is unaffected by inhibitors of calcineurin. Divergence at the C2a/C2b junction results in an alternative C2b amino acid sequence compared to mouse AC9.\",\n      \"method\": \"Heterologous expression in HEK-293 cells, pharmacological assays, cAMP measurement\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct functional assay in mammalian cells with multiple pharmacological probes, single lab\",\n      \"pmids\": [\"9628827\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"miR-142-3p targets AC9 mRNA to suppress cAMP production in CD4+CD25- T cells; in CD4+CD25+ T regulatory cells, FOXP3 downregulates miR-142-3p, thereby keeping the AC9/cAMP pathway active and enabling high cAMP levels required for suppressor function.\",\n      \"method\": \"miRNA target validation (luciferase reporter, mRNA/protein quantification), T cell functional assays, cAMP measurement, FOXP3 overexpression/knockdown\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal regulation validated with multiple methods (reporter assay, gain/loss of function, functional T cell assays), replicated across two cell types\",\n      \"pmids\": [\"19098714\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"miR-181b targets AC9 mRNA to restrict cAMP production post-transcriptionally in cervical cancer cells; knockdown of AC9 phenocopies miR-181b overexpression (increased proliferation, reduced apoptosis), while AC9 overexpression has opposite effects.\",\n      \"method\": \"miRNA target validation (luciferase reporter assay), siRNA knockdown and overexpression, cAMP measurement, proliferation and apoptosis assays\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reporter assay plus functional rescue experiments, single lab\",\n      \"pmids\": [\"24269684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"AC9 catalyzes cAMP production in APL cells and enhances retinoic acid receptor trans-activity; AC9 knockdown inhibits ATRA-induced differentiation of NB4 cells. miR-181a targets the 3'UTR of AC9 mRNA to decrease AC9 expression and intracellular cAMP, and CEBPα inhibits miR-181a expression, accounting for differential AC9 levels between ATRA-sensitive and resistant cells.\",\n      \"method\": \"Luciferase reporter assay (3'UTR targeting), siRNA knockdown, overexpression, cAMP measurement, differentiation assays, CEBPα gain/loss of function\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (reporter, KD, OE, functional readout), single lab\",\n      \"pmids\": [\"24722286\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"The isoform-specific carboxyl-terminal C2b domain of AC9 auto-inhibits enzyme activation by Gs-coupled receptors; deletion of the C2b domain markedly enhances cAMP response to beta-adrenergic and prostanoid receptor activation. Residues 1268-1276 within C2b are critical for auto-inhibition. Two molecular weight species of AC9 (~130K and ≥170K) are detected in myocardial membranes, with the lower species lacking the C2b domain, suggesting proteolytic cleavage may regulate AC9 activity.\",\n      \"method\": \"Deletion mutagenesis, stable overexpression in HEK-293 cells, cAMP assay, immunoblotting with domain-specific antibodies, rodent and human myocardial membrane fractionation\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — mutagenesis (deletion and point mutants) with direct enzymatic readout plus tissue biochemistry, single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"30121334\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Adcy9 inactivation in mice reduces aortic atherosclerosis by 65%, decreases macrophage accumulation and proliferation in plaques, and improves endothelial-dependent vasorelaxation via nitric oxide, cyclooxygenase, and endothelial-dependent hyperpolarization pathways. These protective effects are entirely lost when CETP is present (CETPtg Adcy9Gt/Gt mice), demonstrating that Adcy9 inactivation protects from atherosclerosis only in the absence of CETP activity.\",\n      \"method\": \"Genetic inactivation (Adcy9Gt/Gt mice), atherogenic diet protocol, histology (CD68 macrophage staining), vasorelaxation pharmacology, telemetry, adipose tissue MRI, splenocyte adhesion assay\",\n      \"journal\": \"Circulation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — comprehensive in vivo loss-of-function with multiple orthogonal readouts (atherosclerosis, endothelial function, macrophage biology, autonomic tone), epistasis with CETP transgene\",\n      \"pmids\": [\"29674325\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"miR-142-3p targets AC9 in dorsal root ganglion/spinal cord neurons; miR-142-3p inhibition upregulates AC9, elevates cAMP and downstream p-CREB, IL-6, and GAP43, and reduces GTP-RhoA, promoting axon growth in vitro and sensory conduction recovery in vivo after dorsal column injury. Co-knockdown of AC9 reverses these effects. Sorafenib mimics SNCI by downregulating miR-142-3p.\",\n      \"method\": \"miR-142-3p inhibitor/mimic delivery in rats, AC9 siRNA, axon length measurement, cAMP/protein quantification, in vivo sensory conduction electrophysiology\",\n      \"journal\": \"Neuropharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis (miR inhibitor + AC9 siRNA rescue), in vitro and in vivo functional readouts, single lab\",\n      \"pmids\": [\"30710569\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"miR-142-3p directly targets AC9 (validated by luciferase reporter assay) in sciatic nerve injury (CCI) rats; miR-142-3p expression increases while AC9 and cAMP decrease in CCI rats. miR-142-3p silencing reduces neuropathic pain and inflammatory cytokines via upregulation of AC9 and downstream cAMP/AMPK pathway proteins.\",\n      \"method\": \"Dual luciferase reporter assay, miR-142-3p mimic/siRNA delivery in CCI rats, AC9 overexpression/siRNA, cAMP measurement, AMPK pathway protein quantification, pain behavioral testing\",\n      \"journal\": \"International journal of molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — luciferase validation of targeting plus in vivo epistasis experiments, single lab\",\n      \"pmids\": [\"33416140\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Deletion of adcy9 in zebrafish (morphant model) causes cardiac malformation with increased macrophage migration, cardiac apoptosis, and upregulation of mmp9 (matrix metalloproteinase involved in extracellular matrix remodeling), supporting ADCY9 as a candidate gene for congenital heart defects.\",\n      \"method\": \"Zebrafish adcy9 morpholino knockdown, immunofluorescence (macrophage markers), RNA sequencing, cardiac morphology assessment\",\n      \"journal\": \"Orphanet journal of rare diseases\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo loss-of-function with immunofluorescence and transcriptomics, single lab, zebrafish model\",\n      \"pmids\": [\"32321550\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"miR-210-3p targets ADCY9 mRNA (validated by target relation detection); miR-210-3p is downregulated while ADCY9 is upregulated in myocardial infarction rats. Sevoflurane-mediated cardioprotection (reduced injury, fibrosis, and apoptosis) is potentiated by miR-210-3p elevation and abolished by miR-210-3p inhibition or ADCY9 overexpression, placing ADCY9 downstream of miR-210-3p in the sevoflurane cardioprotective pathway.\",\n      \"method\": \"miR-210-3p agomir/antagomir delivery, ADCY9 overexpression in MI rat model, echocardiography, histology, miRNA-target validation assay\",\n      \"journal\": \"Functional & integrative genomics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — functional rescue experiments in vivo but target validation method not fully described in abstract, single lab\",\n      \"pmids\": [\"34988676\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Adcy9 inactivation in mice after myocardial infarction reduces infarct size, pathological left ventricular remodeling, and cardiac dysfunction, and preserves myocardial capillary density in the infarct border zone; increased bone marrow T cells and B cells were also observed. These benefits were only seen in the absence of CETP activity.\",\n      \"method\": \"Adcy9Gt/Gt mice, permanent LAD ligation MI model, echocardiography, histology (cardiomyocyte size, infarct size, capillary density), flow cytometry (immune cells)\",\n      \"journal\": \"The Canadian journal of cardiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic loss-of-function with multiple histological and functional readouts, CETP epistasis, single lab\",\n      \"pmids\": [\"37054880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ADCY9 overexpression in lung adenocarcinoma cell lines (SPCA1, A549) restrains cell proliferation, invasion, and migration, consistent with a tumor-suppressive function mediated through the cAMP pathway.\",\n      \"method\": \"ADCY9 overexpression in cancer cell lines, proliferation assay, invasion/migration assays\",\n      \"journal\": \"Journal of thoracic disease\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — overexpression phenotype assays only, no direct mechanistic pathway validation, single lab\",\n      \"pmids\": [\"37065546\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Asthma-related cytokines decrease ADCY9 expression and cellular cAMP levels in airway smooth muscle cells (ASMCs), reducing airway relaxation and promoting remodeling. ADCY9 overexpression attenuates airway smooth muscle remodeling, but the missense variant ADCY9-Ile772Met (rs2230739) fails to prevent remodeling, demonstrating that the Ile772Met substitution impairs ADCY9's protective function in the airway.\",\n      \"method\": \"ADCY9 overexpression and mutant (Ile772Met) expression in ASMCs, cAMP measurement, airway remodeling assays, FEV1/FVC% clinical correlation\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional comparison of wild-type vs missense mutant with enzymatic (cAMP) and cellular readouts, single lab\",\n      \"pmids\": [\"40816614\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2027,\n      \"finding\": \"In planarian (Dugesia japonica), ADCY9 knockdown causes abnormal brain regeneration (loss of nerve cord, reduced collateral branches, inhibited neuron regeneration/differentiation). Double RNAi of ADCY9 and Mitofusin-1 restores neural regeneration, indicating ADCY9 promotes neural regeneration by negatively regulating the downstream inhibitory factor Mitofusin-1 in the cAMP/AMPK signaling pathway.\",\n      \"method\": \"RNAi knockdown (single and double) in planarians, RNA sequencing, KEGG pathway analysis, morphological assessment of brain regeneration\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — genetic epistasis by double RNAi in planarian model, single lab, no direct biochemical validation of ADCY9-Mitofusin-1 interaction\",\n      \"pmids\": [\"41827823\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ADCY9 (AC9) is a transmembrane adenylyl cyclase that catalyzes ATP-to-cAMP conversion and is activated by Gs-coupled receptors (including beta-adrenergic receptors) but is insensitive to forskolin and Ca2+; its activity is auto-inhibited by an isoform-specific C2b carboxyl-terminal domain (residues 1268-1276), potentially relieved by proteolytic cleavage; ADCY9 is post-transcriptionally suppressed by multiple miRNAs (miR-142-3p, miR-181a, miR-181b, miR-210-3p) that target its 3'UTR to reduce intracellular cAMP, with FOXP3 counter-regulating miR-142-3p in T regulatory cells to maintain high cAMP; in vivo, ADCY9 inactivation protects against atherosclerosis, reduces myocardial infarction injury, and improves endothelial function exclusively in the absence of CETP activity, revealing a genetic epistatic interaction between ADCY9 and CETP.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ADCY9 (AC9) is a transmembrane adenylyl cyclase that synthesizes cAMP downstream of Gs-coupled receptors, including beta-adrenergic and prostanoid receptors, and is distinguished from other adenylyl cyclases by its insensitivity to forskolin and Ca2+ [#0]. Its activation is restrained by an isoform-specific carboxyl-terminal C2b domain (residues 1268-1276), whose deletion markedly amplifies the cAMP response to receptor stimulation; two molecular-weight AC9 species in myocardial membranes indicate that proteolytic removal of C2b can relieve this auto-inhibition [#4]. ADCY9 output is set post-transcriptionally by multiple miRNAs that target its 3'UTR—miR-142-3p, miR-181a/b, and miR-210-3p—each lowering AC9 protein and intracellular cAMP, with upstream regulators such as FOXP3 and CEBPalpha tuning these miRNAs to control cAMP in T regulatory cells and during myeloid differentiation [#1, #3]. Through this cAMP-generating activity, ADCY9 governs diverse physiological outputs: it sustains regulatory T cell suppressor function [#1], supports retinoic-acid-induced differentiation [#3], and shapes cardiovascular biology, where genetic Adcy9 inactivation reduces atherosclerosis, limits myocardial infarction injury, and improves endothelial function—effects observed exclusively in the absence of CETP activity, defining a genetic epistasis between ADCY9 and CETP [#5, #10]. In airway smooth muscle, ADCY9-driven cAMP promotes relaxation and opposes remodeling, a protective function abolished by the missense variant Ile772Met [#12].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Established that human AC9 is a functional adenylyl cyclase coupled to Gs receptors yet pharmacologically atypical, defining its identity among the cyclase family.\",\n      \"evidence\": \"Heterologous expression in HEK-293 cells with beta-adrenergic, forskolin, Ca2+ and calcineurin probes plus cAMP measurement\",\n      \"pmids\": [\"9628827\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural basis for forskolin insensitivity defined\", \"Endogenous receptor partners in native tissue not mapped\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Showed that AC9 is post-transcriptionally controlled by miR-142-3p, and that FOXP3 relieves this repression to maintain the high cAMP required for Treg suppressor function, linking AC9 to immune regulation.\",\n      \"evidence\": \"Luciferase reporter, mRNA/protein quantification, FOXP3 gain/loss of function, and Treg functional assays in CD4+ T cell subsets\",\n      \"pmids\": [\"19098714\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether AC9 is the dominant cAMP source in Tregs not established\", \"Direct FOXP3-miR-142-3p promoter interaction details not resolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Extended miRNA control of AC9 to cancer, showing miR-181b suppresses AC9 and that AC9 loss drives proliferation, implicating cAMP output in tumor cell growth.\",\n      \"evidence\": \"Luciferase reporter, siRNA knockdown and overexpression, cAMP, proliferation and apoptosis assays in cervical cancer cells\",\n      \"pmids\": [\"24269684\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Downstream cAMP effectors of the antiproliferative effect not identified\", \"Single cancer cell context\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated that AC9-derived cAMP enhances retinoic acid receptor activity and ATRA-induced differentiation, with miR-181a (repressed by CEBPalpha) tuning AC9 levels and ATRA sensitivity.\",\n      \"evidence\": \"3'UTR luciferase reporter, siRNA/overexpression, cAMP, differentiation assays, and CEBPalpha gain/loss of function in APL cells\",\n      \"pmids\": [\"24722286\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which cAMP potentiates RAR trans-activity not detailed\", \"Single lab/cell system\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined the molecular basis of AC9 regulation by identifying the C2b domain (residues 1268-1276) as an intramolecular auto-inhibitor whose removal—possibly by proteolysis—boosts receptor-stimulated cAMP.\",\n      \"evidence\": \"Deletion and point mutagenesis, stable HEK-293 expression with cAMP assay, and domain-specific immunoblotting of rodent/human myocardial membranes\",\n      \"pmids\": [\"30121334\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Protease responsible for C2b cleavage not identified\", \"In vivo physiological trigger for de-repression unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Revealed a CETP-dependent cardiovascular role: Adcy9 inactivation protects against atherosclerosis and improves endothelial function, but only when CETP is absent, establishing ADCY9-CETP genetic epistasis.\",\n      \"evidence\": \"Adcy9Gt/Gt and CETPtg mice on atherogenic diet with histology, vasorelaxation pharmacology, telemetry and macrophage assays\",\n      \"pmids\": [\"29674325\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism connecting ADCY9 cAMP to CETP not defined\", \"Cell type responsible for the protective effect not pinpointed\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected AC9/cAMP to neuronal repair, showing miR-142-3p suppression elevates AC9 to drive cAMP/CREB signaling and axon growth after injury, with AC9 knockdown reversing the benefit.\",\n      \"evidence\": \"miR-142-3p inhibitor/mimic plus AC9 siRNA in rats, axon length, cAMP/protein quantification and sensory conduction electrophysiology\",\n      \"pmids\": [\"30710569\", \"33416140\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether AC9 acts cell-autonomously in neurons not resolved\", \"Relative contribution of AC9 versus other cyclases unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Implicated ADCY9 in cardiac development, as zebrafish adcy9 knockdown caused cardiac malformation with macrophage infiltration and mmp9 upregulation.\",\n      \"evidence\": \"Morpholino knockdown in zebrafish with immunofluorescence, RNA-seq and cardiac morphology\",\n      \"pmids\": [\"32321550\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Morpholino off-target effects not excluded by genetic mutant\", \"cAMP-dependence of the phenotype not directly tested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed Adcy9 inactivation limits infarct size and adverse remodeling after myocardial infarction, again only without CETP, reinforcing ADCY9 as a CETP-conditional cardiovascular target.\",\n      \"evidence\": \"Adcy9Gt/Gt mice with LAD ligation, echocardiography, histology and immune cell flow cytometry\",\n      \"pmids\": [\"37054880\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Causal cell type for cardioprotection not identified\", \"Link between bone marrow lymphocyte changes and outcome not established\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Provided variant-level functional evidence in airway disease, showing ADCY9-driven cAMP opposes airway smooth muscle remodeling and that the Ile772Met substitution abolishes this protection.\",\n      \"evidence\": \"Wild-type versus Ile772Met ADCY9 expression in airway smooth muscle cells with cAMP, remodeling assays and clinical lung-function correlation\",\n      \"pmids\": [\"40816614\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural/enzymatic defect caused by Ile772Met not characterized\", \"In vivo airway phenotype of the variant not tested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The molecular mechanism linking ADCY9 cAMP output to the CETP-dependent cardiovascular phenotype, and the protease/trigger that relieves C2b auto-inhibition in vivo, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No biochemical pathway connecting ADCY9 and CETP\", \"C2b-cleaving protease unidentified\", \"No structural model of human AC9 regulation\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0009975\", \"supporting_discovery_ids\": [0, 4]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 4]}\n    ],\n    \"complexes\": [],\n    \"partners\": [],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}