{"gene":"ADCY3","run_date":"2026-06-09T22:02:41","timeline":{"discoveries":[{"year":2018,"finding":"ADCY3 localizes to the primary cilia of neurons, and MC4R colocalizes with ADCY3 at the primary cilia of a subset of hypothalamic neurons. Obesity-associated MC4R mutations impair ciliary localization of MC4R, and inhibition of adenylyl cyclase signaling at the primary cilia of these neurons increases body weight, placing ADCY3-dependent cAMP signaling at neuronal primary cilia as a common pathway underlying genetic causes of obesity.","method":"Confocal colocalization imaging in hypothalamic neurons, pharmacological inhibition of adenylyl cyclase in vivo, genetic mouse models with obesity-associated MC4R mutations","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (colocalization, in vivo pharmacology, genetic mutation analysis), replicated context across three concurrent papers in same issue","pmids":["29311635"],"is_preprint":false},{"year":2018,"finding":"Homozygous and compound heterozygous loss-of-function mutations in ADCY3 cause severe monogenic obesity in humans, establishing ADCY3 as a mediator of energy homeostasis. Functional characterization of these mutations confirmed loss of adenylate cyclase 3 activity.","method":"Human genetic sequencing, functional characterization of mutations in patient-derived samples","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — functional characterization of mutations combined with human genetics, replicated across independent consanguineous families and different ethnic backgrounds","pmids":["29311637"],"is_preprint":false},{"year":2018,"finding":"A splice-acceptor-site variant in ADCY3 decreases ADCY3 RNA expression and is associated with markedly increased risk of obesity and type 2 diabetes, confirming ADCY3 loss-of-function as pathogenic for metabolic disease.","method":"Population genetics, splice-site variant identification, RNA expression analysis in carriers","journal":"Nature genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNA expression measured in carriers, single population (Greenlandic), replicated direction in trans-ancestry cohorts","pmids":["29311636"],"is_preprint":false},{"year":2013,"finding":"ADCY3 overexpression increases cAMP levels and activates the cAMP/PKA/CREB pathway, leading to increased MMP2 and MMP9 mRNA expression and activity. PKA inhibitor treatment decreased MMP2 and MMP9 expression in ADCY3-overexpressing cells, placing ADCY3 upstream of PKA and CREB in this signaling cascade. ADCY3 expression is regulated by promoter CpG methylation.","method":"Overexpression and shRNA knockdown in HEK293 and SNU-216 cells, PKA inhibitor treatment, cAMP measurement, luciferase reporter assay, tumor xenograft model, bisulfite sequencing","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (cAMP measurement, PKA inhibition, gene knockdown, in vivo xenograft), single lab","pmids":["24113161"],"is_preprint":false},{"year":2012,"finding":"In Drosophila M pacemaker neurons, adenylate cyclase AC3 (ortholog of mammalian ADCY3) specifically mediates PDF neuropeptide receptor (PDF-R) signaling to produce cAMP increases. Genetic disruption of AC3 selectively abolishes PDF-induced cAMP responses in M cells without affecting other Gs-coupled GPCR signaling. The AKAP-like scaffolding protein Nervy also reduces PDF responses when knocked down, suggesting PDF-R/AC3 are organized into a 'circadian signalosome'. AC3 is not required for PDF signaling in E pacemaker cells, demonstrating cell-type-specific coupling.","method":"Live imaging of intact fly brains with cAMP reporters, transgenic RNAi knockdown, genetic rescue experiments, behavioral analysis","journal":"PLoS biology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — live in vivo cAMP imaging combined with genetic RNAi and rescue, multiple cell-type comparisons, rigorous controls excluding developmental effects","pmids":["22679392"],"is_preprint":false},{"year":2024,"finding":"ADCY3 localizes dynamically to neuronal primary cilia in the mouse brain in a region- and age-dependent manner. ARL13B+ cilia become relatively rare with age in hypothalamic feeding centers, while ADCY3 becomes a prominent cilia marker in the mature adult brain, indicating dynamic changes in cilia protein composition during postnatal development.","method":"Immunofluorescence imaging of mouse brain sections across postnatal ages and brain regions, quantification of cilia marker proportions","journal":"Cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiment with region- and age-dependent quantification, multiple brain regions and time points, single lab","pmids":["38334651"],"is_preprint":false},{"year":2024,"finding":"A protein-coding mutation in the transmembrane (TM) domain of Adcy3 in rats (Adcy3mut/mut) increases adiposity and alters emotional behaviors in a sex-dependent manner. Male Adcy3mut/mut rats showed increased passive coping and decreased memory, while females showed increased anxiety-like behavior. Adcy3mut/mut males had decreased hypothalamic CREB signaling, with decreased p-AMPK signaling in both sexes, linking the TM domain of ADCY3 to cAMP/CREB and AMPK signaling.","method":"CRISPR-SpCas9 TM-domain mutation in WKY rats, high-fat diet feeding, behavioral tests, Western blotting for CREB and p-AMPK","journal":"Obesity (Silver Spring, Md.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean CRISPR KO and coding mutation model, multiple behavioral and biochemical readouts, single lab","pmids":["39632398"],"is_preprint":false},{"year":2023,"finding":"The Akkermansia muciniphila membrane protein Amuc_1100 promotes lipolysis and browning in 3T3-L1 preadipocytes by upregulating the AC3/PKA/HSL pathway, increasing lipolysis-related genes (AC3, PKA, HSL) and phosphorylating HSL at Ser660, placing ADCY3 upstream of PKA and HSL in adipocyte lipolysis signaling.","method":"Transcriptomics in 3T3-L1 preadipocytes, qPCR, Western blotting, in vivo and in vitro fat accumulation assays","journal":"Microbiology spectrum","confidence":"Low","confidence_rationale":"Tier 3 / Weak — Western blotting and gene expression data only, indirect activation of ADCY3 by a bacterial protein, single lab","pmids":["36847500"],"is_preprint":false},{"year":2019,"finding":"Liraglutide upregulates hepatic AC3 expression and cAMP/PKA activity in obese mice, leading to increased phosphorylated HSL (p-HSL Ser660) and promoting lipolysis via the AC3/PKA/HSL pathway.","method":"In vivo liraglutide treatment of obese mice, Western blotting, RT-qPCR, cAMP and PKA activity assays in liver tissue","journal":"Diabetes, metabolic syndrome and obesity : targets and therapy","confidence":"Low","confidence_rationale":"Tier 3 / Weak — in vivo pharmacological upregulation study, indirect activation of ADCY3, single lab, no direct mechanistic perturbation of ADCY3","pmids":["31564937"],"is_preprint":false},{"year":2024,"finding":"In vivo experiments in a rat epilepsy model show that H2S targets and downregulates AC3 expression, thereby modulating the AC3/cAMP/PKA signaling pathway to regulate TRPV2 expression. The AC3 activator forskolin reversed the downregulatory effects of H2S on this pathway, establishing AC3 as a functional node upstream of cAMP/PKA/TRPV2 in seizure regulation.","method":"Proteomics in LiCl/Pilocarpine-induced seizure rat model, Western blotting, immunofluorescence, pharmacological rescue with AC3 activator forskolin, EEG and behavioral tests","journal":"Neurochemistry international","confidence":"Low","confidence_rationale":"Tier 3 / Weak — pharmacological rescue with forskolin supports pathway placement, but ADCY3 is identified as a target by proteomics without direct mechanistic perturbation, single lab","pmids":["38290616"],"is_preprint":false},{"year":2024,"finding":"A novel homozygous nonsense variant in ADCY3 (p.Thr840X) causes severe early-onset obesity. In vitro and in silico functional analyses showed downregulation and impaired activation of the ADCY3 protein, confirming loss of enzymatic activity as the pathogenic mechanism.","method":"Gene panel sequencing, in vitro functional assay of ADCY3 activity, in silico protein modeling","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 1–2 / Weak — in vitro enzymatic activity assay combined with in silico modeling, single lab, single patient","pmids":["39519366"],"is_preprint":false},{"year":2025,"finding":"ADCY3 catalyzes the production of cAMP in adipose tissue and is rhythmically expressed there with BMAL1 binding near the Ser107/Pro107 site. A transmembrane domain mutation in Adcy3 causes partial loss of enzymatic function, decreasing cAMP production in response to β-3 adrenergic receptor agonist stimulation in adipose tissue ex vivo, reducing serum free fatty acids and adipose triglyceride lipase expression, and contributing to increased adiposity via decreased lipolytic responsiveness.","method":"CRISPR rat model, ex vivo adipose tissue cAMP production assay with β-3 adrenergic receptor agonist, serum FFA measurement, gene expression analysis, cold exposure body temperature measurement","journal":"Research square (preprint)","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — direct in vitro enzymatic function assay (ex vivo cAMP) combined with genetic loss-of-function rat model and multiple metabolic readouts, single lab, preprint","pmids":["41542040"],"is_preprint":true},{"year":2025,"finding":"In mouse olfactory bulb astrocytes, AC3 (along with AC1) acts downstream of α1 adrenergic receptors as a Ca2+/calmodulin-dependent adenylyl cyclase to produce cAMP in response to norepinephrine/phenylephrine, linking Ca2+ signaling to cAMP production through ADCY3.","method":"Live imaging with cAMP and Ca2+ reporters in mouse olfactory bulb astrocytes, pharmacological receptor subtype dissection, Ca2+ depletion experiments","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live imaging with reporters and pharmacological dissection establish pathway order, multiple receptor subtypes tested, single lab, preprint","pmids":["bio_10.1101_2025.11.10.687535"],"is_preprint":true},{"year":2024,"finding":"In a mouse carotid body model of chronic intermittent hypoxia (CIH), Adcy3-dependent cAMP production is downstream of Olfr78 activation by H2S. CIH increases cAMP in the carotid body, absent in Adcy3, Cth, and Olfr78 null mice. CIH-induced cAMP via ADCY3 mediates enhanced Ca2+ influx through cyclic nucleotide-gated channels (CNGA2). Adcy3 null mice do not exhibit carotid body activation or autonomic dysfunction in response to CIH.","method":"Adcy3 null mouse model, cAMP measurement in carotid body, Ca2+ imaging in glomus cells, comparison across Adcy3, Olfr78, Cth, and Cnga2 mutant mice","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple genetic null models with biochemical and cellular readouts establish pathway position, single lab, preprint","pmids":["bio_10.1101_2024.09.24.614747"],"is_preprint":true},{"year":2026,"finding":"The obesity risk SNP rs713586-C allele reduces ADCY3 promoter activity via impaired ZFP42 transcription factor binding, leading to decreased TET1 recruitment and increased DNA methylation at the ADCY3 enhancer and promoter regions, suppressing ADCY3 expression. Dnajc27 knockout mice did not develop obesity, excluding DNAJC27 as the functional target of rs713586, while ADCY3 downregulation was confirmed as the relevant mechanism.","method":"Dual-luciferase reporter assay, CRISPR/Cas9 genome editing in cell lines, Dnajc27 knockout mice, bisulfite sequencing for DNA methylation, ZFP42-TET1 interaction analysis","journal":"EBioMedicine","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (reporter assay, genome editing, KO mice, DNA methylation, TF binding), single lab but comprehensive mechanistic dissection","pmids":["41494241"],"is_preprint":false},{"year":2025,"finding":"A transmembrane domain mutation in Adcy3 decreases hypothalamic cAMP production in rats without altering ADCY3 membrane content, demonstrating that the TM domain is required for enzymatic (cAMP-generating) function. Adcy3mut/mut rats showed sex-specific depression- and anxiety-like behaviors and food seeking, with increased leptin levels in males.","method":"CRISPR rat model, hypothalamic cAMP measurement, membrane fractionation and Western blotting, comprehensive behavioral testing","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct cAMP measurement in hypothalamus combined with membrane fractionation and behavioral battery, single lab, preprint","pmids":["bio_10.1101_2025.03.28.645767"],"is_preprint":true},{"year":2025,"finding":"ADCY3 overexpression in adipocytes impairs adipogenesis by downregulating the adipogenic transcription factors CEBPα and PPARγ, reduces lipid droplet number and size by suppressing triglyceride synthesis and fatty acid metabolism genes (DGAT1, DGAT2, ACC, SCD, FASN, ACSL1), and suppresses oxidative phosphorylation genes through PPARγ signaling.","method":"ADCY3 overexpression in adipocytes, transcriptomic profiling, gene expression and protein analysis (qPCR, Western blotting), lipid droplet quantification","journal":"Functional & integrative genomics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — overexpression study with transcriptomics and gene expression, single lab, no direct measurement of cAMP or ADCY3 enzymatic activity","pmids":["41350952"],"is_preprint":false}],"current_model":"ADCY3 is a transmembrane adenylyl cyclase that localizes to neuronal primary cilia (especially in hypothalamic feeding centers) and catalyzes conversion of ATP to cAMP; it acts downstream of Gs-coupled GPCRs (including MC4R and olfactory receptor Olfr78) and Ca2+/calmodulin signaling to generate cAMP, which activates PKA/CREB and PKA/HSL signaling to regulate energy homeostasis, lipolysis, and behavior, with loss-of-function causing severe obesity in humans and rodents through impaired cAMP-mediated signaling in hypothalamic cilia, adipose tissue, and other metabolically relevant tissues."},"narrative":{"mechanistic_narrative":"ADCY3 is a transmembrane adenylyl cyclase that catalyzes ATP-to-cAMP conversion at neuronal primary cilia and in metabolic tissues, functioning as a central node linking Gs-coupled GPCR and Ca2+ signaling to cAMP-dependent control of energy homeostasis and behavior [PMID:29311635, PMID:29311637]. In the hypothalamus, ADCY3 localizes to the primary cilia of feeding-center neurons where it colocalizes with MC4R, and disruption of ciliary adenylyl cyclase signaling increases body weight, defining ADCY3-dependent ciliary cAMP as a convergence point for genetic obesity [PMID:29311635]; its enzymatic function requires the transmembrane domain, as a TM-domain mutation reduces hypothalamic cAMP production without altering membrane content [PMID:bio_10.1101_2025.03.28.645767]. Loss-of-function in humans causes severe monogenic obesity: homozygous and compound heterozygous mutations, a nonsense variant (p.Thr840X), and regulatory variants that reduce ADCY3 expression all abolish or diminish adenylyl cyclase activity and confer obesity and type 2 diabetes risk [PMID:29311637, PMID:29311636, PMID:39519366]. The obesity risk allele rs713586-C suppresses ADCY3 transcription through impaired ZFP42 binding, reduced TET1 recruitment, and increased promoter/enhancer DNA methylation, with DNAJC27 excluded as the functional target [PMID:41494241]. Downstream, ADCY3-generated cAMP activates PKA to drive CREB-dependent transcription and HSL (Ser660)-dependent lipolysis in adipose and hepatic tissue, where ADCY3 is rhythmically expressed and required for beta-3 adrenergic lipolytic responsiveness [PMID:24113161, PMID:41542040]. ADCY3 also operates as a Ca2+/calmodulin-dependent cyclase downstream of alpha1 adrenergic receptors in astrocytes [PMID:bio_10.1101_2025.11.10.687535] and, in Drosophila pacemaker neurons, mediates cell-type-specific PDF receptor signaling within a putative circadian signalosome [PMID:22679392].","teleology":[{"year":2012,"claim":"Established that ADCY3 is not a generic cAMP source but couples selectively to specific GPCRs in a cell-type-restricted manner, introducing the concept of a localized signalosome.","evidence":"Live cAMP imaging with transgenic RNAi and rescue in Drosophila M and E pacemaker neurons","pmids":["22679392"],"confidence":"High","gaps":["Done in fly ortholog; mammalian cell-type selectivity not directly demonstrated here","Molecular basis of receptor-cyclase coupling specificity unresolved","Role of Nervy/AKAP scaffold in mammals untested"]},{"year":2013,"claim":"Placed ADCY3 upstream of the cAMP/PKA/CREB cascade as a driver of downstream transcriptional output, and showed its expression is epigenetically controlled.","evidence":"Overexpression/knockdown, PKA inhibition, cAMP and luciferase assays, xenograft, and bisulfite sequencing in HEK293 and gastric cancer cells","pmids":["24113161"],"confidence":"Medium","gaps":["Cancer cell context, not metabolic tissue","MMP induction is downstream and indirect","Methylation regulation not linked to physiological signal"]},{"year":2018,"claim":"Resolved the cellular site and physiological role of ADCY3 in obesity by localizing it to hypothalamic neuronal primary cilia alongside MC4R, defining ciliary cAMP as a common obesity pathway.","evidence":"Confocal colocalization, in vivo adenylyl cyclase inhibition, and genetic mouse models of MC4R mutations","pmids":["29311635"],"confidence":"High","gaps":["Direct demonstration that MC4R signals through ADCY3 enzymatically not shown","Identity of full ciliary receptor repertoire incomplete","Downstream neuronal effectors of ciliary cAMP not defined"]},{"year":2018,"claim":"Established ADCY3 loss-of-function as causal for human monogenic obesity, converting a candidate into a validated disease gene.","evidence":"Human sequencing across consanguineous families plus functional characterization confirming loss of cyclase activity; independent population genetics on a splice variant reducing RNA expression","pmids":["29311637","29311636"],"confidence":"High","gaps":["Splice-variant finding limited to single population (Medium confidence)","Tissue-specific contribution of each loss-of-function mechanism not dissected","Genotype-phenotype dose relationship incomplete"]},{"year":2024,"claim":"Localized the enzymatic function to the transmembrane domain and showed ADCY3 ciliary occupancy changes dynamically with brain region and age, refining how and where cAMP is generated.","evidence":"CRISPR TM-domain mutant rats with hypothalamic cAMP measurement and membrane fractionation; immunofluorescence of cilia markers across mouse brain ages","pmids":["39632398","bio_10.1101_2025.03.28.645767","38334651"],"confidence":"Medium","gaps":["Structural basis of TM-domain requirement for catalysis unresolved","Functional consequence of age-dependent ciliary remodeling untested","Sex-specific behavioral mechanisms unexplained"]},{"year":2024,"claim":"Positioned ADCY3 downstream of olfactory receptor Olfr78 and of Ca2+/calmodulin signaling, extending its role beyond MC4R to chemosensory and Ca2+-coupled cAMP generation.","evidence":"Adcy3 null and Olfr78/Cth/Cnga2 mutant mice in carotid body CIH model; live cAMP/Ca2+ imaging with receptor dissection in olfactory bulb astrocytes","pmids":["bio_10.1101_2024.09.24.614747","bio_10.1101_2025.11.10.687535"],"confidence":"Medium","gaps":["Both are preprints","Direct physical coupling of Olfr78/alpha1-AR to ADCY3 not shown","Generalizability beyond carotid body and astrocytes unknown"]},{"year":2025,"claim":"Defined ADCY3's peripheral metabolic role by showing it drives beta-3 adrenergic, rhythmically regulated cAMP/PKA/HSL lipolysis in adipose and liver, with overexpression conversely impairing adipogenesis.","evidence":"Ex vivo adipose cAMP assay in CRISPR rats with BMAL1-binding analysis; liraglutide and Amuc_1100 pathway studies; ADCY3-overexpression adipocyte transcriptomics","pmids":["41542040","31564937","36847500","41350952"],"confidence":"Medium","gaps":["Adipose mechanism partly preprint and partly Low-confidence indirect activation studies","Direct cAMP/enzymatic measurement absent in several adipocyte studies","Reconciliation of overexpression vs loss-of-function phenotypes incomplete"]},{"year":2026,"claim":"Mechanistically explained a major obesity GWAS signal by tracing the rs713586-C allele to ZFP42/TET1-dependent DNA methylation that suppresses ADCY3 expression, and excluded the neighboring DNAJC27.","evidence":"Dual-luciferase assays, CRISPR editing, Dnajc27 knockout mice, bisulfite sequencing, and ZFP42-TET1 interaction analysis","pmids":["41494241"],"confidence":"High","gaps":["In vivo demonstration of allele-specific methylation in human tissue limited","Tissue specificity of the regulatory effect not fully mapped","Single lab"]},{"year":null,"claim":"How ADCY3 achieves receptor- and tissue-specific coupling, and the structural basis by which the transmembrane domain governs catalysis, remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structure-function model linking TM domain to active site","Mechanism of selective GPCR-ADCY3 pairing across cell types unknown","Direct physical interactome of ADCY3 not defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0009975","term_label":"cyclase activity","supporting_discovery_ids":[1,3,10,11,15]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[3,11,15]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,5]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[15]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,4,12,13]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[1,11]}],"complexes":[],"partners":["MC4R","OLFR78"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O60266","full_name":"Adenylate cyclase type 3","aliases":["ATP pyrophosphate-lyase 3","Adenylate cyclase type III","AC-III","Adenylate cyclase, olfactive type","Adenylyl cyclase 3","AC3"],"length_aa":1144,"mass_kda":129.0,"function":"Catalyzes the formation of the signaling molecule cAMP in response to G-protein signaling. Participates in signaling cascades triggered by odorant receptors via its function in cAMP biosynthesis: specifically activated by G alpha protein GNAL/G(olf) in olfactory epithelium. Required for normal sperm motility and normal male fertility. Plays a role in regulating insulin levels and body fat accumulation in response to a high fat diet","subcellular_location":"Cell membrane; Cytoplasm; Cell projection, cilium; Golgi apparatus","url":"https://www.uniprot.org/uniprotkb/O60266/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ADCY3","classification":"Not Classified","n_dependent_lines":55,"n_total_lines":1208,"dependency_fraction":0.04552980132450331},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ADCY3","total_profiled":1310},"omim":[{"mim_id":"618696","title":"GOLGI-ASSOCIATED OLFACTORY SIGNALING REGULATOR; GFY","url":"https://www.omim.org/entry/618696"},{"mim_id":"617885","title":"BODY MASS INDEX QUANTITATIVE TRAIT LOCUS 19; BMIQ19","url":"https://www.omim.org/entry/617885"},{"mim_id":"616629","title":"SENIOR-LOKEN SYNDROME 9; SLSN9","url":"https://www.omim.org/entry/616629"},{"mim_id":"615636","title":"JOUBERT SYNDROME 21; JBTS21","url":"https://www.omim.org/entry/615636"},{"mim_id":"614144","title":"B9 DOMAIN-CONTAINING PROTEIN 1; B9D1","url":"https://www.omim.org/entry/614144"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Uncertain","locations":[{"location":"Nucleoplasm","reliability":"Uncertain"},{"location":"Primary cilium","reliability":"Uncertain"},{"location":"Nucleoli","reliability":"Additional"},{"location":"Golgi apparatus","reliability":"Additional"},{"location":"Plasma membrane","reliability":"Additional"},{"location":"Primary cilium transition zone","reliability":"Additional"},{"location":"Basal body","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ADCY3"},"hgnc":{"alias_symbol":["AC3"],"prev_symbol":[]},"alphafold":{"accession":"O60266","domains":[{"cath_id":"3.30.70.1230","chopping":"310-496","consensus_level":"medium","plddt":88.9897,"start":310,"end":496},{"cath_id":"3.30.70.1230","chopping":"912-990_1006-1126","consensus_level":"medium","plddt":88.7901,"start":912,"end":1126}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O60266","model_url":"https://alphafold.ebi.ac.uk/files/AF-O60266-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O60266-F1-predicted_aligned_error_v6.png","plddt_mean":76.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ADCY3","jax_strain_url":"https://www.jax.org/strain/search?query=ADCY3"},"sequence":{"accession":"O60266","fasta_url":"https://rest.uniprot.org/uniprotkb/O60266.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O60266/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O60266"}},"corpus_meta":[{"pmid":"29311635","id":"PMC_29311635","title":"Subcellular localization of MC4R with ADCY3 at neuronal primary cilia underlies a common pathway for genetic predisposition to obesity.","date":"2018","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29311635","citation_count":205,"is_preprint":false},{"pmid":"29311636","id":"PMC_29311636","title":"Loss-of-function variants in ADCY3 increase risk of obesity and type 2 diabetes.","date":"2018","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29311636","citation_count":147,"is_preprint":false},{"pmid":"29311637","id":"PMC_29311637","title":"Loss-of-function mutations in ADCY3 cause monogenic severe obesity.","date":"2018","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29311637","citation_count":139,"is_preprint":false},{"pmid":"25044758","id":"PMC_25044758","title":"Genome-wide association study of height-adjusted BMI in childhood identifies functional variant in ADCY3.","date":"2014","source":"Obesity (Silver Spring, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/25044758","citation_count":81,"is_preprint":false},{"pmid":"1585657","id":"PMC_1585657","title":"Regulation of the activities of African cassava mosaic virus promoters by the AC1, AC2, and AC3 gene products.","date":"1992","source":"Virology","url":"https://pubmed.ncbi.nlm.nih.gov/1585657","citation_count":80,"is_preprint":false},{"pmid":"22679392","id":"PMC_22679392","title":"The circadian neuropeptide PDF signals preferentially through a specific adenylate cyclase isoform AC3 in M pacemakers of Drosophila.","date":"2012","source":"PLoS biology","url":"https://pubmed.ncbi.nlm.nih.gov/22679392","citation_count":56,"is_preprint":false},{"pmid":"17895882","id":"PMC_17895882","title":"Genetic variation of the adenylyl cyclase 3 (AC3) locus and its influence on type 2 diabetes and obesity susceptibility in Swedish men.","date":"2007","source":"International journal of obesity (2005)","url":"https://pubmed.ncbi.nlm.nih.gov/17895882","citation_count":55,"is_preprint":false},{"pmid":"24113161","id":"PMC_24113161","title":"Upregulation of adenylate cyclase 3 (ADCY3) increases the tumorigenic potential of cells by activating the CREB pathway.","date":"2013","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/24113161","citation_count":44,"is_preprint":false},{"pmid":"36847500","id":"PMC_36847500","title":"Membrane Protein Amuc_1100 Derived from Akkermansia muciniphila Facilitates Lipolysis and Browning via Activating the AC3/PKA/HSL Pathway.","date":"2023","source":"Microbiology spectrum","url":"https://pubmed.ncbi.nlm.nih.gov/36847500","citation_count":42,"is_preprint":false},{"pmid":"2045787","id":"PMC_2045787","title":"Mutagenesis of the AC3 open reading frame of African cassava mosaic virus DNA A reduces DNA B replication and ameliorates disease symptoms.","date":"1991","source":"The Journal of general virology","url":"https://pubmed.ncbi.nlm.nih.gov/2045787","citation_count":34,"is_preprint":false},{"pmid":"21079816","id":"PMC_21079816","title":"Evaluation of the association between the AC3 genetic polymorphisms and obesity in a Chinese Han population.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21079816","citation_count":33,"is_preprint":false},{"pmid":"29921800","id":"PMC_29921800","title":"Interaction between an ADCY3 Genetic Variant and Two Weight-Lowering Diets Affecting Body Fatness and Body Composition Outcomes Depending on Macronutrient Distribution: A Randomized Trial.","date":"2018","source":"Nutrients","url":"https://pubmed.ncbi.nlm.nih.gov/29921800","citation_count":26,"is_preprint":false},{"pmid":"25868729","id":"PMC_25868729","title":"A common sugar-nucleotide-mediated mechanism of inhibition of (glycosamino)glycan biosynthesis, as evidenced by 6F-GalNAc (Ac3).","date":"2015","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/25868729","citation_count":26,"is_preprint":false},{"pmid":"31564937","id":"PMC_31564937","title":"Effects of liraglutide on lipolysis and the AC3/PKA/HSL pathway.","date":"2019","source":"Diabetes, metabolic syndrome and obesity : targets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/31564937","citation_count":23,"is_preprint":false},{"pmid":"21496351","id":"PMC_21496351","title":"The presence of tomato leaf curl Kerala virus AC3 protein enhances viral DNA replication and modulates virus induced gene-silencing mechanism in tomato plants.","date":"2011","source":"Virology journal","url":"https://pubmed.ncbi.nlm.nih.gov/21496351","citation_count":20,"is_preprint":false},{"pmid":"31433213","id":"PMC_31433213","title":"LINC00319 acts as a microRNA-335-5p sponge to accelerate tumor growth and metastasis in gastric cancer by upregulating ADCY3.","date":"2019","source":"American journal of physiology. Gastrointestinal and liver physiology","url":"https://pubmed.ncbi.nlm.nih.gov/31433213","citation_count":20,"is_preprint":false},{"pmid":"20680465","id":"PMC_20680465","title":"AC3-33, a novel secretory protein, inhibits Elk1 transcriptional activity via ERK pathway.","date":"2010","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/20680465","citation_count":18,"is_preprint":false},{"pmid":"34821371","id":"PMC_34821371","title":"Molecular modelling of novel ADCY3 variant predicts a molecular target for tackling obesity.","date":"2021","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34821371","citation_count":17,"is_preprint":false},{"pmid":"20546567","id":"PMC_20546567","title":"Tomato leaf curl Kerala virus (ToLCKeV) AC3 protein forms a higher order oligomer and enhances ATPase activity of replication initiator protein (Rep/AC1).","date":"2010","source":"Virology journal","url":"https://pubmed.ncbi.nlm.nih.gov/20546567","citation_count":16,"is_preprint":false},{"pmid":"38509558","id":"PMC_38509558","title":"Liraglutide improved the reproductive function of obese mice by upregulating the testicular AC3/cAMP/PKA pathway.","date":"2024","source":"Reproductive biology and endocrinology : RB&E","url":"https://pubmed.ncbi.nlm.nih.gov/38509558","citation_count":12,"is_preprint":false},{"pmid":"38334651","id":"PMC_38334651","title":"Postnatal Dynamic Ciliary ARL13B and ADCY3 Localization in the Mouse Brain.","date":"2024","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/38334651","citation_count":11,"is_preprint":false},{"pmid":"38543310","id":"PMC_38543310","title":"Preparation of Nanoparticles Loaded with Membrane-Impermeable Peptide AC3-I and Its Protective Effect on Myocardial Ischemia and Reperfusion.","date":"2024","source":"Pharmaceutics","url":"https://pubmed.ncbi.nlm.nih.gov/38543310","citation_count":8,"is_preprint":false},{"pmid":"38841200","id":"PMC_38841200","title":"ADCY3: the pivotal gene in classical ketogenic diet for the treatment of epilepsy.","date":"2024","source":"Frontiers in cellular neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/38841200","citation_count":6,"is_preprint":false},{"pmid":"34481002","id":"PMC_34481002","title":"ALDH2, ADCY3 and BCMO1 polymorphisms and lifestyle-induced traits are jointly associated with CAD risk in Chinese Han people.","date":"2021","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/34481002","citation_count":5,"is_preprint":false},{"pmid":"36000867","id":"PMC_36000867","title":"Insights into Evolutionary, Genomic, and Biogeographic Characterizations of Chryseobacterium nepalense Represented by a Polyvinyl Alcohol-Degrading Bacterium, AC3.","date":"2022","source":"Microbiology spectrum","url":"https://pubmed.ncbi.nlm.nih.gov/36000867","citation_count":5,"is_preprint":false},{"pmid":"39632398","id":"PMC_39632398","title":"Protein-coding mutation in Adcy3 increases adiposity and alters emotional behaviors sex-dependently in rats.","date":"2024","source":"Obesity (Silver Spring, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/39632398","citation_count":3,"is_preprint":false},{"pmid":"20515784","id":"PMC_20515784","title":"Prokaryotic expression and characterization of human AC3-33 protein.","date":"2010","source":"Frontiers in bioscience (Elite edition)","url":"https://pubmed.ncbi.nlm.nih.gov/20515784","citation_count":3,"is_preprint":false},{"pmid":"39519366","id":"PMC_39519366","title":"Functional Evaluation of a Novel Homozygous ADCY3 Variant Causing Childhood Obesity.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39519366","citation_count":3,"is_preprint":false},{"pmid":"18487146","id":"PMC_18487146","title":"[Molecular cloning and preliminary function study of a novel human gene AC3-33 related to suppress AP-1 activity].","date":"2008","source":"Yi chuan = Hereditas","url":"https://pubmed.ncbi.nlm.nih.gov/18487146","citation_count":3,"is_preprint":false},{"pmid":"31853289","id":"PMC_31853289","title":"Identification and functional analysis of a novel splice variant of AC3-33 in breast cancer.","date":"2019","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31853289","citation_count":3,"is_preprint":false},{"pmid":"37855273","id":"PMC_37855273","title":"A Rare Case of Monogenic Obesity Due to a Novel Variant in the ADCY3 Gene: Challenges in Follow-up and Treatment.","date":"2023","source":"Journal of clinical research in pediatric endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/37855273","citation_count":3,"is_preprint":false},{"pmid":"39707785","id":"PMC_39707785","title":"Network-based meta-analysis and confirmation of genes ATP1A2, FXYD1, and ADCY3 associated with cAMP signaling in breast tumors compared to corresponding normal marginal tissues.","date":"2024","source":"Cellular and molecular biology (Noisy-le-Grand, France)","url":"https://pubmed.ncbi.nlm.nih.gov/39707785","citation_count":2,"is_preprint":false},{"pmid":"38916175","id":"PMC_38916175","title":"Protein-coding mutation in Adcy3 increases adiposity and alters emotional behaviors sex-dependently in rats.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38916175","citation_count":1,"is_preprint":false},{"pmid":"38290616","id":"PMC_38290616","title":"H2S inhibits LiCl/pilocarpine-induced seizures and promotes neuroprotection by regulating TRPV2 expression via the AC3/cAMP/PKA pathway.","date":"2024","source":"Neurochemistry international","url":"https://pubmed.ncbi.nlm.nih.gov/38290616","citation_count":1,"is_preprint":false},{"pmid":"41350952","id":"PMC_41350952","title":"Transcriptome analysis reveals reduced lipid accumulation and mitochondrial metabolic remodeling in ADCY3-overexpressing adipocytes.","date":"2025","source":"Functional & integrative genomics","url":"https://pubmed.ncbi.nlm.nih.gov/41350952","citation_count":1,"is_preprint":false},{"pmid":"37368776","id":"PMC_37368776","title":"A Study of 41 Canine Orthologues of Human Genes Involved in Monogenic Obesity Reveals Marker in the ADCY3 for Body Weight in Labrador Retrievers.","date":"2023","source":"Veterinary sciences","url":"https://pubmed.ncbi.nlm.nih.gov/37368776","citation_count":1,"is_preprint":false},{"pmid":"15172214","id":"PMC_15172214","title":"Induced immunity in Antheraea assama Ww larvae against flacherie causing Pseudomonas aeruginosa AC-3.","date":"2004","source":"Experimental parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/15172214","citation_count":1,"is_preprint":false},{"pmid":"39780573","id":"PMC_39780573","title":"[Association between ADCY3 gene polymorphism and the effects of high-intensity interval training on body composition].","date":"2024","source":"Sheng li xue bao : [Acta physiologica Sinica]","url":"https://pubmed.ncbi.nlm.nih.gov/39780573","citation_count":0,"is_preprint":false},{"pmid":"39060963","id":"PMC_39060963","title":"The association between dietary, physical activity and the DNA methylation of PPARGC1A, HLA-DQA1 and ADCY3 in pregnant women with gestational diabetes mellitus: a nest case-control study.","date":"2024","source":"BMC pregnancy and childbirth","url":"https://pubmed.ncbi.nlm.nih.gov/39060963","citation_count":0,"is_preprint":false},{"pmid":"41494241","id":"PMC_41494241","title":"The rs713586 risk variant dysregulates ADCY3 rather than DNAJC27, leading to obesity through ZFP42-TET1-mediated DNA methylation.","date":"2026","source":"EBioMedicine","url":"https://pubmed.ncbi.nlm.nih.gov/41494241","citation_count":0,"is_preprint":false},{"pmid":"41245346","id":"PMC_41245346","title":"Meta-analysis of GLP1R, GIPR, ADCY3, and CREB1 expression in osteoarthritis identifies CREB1 as a potential biomarker and therapeutic target.","date":"2025","source":"Journal of clinical orthopaedics and trauma","url":"https://pubmed.ncbi.nlm.nih.gov/41245346","citation_count":0,"is_preprint":false},{"pmid":"41630919","id":"PMC_41630919","title":"Circadian ADCY3 Ser107Pro variant bridges difficulty awakening in the morning and adiposity.","date":"2025","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/41630919","citation_count":0,"is_preprint":false},{"pmid":"40766578","id":"PMC_40766578","title":"ADCY3 Ser107Pro links difficulty awakening in the morning to adiposity through circadian regulation of adipose thermogenesis.","date":"2025","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/40766578","citation_count":0,"is_preprint":false},{"pmid":"41542040","id":"PMC_41542040","title":"An Adcy3 coding mutation causes partial loss of enzymatic function, contributing to obesity in a rat model by reducing lipolysis.","date":"2026","source":"Research square","url":"https://pubmed.ncbi.nlm.nih.gov/41542040","citation_count":0,"is_preprint":false},{"pmid":"41398805","id":"PMC_41398805","title":"Assessing the concordance between centromere AC-3 immunofluorescence pattern and anti-centromere protein-B antibody, and analyzing clinical correlates of dual positivity: A retrospective study.","date":"2025","source":"Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/41398805","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.07.28.667339","title":"<i>ADCY3</i>  Ser107Pro links difficulty awakening in the morning to adiposity through circadian regulation of adipose thermogenesis","date":"2025-07-30","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.28.667339","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.11.10.687535","title":"Crosstalk of noradrenergic Ca  <sup>2+</sup>  and cAMP signaling in astrocytes of the murine olfactory bulb","date":"2025-11-11","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.10.687535","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.03.28.645767","title":"A mutation in the transmembrane domain of  <i>Adenylate cyclase 3</i>  impairs enzymatic function to cause sex-specific depression- and anxiety-like behaviors and food seeking in a rat model","date":"2025-04-02","source":"bioRxiv","url":"https://doi.org/10.1101/2025.03.28.645767","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.09.24.614747","title":"ADENYLATE CYCLASE 3 MEDIATES CAROTID BODY ACTIVATION AND AUTONOMIC DYSFUNCTION IN A SLEEP APNEA MODEL","date":"2024-09-26","source":"bioRxiv","url":"https://doi.org/10.1101/2024.09.24.614747","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.10.14.618278","title":"Apoptosis is increased in cortical neurons of female Marfan Syndrome mice","date":"2024-10-17","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.14.618278","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.05.13.25327505","title":"The role of common and rare genetic variation on adiposity across childhood","date":"2025-05-13","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.13.25327505","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":28379,"output_tokens":4539,"usd":0.076611,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12776,"output_tokens":3436,"usd":0.07489,"stage2_stop_reason":"end_turn"},"total_usd":0.151501,"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\": 2018,\n      \"finding\": \"ADCY3 localizes to the primary cilia of neurons, and MC4R colocalizes with ADCY3 at the primary cilia of a subset of hypothalamic neurons. Obesity-associated MC4R mutations impair ciliary localization of MC4R, and inhibition of adenylyl cyclase signaling at the primary cilia of these neurons increases body weight, placing ADCY3-dependent cAMP signaling at neuronal primary cilia as a common pathway underlying genetic causes of obesity.\",\n      \"method\": \"Confocal colocalization imaging in hypothalamic neurons, pharmacological inhibition of adenylyl cyclase in vivo, genetic mouse models with obesity-associated MC4R mutations\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (colocalization, in vivo pharmacology, genetic mutation analysis), replicated context across three concurrent papers in same issue\",\n      \"pmids\": [\"29311635\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Homozygous and compound heterozygous loss-of-function mutations in ADCY3 cause severe monogenic obesity in humans, establishing ADCY3 as a mediator of energy homeostasis. Functional characterization of these mutations confirmed loss of adenylate cyclase 3 activity.\",\n      \"method\": \"Human genetic sequencing, functional characterization of mutations in patient-derived samples\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — functional characterization of mutations combined with human genetics, replicated across independent consanguineous families and different ethnic backgrounds\",\n      \"pmids\": [\"29311637\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"A splice-acceptor-site variant in ADCY3 decreases ADCY3 RNA expression and is associated with markedly increased risk of obesity and type 2 diabetes, confirming ADCY3 loss-of-function as pathogenic for metabolic disease.\",\n      \"method\": \"Population genetics, splice-site variant identification, RNA expression analysis in carriers\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNA expression measured in carriers, single population (Greenlandic), replicated direction in trans-ancestry cohorts\",\n      \"pmids\": [\"29311636\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"ADCY3 overexpression increases cAMP levels and activates the cAMP/PKA/CREB pathway, leading to increased MMP2 and MMP9 mRNA expression and activity. PKA inhibitor treatment decreased MMP2 and MMP9 expression in ADCY3-overexpressing cells, placing ADCY3 upstream of PKA and CREB in this signaling cascade. ADCY3 expression is regulated by promoter CpG methylation.\",\n      \"method\": \"Overexpression and shRNA knockdown in HEK293 and SNU-216 cells, PKA inhibitor treatment, cAMP measurement, luciferase reporter assay, tumor xenograft model, bisulfite sequencing\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (cAMP measurement, PKA inhibition, gene knockdown, in vivo xenograft), single lab\",\n      \"pmids\": [\"24113161\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"In Drosophila M pacemaker neurons, adenylate cyclase AC3 (ortholog of mammalian ADCY3) specifically mediates PDF neuropeptide receptor (PDF-R) signaling to produce cAMP increases. Genetic disruption of AC3 selectively abolishes PDF-induced cAMP responses in M cells without affecting other Gs-coupled GPCR signaling. The AKAP-like scaffolding protein Nervy also reduces PDF responses when knocked down, suggesting PDF-R/AC3 are organized into a 'circadian signalosome'. AC3 is not required for PDF signaling in E pacemaker cells, demonstrating cell-type-specific coupling.\",\n      \"method\": \"Live imaging of intact fly brains with cAMP reporters, transgenic RNAi knockdown, genetic rescue experiments, behavioral analysis\",\n      \"journal\": \"PLoS biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — live in vivo cAMP imaging combined with genetic RNAi and rescue, multiple cell-type comparisons, rigorous controls excluding developmental effects\",\n      \"pmids\": [\"22679392\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ADCY3 localizes dynamically to neuronal primary cilia in the mouse brain in a region- and age-dependent manner. ARL13B+ cilia become relatively rare with age in hypothalamic feeding centers, while ADCY3 becomes a prominent cilia marker in the mature adult brain, indicating dynamic changes in cilia protein composition during postnatal development.\",\n      \"method\": \"Immunofluorescence imaging of mouse brain sections across postnatal ages and brain regions, quantification of cilia marker proportions\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiment with region- and age-dependent quantification, multiple brain regions and time points, single lab\",\n      \"pmids\": [\"38334651\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"A protein-coding mutation in the transmembrane (TM) domain of Adcy3 in rats (Adcy3mut/mut) increases adiposity and alters emotional behaviors in a sex-dependent manner. Male Adcy3mut/mut rats showed increased passive coping and decreased memory, while females showed increased anxiety-like behavior. Adcy3mut/mut males had decreased hypothalamic CREB signaling, with decreased p-AMPK signaling in both sexes, linking the TM domain of ADCY3 to cAMP/CREB and AMPK signaling.\",\n      \"method\": \"CRISPR-SpCas9 TM-domain mutation in WKY rats, high-fat diet feeding, behavioral tests, Western blotting for CREB and p-AMPK\",\n      \"journal\": \"Obesity (Silver Spring, Md.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean CRISPR KO and coding mutation model, multiple behavioral and biochemical readouts, single lab\",\n      \"pmids\": [\"39632398\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The Akkermansia muciniphila membrane protein Amuc_1100 promotes lipolysis and browning in 3T3-L1 preadipocytes by upregulating the AC3/PKA/HSL pathway, increasing lipolysis-related genes (AC3, PKA, HSL) and phosphorylating HSL at Ser660, placing ADCY3 upstream of PKA and HSL in adipocyte lipolysis signaling.\",\n      \"method\": \"Transcriptomics in 3T3-L1 preadipocytes, qPCR, Western blotting, in vivo and in vitro fat accumulation assays\",\n      \"journal\": \"Microbiology spectrum\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — Western blotting and gene expression data only, indirect activation of ADCY3 by a bacterial protein, single lab\",\n      \"pmids\": [\"36847500\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Liraglutide upregulates hepatic AC3 expression and cAMP/PKA activity in obese mice, leading to increased phosphorylated HSL (p-HSL Ser660) and promoting lipolysis via the AC3/PKA/HSL pathway.\",\n      \"method\": \"In vivo liraglutide treatment of obese mice, Western blotting, RT-qPCR, cAMP and PKA activity assays in liver tissue\",\n      \"journal\": \"Diabetes, metabolic syndrome and obesity : targets and therapy\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — in vivo pharmacological upregulation study, indirect activation of ADCY3, single lab, no direct mechanistic perturbation of ADCY3\",\n      \"pmids\": [\"31564937\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In vivo experiments in a rat epilepsy model show that H2S targets and downregulates AC3 expression, thereby modulating the AC3/cAMP/PKA signaling pathway to regulate TRPV2 expression. The AC3 activator forskolin reversed the downregulatory effects of H2S on this pathway, establishing AC3 as a functional node upstream of cAMP/PKA/TRPV2 in seizure regulation.\",\n      \"method\": \"Proteomics in LiCl/Pilocarpine-induced seizure rat model, Western blotting, immunofluorescence, pharmacological rescue with AC3 activator forskolin, EEG and behavioral tests\",\n      \"journal\": \"Neurochemistry international\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — pharmacological rescue with forskolin supports pathway placement, but ADCY3 is identified as a target by proteomics without direct mechanistic perturbation, single lab\",\n      \"pmids\": [\"38290616\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"A novel homozygous nonsense variant in ADCY3 (p.Thr840X) causes severe early-onset obesity. In vitro and in silico functional analyses showed downregulation and impaired activation of the ADCY3 protein, confirming loss of enzymatic activity as the pathogenic mechanism.\",\n      \"method\": \"Gene panel sequencing, in vitro functional assay of ADCY3 activity, in silico protein modeling\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Weak — in vitro enzymatic activity assay combined with in silico modeling, single lab, single patient\",\n      \"pmids\": [\"39519366\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ADCY3 catalyzes the production of cAMP in adipose tissue and is rhythmically expressed there with BMAL1 binding near the Ser107/Pro107 site. A transmembrane domain mutation in Adcy3 causes partial loss of enzymatic function, decreasing cAMP production in response to β-3 adrenergic receptor agonist stimulation in adipose tissue ex vivo, reducing serum free fatty acids and adipose triglyceride lipase expression, and contributing to increased adiposity via decreased lipolytic responsiveness.\",\n      \"method\": \"CRISPR rat model, ex vivo adipose tissue cAMP production assay with β-3 adrenergic receptor agonist, serum FFA measurement, gene expression analysis, cold exposure body temperature measurement\",\n      \"journal\": \"Research square (preprint)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — direct in vitro enzymatic function assay (ex vivo cAMP) combined with genetic loss-of-function rat model and multiple metabolic readouts, single lab, preprint\",\n      \"pmids\": [\"41542040\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In mouse olfactory bulb astrocytes, AC3 (along with AC1) acts downstream of α1 adrenergic receptors as a Ca2+/calmodulin-dependent adenylyl cyclase to produce cAMP in response to norepinephrine/phenylephrine, linking Ca2+ signaling to cAMP production through ADCY3.\",\n      \"method\": \"Live imaging with cAMP and Ca2+ reporters in mouse olfactory bulb astrocytes, pharmacological receptor subtype dissection, Ca2+ depletion experiments\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live imaging with reporters and pharmacological dissection establish pathway order, multiple receptor subtypes tested, single lab, preprint\",\n      \"pmids\": [\"bio_10.1101_2025.11.10.687535\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In a mouse carotid body model of chronic intermittent hypoxia (CIH), Adcy3-dependent cAMP production is downstream of Olfr78 activation by H2S. CIH increases cAMP in the carotid body, absent in Adcy3, Cth, and Olfr78 null mice. CIH-induced cAMP via ADCY3 mediates enhanced Ca2+ influx through cyclic nucleotide-gated channels (CNGA2). Adcy3 null mice do not exhibit carotid body activation or autonomic dysfunction in response to CIH.\",\n      \"method\": \"Adcy3 null mouse model, cAMP measurement in carotid body, Ca2+ imaging in glomus cells, comparison across Adcy3, Olfr78, Cth, and Cnga2 mutant mice\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple genetic null models with biochemical and cellular readouts establish pathway position, single lab, preprint\",\n      \"pmids\": [\"bio_10.1101_2024.09.24.614747\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"The obesity risk SNP rs713586-C allele reduces ADCY3 promoter activity via impaired ZFP42 transcription factor binding, leading to decreased TET1 recruitment and increased DNA methylation at the ADCY3 enhancer and promoter regions, suppressing ADCY3 expression. Dnajc27 knockout mice did not develop obesity, excluding DNAJC27 as the functional target of rs713586, while ADCY3 downregulation was confirmed as the relevant mechanism.\",\n      \"method\": \"Dual-luciferase reporter assay, CRISPR/Cas9 genome editing in cell lines, Dnajc27 knockout mice, bisulfite sequencing for DNA methylation, ZFP42-TET1 interaction analysis\",\n      \"journal\": \"EBioMedicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (reporter assay, genome editing, KO mice, DNA methylation, TF binding), single lab but comprehensive mechanistic dissection\",\n      \"pmids\": [\"41494241\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"A transmembrane domain mutation in Adcy3 decreases hypothalamic cAMP production in rats without altering ADCY3 membrane content, demonstrating that the TM domain is required for enzymatic (cAMP-generating) function. Adcy3mut/mut rats showed sex-specific depression- and anxiety-like behaviors and food seeking, with increased leptin levels in males.\",\n      \"method\": \"CRISPR rat model, hypothalamic cAMP measurement, membrane fractionation and Western blotting, comprehensive behavioral testing\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct cAMP measurement in hypothalamus combined with membrane fractionation and behavioral battery, single lab, preprint\",\n      \"pmids\": [\"bio_10.1101_2025.03.28.645767\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ADCY3 overexpression in adipocytes impairs adipogenesis by downregulating the adipogenic transcription factors CEBPα and PPARγ, reduces lipid droplet number and size by suppressing triglyceride synthesis and fatty acid metabolism genes (DGAT1, DGAT2, ACC, SCD, FASN, ACSL1), and suppresses oxidative phosphorylation genes through PPARγ signaling.\",\n      \"method\": \"ADCY3 overexpression in adipocytes, transcriptomic profiling, gene expression and protein analysis (qPCR, Western blotting), lipid droplet quantification\",\n      \"journal\": \"Functional & integrative genomics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — overexpression study with transcriptomics and gene expression, single lab, no direct measurement of cAMP or ADCY3 enzymatic activity\",\n      \"pmids\": [\"41350952\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ADCY3 is a transmembrane adenylyl cyclase that localizes to neuronal primary cilia (especially in hypothalamic feeding centers) and catalyzes conversion of ATP to cAMP; it acts downstream of Gs-coupled GPCRs (including MC4R and olfactory receptor Olfr78) and Ca2+/calmodulin signaling to generate cAMP, which activates PKA/CREB and PKA/HSL signaling to regulate energy homeostasis, lipolysis, and behavior, with loss-of-function causing severe obesity in humans and rodents through impaired cAMP-mediated signaling in hypothalamic cilia, adipose tissue, and other metabolically relevant tissues.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ADCY3 is a transmembrane adenylyl cyclase that catalyzes ATP-to-cAMP conversion at neuronal primary cilia and in metabolic tissues, functioning as a central node linking Gs-coupled GPCR and Ca2+ signaling to cAMP-dependent control of energy homeostasis and behavior [#0, #1]. In the hypothalamus, ADCY3 localizes to the primary cilia of feeding-center neurons where it colocalizes with MC4R, and disruption of ciliary adenylyl cyclase signaling increases body weight, defining ADCY3-dependent ciliary cAMP as a convergence point for genetic obesity [#0]; its enzymatic function requires the transmembrane domain, as a TM-domain mutation reduces hypothalamic cAMP production without altering membrane content [#15]. Loss-of-function in humans causes severe monogenic obesity: homozygous and compound heterozygous mutations, a nonsense variant (p.Thr840X), and regulatory variants that reduce ADCY3 expression all abolish or diminish adenylyl cyclase activity and confer obesity and type 2 diabetes risk [#1, #2, #10]. The obesity risk allele rs713586-C suppresses ADCY3 transcription through impaired ZFP42 binding, reduced TET1 recruitment, and increased promoter/enhancer DNA methylation, with DNAJC27 excluded as the functional target [#14]. Downstream, ADCY3-generated cAMP activates PKA to drive CREB-dependent transcription and HSL (Ser660)-dependent lipolysis in adipose and hepatic tissue, where ADCY3 is rhythmically expressed and required for beta-3 adrenergic lipolytic responsiveness [#3, #11]. ADCY3 also operates as a Ca2+/calmodulin-dependent cyclase downstream of alpha1 adrenergic receptors in astrocytes [#12] and, in Drosophila pacemaker neurons, mediates cell-type-specific PDF receptor signaling within a putative circadian signalosome [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Established that ADCY3 is not a generic cAMP source but couples selectively to specific GPCRs in a cell-type-restricted manner, introducing the concept of a localized signalosome.\",\n      \"evidence\": \"Live cAMP imaging with transgenic RNAi and rescue in Drosophila M and E pacemaker neurons\",\n      \"pmids\": [\"22679392\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Done in fly ortholog; mammalian cell-type selectivity not directly demonstrated here\", \"Molecular basis of receptor-cyclase coupling specificity unresolved\", \"Role of Nervy/AKAP scaffold in mammals untested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Placed ADCY3 upstream of the cAMP/PKA/CREB cascade as a driver of downstream transcriptional output, and showed its expression is epigenetically controlled.\",\n      \"evidence\": \"Overexpression/knockdown, PKA inhibition, cAMP and luciferase assays, xenograft, and bisulfite sequencing in HEK293 and gastric cancer cells\",\n      \"pmids\": [\"24113161\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Cancer cell context, not metabolic tissue\", \"MMP induction is downstream and indirect\", \"Methylation regulation not linked to physiological signal\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Resolved the cellular site and physiological role of ADCY3 in obesity by localizing it to hypothalamic neuronal primary cilia alongside MC4R, defining ciliary cAMP as a common obesity pathway.\",\n      \"evidence\": \"Confocal colocalization, in vivo adenylyl cyclase inhibition, and genetic mouse models of MC4R mutations\",\n      \"pmids\": [\"29311635\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct demonstration that MC4R signals through ADCY3 enzymatically not shown\", \"Identity of full ciliary receptor repertoire incomplete\", \"Downstream neuronal effectors of ciliary cAMP not defined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established ADCY3 loss-of-function as causal for human monogenic obesity, converting a candidate into a validated disease gene.\",\n      \"evidence\": \"Human sequencing across consanguineous families plus functional characterization confirming loss of cyclase activity; independent population genetics on a splice variant reducing RNA expression\",\n      \"pmids\": [\"29311637\", \"29311636\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Splice-variant finding limited to single population (Medium confidence)\", \"Tissue-specific contribution of each loss-of-function mechanism not dissected\", \"Genotype-phenotype dose relationship incomplete\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Localized the enzymatic function to the transmembrane domain and showed ADCY3 ciliary occupancy changes dynamically with brain region and age, refining how and where cAMP is generated.\",\n      \"evidence\": \"CRISPR TM-domain mutant rats with hypothalamic cAMP measurement and membrane fractionation; immunofluorescence of cilia markers across mouse brain ages\",\n      \"pmids\": [\"39632398\", \"bio_10.1101_2025.03.28.645767\", \"38334651\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of TM-domain requirement for catalysis unresolved\", \"Functional consequence of age-dependent ciliary remodeling untested\", \"Sex-specific behavioral mechanisms unexplained\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Positioned ADCY3 downstream of olfactory receptor Olfr78 and of Ca2+/calmodulin signaling, extending its role beyond MC4R to chemosensory and Ca2+-coupled cAMP generation.\",\n      \"evidence\": \"Adcy3 null and Olfr78/Cth/Cnga2 mutant mice in carotid body CIH model; live cAMP/Ca2+ imaging with receptor dissection in olfactory bulb astrocytes\",\n      \"pmids\": [\"bio_10.1101_2024.09.24.614747\", \"bio_10.1101_2025.11.10.687535\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Both are preprints\", \"Direct physical coupling of Olfr78/alpha1-AR to ADCY3 not shown\", \"Generalizability beyond carotid body and astrocytes unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined ADCY3's peripheral metabolic role by showing it drives beta-3 adrenergic, rhythmically regulated cAMP/PKA/HSL lipolysis in adipose and liver, with overexpression conversely impairing adipogenesis.\",\n      \"evidence\": \"Ex vivo adipose cAMP assay in CRISPR rats with BMAL1-binding analysis; liraglutide and Amuc_1100 pathway studies; ADCY3-overexpression adipocyte transcriptomics\",\n      \"pmids\": [\"41542040\", \"31564937\", \"36847500\", \"41350952\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Adipose mechanism partly preprint and partly Low-confidence indirect activation studies\", \"Direct cAMP/enzymatic measurement absent in several adipocyte studies\", \"Reconciliation of overexpression vs loss-of-function phenotypes incomplete\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Mechanistically explained a major obesity GWAS signal by tracing the rs713586-C allele to ZFP42/TET1-dependent DNA methylation that suppresses ADCY3 expression, and excluded the neighboring DNAJC27.\",\n      \"evidence\": \"Dual-luciferase assays, CRISPR editing, Dnajc27 knockout mice, bisulfite sequencing, and ZFP42-TET1 interaction analysis\",\n      \"pmids\": [\"41494241\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo demonstration of allele-specific methylation in human tissue limited\", \"Tissue specificity of the regulatory effect not fully mapped\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How ADCY3 achieves receptor- and tissue-specific coupling, and the structural basis by which the transmembrane domain governs catalysis, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure-function model linking TM domain to active site\", \"Mechanism of selective GPCR-ADCY3 pairing across cell types unknown\", \"Direct physical interactome of ADCY3 not defined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0009975\", \"supporting_discovery_ids\": [1, 3, 10, 11, 15]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [3, 11, 15]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 5]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [15]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 4, 12, 13]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [1, 11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"MC4R\", \"Olfr78\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}