{"gene":"GUCY1B1","run_date":"2026-04-28T18:06:53","timeline":{"discoveries":[{"year":1990,"finding":"GUCY1B1 (GC-S beta 2) encodes a 76.3-kDa soluble guanylyl cyclase subunit that is preferentially expressed in kidney and liver, distinct from the lung/brain-expressed GC-S beta 1, and its C-terminus contains a consensus isoprenylation/carboxymethylation sequence (-C-V-V-L), establishing heterogeneity among heterodimeric soluble guanylyl cyclase forms.","method":"PCR cloning of cDNA using conserved catalytic domain sequences, sequence analysis, and mRNA tissue distribution","journal":"Biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 — original cloning with sequence-based functional inference; single lab, foundational identification","pmids":["1980215"],"is_preprint":false},{"year":2011,"finding":"Notch signaling directly induces transcription of both GUCY1A3 and GUCY1B3 (sGC heterodimer subunits), and NO-sGC signaling activated through this Notch-dependent autocrine loop is necessary to drive endothelial-to-mesenchymal transition (EndMT) in the developing atrioventricular canal.","method":"Genetic/transcriptional analysis in embryonic heart development model; Notch gain-of-function, NO/cGMP pathway inhibition, and EndMT phenotypic readout","journal":"Developmental cell","confidence":"Medium","confidence_rationale":"Tier 2 — pathway placement by epistasis and loss-of-function; single lab with multiple orthogonal approaches","pmids":["21839921"],"is_preprint":false},{"year":2013,"finding":"Cell-specific deletion of the β1 subunit of NO-sensitive guanylyl cyclase (GUCY1B3) from smooth muscle cells (SMCs) incompletely reduced NO-induced fundus relaxation, while deletion from interstitial cells of Cajal (ICCs) alone had little effect; combined deletion from both SMCs and ICCs abolished nitrergic relaxation and increased gut transit time, establishing that ICCs and SMCs jointly mediate nitrergic GI smooth muscle relaxation through sGC.","method":"Cell-specific conditional knockout mice (Cre-lox), isometric force studies, in vivo gut transit measurement","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 1-2 — clean cell-specific KO with defined cellular and in vivo phenotypic readout, multiple genetic conditions tested","pmids":["23528627"],"is_preprint":false},{"year":2013,"finding":"Notch3 activation in immortalized ovarian surface epithelial cells increases GUCY1B3 expression, NO-induced cGMP production, and PKG expression, thereby enhancing NO/cGMP-induced phosphorylation of VASP (a direct PKG substrate); conversely, Notch inhibition with DAPT reduces GUCY1B3 expression and downstream signaling in OVCAR3 ovarian cancer cells.","method":"Forced Notch3 expression, gamma-secretase inhibitor (DAPT) treatment, cGMP production assay, VASP phosphorylation western blot, sGC inhibitor (ODQ) treatment with proliferation assay","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods in single lab establishing Notch→GUCY1B3→PKG→VASP pathway","pmids":["24041655"],"is_preprint":false},{"year":2017,"finding":"Hypertension (angiotensin II treatment) represses GUCY1B3 expression in mouse aorta via the Notch signaling pathway; Notch coactivators FRYL and MAML2 are required for constitutive sGC expression, and JAG/NOTCH gain- and loss-of-function experiments demonstrated that Notch signaling directly controls sGC (GUCY1A3/GUCY1B3) expression in vascular smooth muscle.","method":"AngII hypertension mouse model, gene expression analysis, transcription factor binding motif analysis, Notch gain/loss-of-function experiments, western blotting, immunohistochemistry, RNA-Seq in human coronary artery","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — gain and loss of function with mechanistic follow-up across mouse, rat, and human data; single lab","pmids":["28465505"],"is_preprint":false},{"year":2018,"finding":"GUCY1B3 (sGCβ subunit) exerts cardioprotective effects against ischemia-reperfusion injury via PKCε/Akt signaling: GUCY1B3 overexpression restored IR-induced cell death in neonatal rat ventricular myocytes, and GUCY1B3 silencing in a mouse coronary ligation model aggravated cardiac dysfunction and increased infarct size with inactivation of PKCε and Akt.","method":"Overexpression and siRNA knockdown in cardiomyocytes, specific PKC/Akt inhibitors, in vivo mouse myocardial infarction model with echocardiography and histology","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 — KO/OE with defined phenotype and pathway placement via pharmacological inhibitors; single lab","pmids":["30485489"],"is_preprint":false},{"year":2022,"finding":"FoxO4 is a critical transcriptional activator of sGCβ (GUCY1B3) in vascular smooth muscle cells: FoxO4 knockdown decreased Gucy1b3 mRNA by ~50% and sGCβ protein by ~50%, reduced cGMP production and PKG-dependent phosphorylation >50%, and chromatin immunoprecipitation showed FoxO4 directly binds FoxO DNA motifs in the GUCY1B3 promoter in human aortic SMCs.","method":"shRNA knockdown in rat aortic SMCs, promoter luciferase assay, chromatin immunoprecipitation (ChIP), cGMP production assay, western blotting","journal":"American journal of physiology. Heart and circulatory physiology","confidence":"High","confidence_rationale":"Tier 1-2 — direct binding demonstrated by ChIP, functional promoter assay, and phenotypic rescue; multiple orthogonal methods in single lab","pmids":["35089807"],"is_preprint":false},{"year":2023,"finding":"Co-overexpression of exogenous GUCY1A3 and GUCY1B3 subunits in HEK293 cells forms functional soluble guanylyl cyclase enzyme with increased sGC activity and elevated cGMP levels, demonstrating that the two subunits can assemble into a catalytically active heterodimer when co-expressed.","method":"Lentiviral overexpression of GUCY1A3 and GUCY1B3 in HEK293 cells, sGC enzyme activity assay, cGMP measurement","journal":"Technology and health care","confidence":"Medium","confidence_rationale":"Tier 2 — direct enzymatic reconstitution in mammalian cells with functional cGMP readout; single lab","pmids":["36442224"],"is_preprint":false},{"year":2025,"finding":"GUCY1B1 (sGC β subunit) is expressed in retinal vascular cells and neuronal elements (retinal ganglion, bipolar, and amacrine cells); oxidative stress impairs sGC function in retinal cells, and the sGC activator runcaciguat restores sGC activity and improves neuroretinal function and morphology in rat ischemia-reperfusion and streptozotocin-induced diabetic models.","method":"Immunohistochemistry for subunit localization, in vitro oxidative stress assays, electroretinography, optokinetic tracking, retinal morphometry in rat models","journal":"Diabetes","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization with functional consequence demonstrated in multiple in vivo models; single lab","pmids":["40249725"],"is_preprint":false},{"year":2026,"finding":"In acute lung injury, ASIC1a negatively regulates LncRNA00178, which acts as a ceRNA to sponge miR-466b-3p; miR-466b-3p directly targets and degrades Gucy1b1 mRNA (validated by dual-luciferase reporter assay), and ASIC1a-mediated suppression of this pathway reduces Gucy1b1 expression and promotes alveolar epithelial cell apoptosis.","method":"High-throughput RNA sequencing, dual-luciferase reporter gene assay (confirming miR-466b-3p binding to Gucy1b1 3'UTR), plasmid transfection for gain/loss of function in LPS-stimulated RLE-6TN cells, apoptosis assays","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2 — direct target validation by luciferase assay with epistasis via ceRNA pathway; single lab","pmids":["41830717"],"is_preprint":false}],"current_model":"GUCY1B1/GUCY1B3 encodes the β1 subunit of soluble guanylyl cyclase (sGC), which heterodimerizes with the α subunit (GUCY1A1/GUCY1A3) to form the principal nitric oxide receptor that catalyzes cGMP synthesis; its transcription is positively regulated by FoxO4 and Notch/MAML2 signaling in vascular smooth muscle, and its downstream cGMP/PKG signaling mediates vascular relaxation (jointly through smooth muscle cells and interstitial cells of Cajal in the GI tract), cardiac protection via PKCε/Akt, and endothelial-to-mesenchymal transition during cardiac valve development, while being suppressed in hypertension and acute lung injury through distinct regulatory mechanisms."},"narrative":{"teleology":[{"year":1990,"claim":"Cloning of GUCY1B1 established that multiple soluble guanylyl cyclase β subunits exist with distinct tissue expression, resolving prior uncertainty about sGC heterogeneity.","evidence":"PCR cloning from human kidney/liver cDNA with sequence analysis and tissue mRNA distribution","pmids":["1980215"],"confidence":"Medium","gaps":["No functional enzymatic characterization of the β2 isoform was performed","Whether this subunit forms active heterodimers with distinct α subunits was not tested","The isoprenylation motif identified at the C-terminus was not functionally validated"]},{"year":2011,"claim":"Demonstration that Notch signaling directly induces GUCY1B3 transcription and that the resulting NO-sGC-cGMP axis is required for endothelial-to-mesenchymal transition placed sGC as a developmental effector downstream of Notch in cardiac valve formation.","evidence":"Notch gain-of-function and NO/cGMP pathway inhibition in embryonic atrioventricular canal development model","pmids":["21839921"],"confidence":"Medium","gaps":["Direct Notch binding to the GUCY1B3 promoter was not shown by ChIP","Whether cGMP acts through PKG or other effectors in EndMT was not resolved"]},{"year":2013,"claim":"Cell-specific knockouts revealed that nitrergic GI relaxation requires sGC in both smooth muscle cells and interstitial cells of Cajal acting in concert, resolving the long-standing question of which cell type mediates NO-dependent gut motility.","evidence":"Conditional Cre-lox deletion of Gucy1b3 from SMCs and/or ICCs with isometric force and gut transit measurements in mice","pmids":["23528627"],"confidence":"High","gaps":["The relative cGMP output from each cell type was not quantified","Whether ICC-specific sGC contributes to relaxation in other GI regions was not tested"]},{"year":2013,"claim":"Notch3-driven upregulation of GUCY1B3 and downstream PKG/VASP phosphorylation in ovarian epithelial cells extended the Notch–sGC axis beyond cardiac development to an epithelial/cancer context.","evidence":"Notch3 overexpression and DAPT inhibition in ovarian surface epithelial and OVCAR3 cancer cells with cGMP and pVASP readouts","pmids":["24041655"],"confidence":"Medium","gaps":["Direct Notch3 binding to the GUCY1B3 promoter was not demonstrated","Functional consequences for cancer cell proliferation were shown with sGC inhibition but not with specific GUCY1B3 knockdown"]},{"year":2017,"claim":"Identification of MAML2 and FRYL as Notch coactivators required for constitutive sGC expression in vascular smooth muscle, and demonstration that angiotensin II-induced hypertension represses sGC through the Notch pathway, established a disease-relevant transcriptional circuit controlling GUCY1B3.","evidence":"AngII hypertension mouse model, Notch gain/loss-of-function, motif analysis, western blot, and RNA-Seq in human coronary artery","pmids":["28465505"],"confidence":"Medium","gaps":["Direct ChIP evidence for MAML2/Notch binding at the GUCY1B3 locus was not provided","Whether Notch-dependent sGC downregulation is reversible upon antihypertensive treatment is unknown"]},{"year":2018,"claim":"Overexpression and silencing experiments established that GUCY1B3 protects cardiomyocytes from ischemia-reperfusion injury through PKCε/Akt signaling, identifying a cGMP-independent or cGMP-to-kinase cardioprotective branch downstream of sGC.","evidence":"sGC overexpression/siRNA in neonatal rat ventricular myocytes and in vivo mouse coronary ligation with pharmacological PKC/Akt inhibition","pmids":["30485489"],"confidence":"Medium","gaps":["Whether the PKCε/Akt pathway is activated by cGMP/PKG or through a non-canonical mechanism was not determined","Single-lab finding without independent replication"]},{"year":2022,"claim":"ChIP and promoter assays demonstrated that FoxO4 directly binds FoxO motifs in the GUCY1B3 promoter and is required for ~50% of basal sGC β expression and cGMP/PKG signaling in vascular smooth muscle, identifying the first transcription factor with validated direct binding at this locus.","evidence":"ChIP, luciferase promoter assay, shRNA knockdown in rat and human aortic SMCs with cGMP and PKG readouts","pmids":["35089807"],"confidence":"High","gaps":["Interplay between FoxO4 and Notch at the GUCY1B3 promoter was not tested","Whether FoxO4 regulation is altered in hypertension is unknown"]},{"year":2023,"claim":"Reconstitution of catalytically active sGC by co-expression of GUCY1A3 and GUCY1B3 in HEK293 cells provided direct biochemical proof that these two subunits are sufficient for heterodimer formation and cGMP production.","evidence":"Lentiviral co-overexpression in HEK293 cells with sGC activity and cGMP measurement","pmids":["36442224"],"confidence":"Medium","gaps":["NO-stimulated versus basal activity was not characterized","Stoichiometry and heme incorporation were not assessed"]},{"year":2025,"claim":"Localization of GUCY1B1 in retinal vascular and neuronal cells and demonstration that oxidative stress impairs sGC function—rescuable by pharmacological sGC activation—extended the functional role of the β1 subunit to neuroretinal protection in diabetic and ischemic retinopathy.","evidence":"Immunohistochemistry, oxidative stress assays, electroretinography, and optokinetic tracking in rat diabetic and ischemia-reperfusion models","pmids":["40249725"],"confidence":"Medium","gaps":["Cell-type-specific genetic deletion in the retina was not performed","Whether oxidative stress acts by heme oxidation of sGC was not directly tested"]},{"year":2026,"claim":"Identification of a ceRNA regulatory axis (LncRNA00178–miR-466b-3p–Gucy1b1) downstream of ASIC1a in acute lung injury revealed a post-transcriptional mechanism that degrades Gucy1b1 mRNA and promotes alveolar epithelial apoptosis.","evidence":"RNA-seq, dual-luciferase reporter assay validating miR-466b-3p targeting of Gucy1b1 3′UTR, gain/loss-of-function in LPS-stimulated rat alveolar epithelial cells","pmids":["41830717"],"confidence":"Medium","gaps":["Single-lab finding in rat cells without in vivo validation of the ceRNA axis","Whether this regulatory mechanism operates in human lung injury is unknown","Downstream consequences of Gucy1b1 loss in this context were limited to apoptosis readouts"]},{"year":null,"claim":"Key unresolved questions include the structural basis for β1-specific functions versus other β isoforms, the integration of FoxO4 and Notch transcriptional inputs at the GUCY1B3 promoter, and whether sGC β1-specific knockout (distinct from pan-sGC disruption) produces unique phenotypes in the cardiovascular or nervous systems.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural model of the full-length β1 subunit in complex with the α subunit exists from the primary literature surveyed","Isoform-specific functions of β1 versus β2 remain largely undefined","Cross-talk between transcriptional (Notch, FoxO4) and post-transcriptional (miRNA) regulation of GUCY1B1 has not been explored"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0009975","term_label":"cyclase activity","supporting_discovery_ids":[0,2,7]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,7]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,2,3,5,6]},{"term_id":"R-HSA-397014","term_label":"Muscle contraction","supporting_discovery_ids":[2]}],"complexes":["soluble guanylyl cyclase (sGC) heterodimer"],"partners":["GUCY1A1","FOXO4","MAML2","NOTCH3"],"other_free_text":[]},"mechanistic_narrative":"GUCY1B1 encodes the β1 subunit of soluble guanylyl cyclase (sGC), which heterodimerizes with the α1 subunit (GUCY1A1) to form the principal intracellular receptor for nitric oxide (NO) and catalyze the synthesis of cGMP from GTP [PMID:1980215, PMID:36442224]. In the vasculature and gastrointestinal tract, sGC-derived cGMP signals through PKG to mediate smooth muscle relaxation, with cell-specific knockout studies demonstrating that both smooth muscle cells and interstitial cells of Cajal are jointly required for nitrergic GI relaxation [PMID:23528627]. Transcription of GUCY1B1 in vascular smooth muscle is directly activated by FoxO4 binding at the promoter and by Notch/MAML2 signaling, and hypertension suppresses GUCY1B1 expression through impaired Notch activity [PMID:35089807, PMID:28465505]. During cardiac development, Notch-driven induction of sGC subunits enables NO/cGMP-dependent endothelial-to-mesenchymal transition, while in cardiomyocytes GUCY1B1 confers protection against ischemia-reperfusion injury via PKCε/Akt signaling [PMID:21839921, PMID:30485489]."},"prefetch_data":{"uniprot":{"accession":"Q02153","full_name":"Guanylate cyclase soluble subunit beta-1","aliases":["Guanylate cyclase soluble subunit beta-3","GCS-beta-3","Soluble guanylate cyclase small subunit"],"length_aa":619,"mass_kda":70.5,"function":"Mediates responses to nitric oxide (NO) by catalyzing the biosynthesis of the signaling molecule cGMP","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q02153/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/GUCY1B1","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/GUCY1B1","total_profiled":1310},"omim":[{"mim_id":"605140","title":"CHAPERONIN CONTAINING T-COMPLEX POLYPEPTIDE 1, SUBUNIT 7; CCT7","url":"https://www.omim.org/entry/605140"},{"mim_id":"139397","title":"GUANYLATE CYCLASE, SOLUBLE, BETA-1; GUCY1B1","url":"https://www.omim.org/entry/139397"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/GUCY1B1"},"hgnc":{"alias_symbol":["GC-SB3","GC-S-beta-1"],"prev_symbol":["GUC1B3","GUCY1B3"]},"alphafold":{"accession":"Q02153","domains":[{"cath_id":"3.90.1520.10","chopping":"2-115_142-158","consensus_level":"medium","plddt":88.3395,"start":2,"end":158},{"cath_id":"3.30.450.260","chopping":"210-286_303-336","consensus_level":"high","plddt":89.6294,"start":210,"end":336},{"cath_id":"3.30.70.1230","chopping":"411-605","consensus_level":"high","plddt":91.318,"start":411,"end":605}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q02153","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q02153-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q02153-F1-predicted_aligned_error_v6.png","plddt_mean":85.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=GUCY1B1","jax_strain_url":"https://www.jax.org/strain/search?query=GUCY1B1"},"sequence":{"accession":"Q02153","fasta_url":"https://rest.uniprot.org/uniprotkb/Q02153.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q02153/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q02153"}},"corpus_meta":[{"pmid":"1980215","id":"PMC_1980215","title":"A new form of guanylyl cyclase is preferentially expressed in rat kidney.","date":"1990","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/1980215","citation_count":164,"is_preprint":false},{"pmid":"21839921","id":"PMC_21839921","title":"Notch initiates the endothelial-to-mesenchymal transition in the atrioventricular canal through autocrine activation of soluble guanylyl cyclase.","date":"2011","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/21839921","citation_count":138,"is_preprint":false},{"pmid":"17050858","id":"PMC_17050858","title":"Placental growth throughout the last two thirds of pregnancy in sheep: vascular development and angiogenic factor expression.","date":"2006","source":"Biology of reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/17050858","citation_count":120,"is_preprint":false},{"pmid":"18432406","id":"PMC_18432406","title":"ICF, an immunodeficiency syndrome: DNA methyltransferase 3B involvement, chromosome anomalies, and gene dysregulation.","date":"2008","source":"Autoimmunity","url":"https://pubmed.ncbi.nlm.nih.gov/18432406","citation_count":103,"is_preprint":false},{"pmid":"20663985","id":"PMC_20663985","title":"Sildenafil treatment in vivo stimulates Leydig cell steroidogenesis via the cAMP/cGMP signaling pathway.","date":"2010","source":"American journal of physiology. Endocrinology and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/20663985","citation_count":64,"is_preprint":false},{"pmid":"22506031","id":"PMC_22506031","title":"Complex control of GABA(A) receptor subunit mRNA expression: variation, covariation, and genetic regulation.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22506031","citation_count":50,"is_preprint":false},{"pmid":"23528627","id":"PMC_23528627","title":"Cell-specific deletion of nitric oxide-sensitive guanylyl cyclase reveals a dual pathway for nitrergic neuromuscular transmission in the murine fundus.","date":"2013","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/23528627","citation_count":48,"is_preprint":false},{"pmid":"17526946","id":"PMC_17526946","title":"Effects of estradiol-17beta on expression of mRNA for seven angiogenic factors and their receptors in the endometrium of ovariectomized (OVX) ewes.","date":"2006","source":"Endocrine","url":"https://pubmed.ncbi.nlm.nih.gov/17526946","citation_count":46,"is_preprint":false},{"pmid":"28465505","id":"PMC_28465505","title":"Hypertension reduces soluble guanylyl cyclase expression in the mouse aorta via the Notch signaling pathway.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28465505","citation_count":37,"is_preprint":false},{"pmid":"17008469","id":"PMC_17008469","title":"Expression of endothelial nitric oxide synthase in the ovine ovary throughout the estrous cycle.","date":"2006","source":"Reproduction (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/17008469","citation_count":33,"is_preprint":false},{"pmid":"15201957","id":"PMC_15201957","title":"Inhibition of angiogenesis in human glioma cell lines by antisense RNA from the soluble guanylate cyclase genes, GUCY1A3 and GUCY1B3.","date":"2004","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/15201957","citation_count":32,"is_preprint":false},{"pmid":"16943586","id":"PMC_16943586","title":"Isolation and characterization of ovine luteal pericytes and effects of nitric oxide on pericyte expression of angiogenic factors.","date":"2006","source":"Endocrine","url":"https://pubmed.ncbi.nlm.nih.gov/16943586","citation_count":25,"is_preprint":false},{"pmid":"24041655","id":"PMC_24041655","title":"Notch activation augments nitric oxide/soluble guanylyl cyclase signaling in immortalized ovarian surface epithelial cells and ovarian cancer cells.","date":"2013","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/24041655","citation_count":20,"is_preprint":false},{"pmid":"37291580","id":"PMC_37291580","title":"Exploring the molecular mechanism of comorbidity of autism spectrum disorder and inflammatory bowel disease by combining multiple data sets.","date":"2023","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/37291580","citation_count":18,"is_preprint":false},{"pmid":"28532842","id":"PMC_28532842","title":"Effects of diet and arginine treatment during the luteal phase on ovarian NO/PGC-1α signaling in ewes.","date":"2017","source":"Theriogenology","url":"https://pubmed.ncbi.nlm.nih.gov/28532842","citation_count":16,"is_preprint":false},{"pmid":"27915366","id":"PMC_27915366","title":"Aging has the opposite effect on cAMP and cGMP circadian variations in rat Leydig cells.","date":"2016","source":"Journal of comparative physiology. 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cancer","date":"2024-08-08","source":"bioRxiv","url":"https://doi.org/10.1101/2024.08.06.606802","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":27018,"output_tokens":2791,"usd":0.06146},"stage2":{"model":"claude-opus-4-6","input_tokens":6150,"output_tokens":2939,"usd":0.156337},"total_usd":0.217797,"stage1_batch_id":"msgbatch_01DzrsgVgYngY1hvM3DU1yF2","stage2_batch_id":"msgbatch_01KA9b7iRzKFdhoBUT7vCvSe","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1990,\n      \"finding\": \"GUCY1B1 (GC-S beta 2) encodes a 76.3-kDa soluble guanylyl cyclase subunit that is preferentially expressed in kidney and liver, distinct from the lung/brain-expressed GC-S beta 1, and its C-terminus contains a consensus isoprenylation/carboxymethylation sequence (-C-V-V-L), establishing heterogeneity among heterodimeric soluble guanylyl cyclase forms.\",\n      \"method\": \"PCR cloning of cDNA using conserved catalytic domain sequences, sequence analysis, and mRNA tissue distribution\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — original cloning with sequence-based functional inference; single lab, foundational identification\",\n      \"pmids\": [\"1980215\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Notch signaling directly induces transcription of both GUCY1A3 and GUCY1B3 (sGC heterodimer subunits), and NO-sGC signaling activated through this Notch-dependent autocrine loop is necessary to drive endothelial-to-mesenchymal transition (EndMT) in the developing atrioventricular canal.\",\n      \"method\": \"Genetic/transcriptional analysis in embryonic heart development model; Notch gain-of-function, NO/cGMP pathway inhibition, and EndMT phenotypic readout\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pathway placement by epistasis and loss-of-function; single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"21839921\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Cell-specific deletion of the β1 subunit of NO-sensitive guanylyl cyclase (GUCY1B3) from smooth muscle cells (SMCs) incompletely reduced NO-induced fundus relaxation, while deletion from interstitial cells of Cajal (ICCs) alone had little effect; combined deletion from both SMCs and ICCs abolished nitrergic relaxation and increased gut transit time, establishing that ICCs and SMCs jointly mediate nitrergic GI smooth muscle relaxation through sGC.\",\n      \"method\": \"Cell-specific conditional knockout mice (Cre-lox), isometric force studies, in vivo gut transit measurement\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — clean cell-specific KO with defined cellular and in vivo phenotypic readout, multiple genetic conditions tested\",\n      \"pmids\": [\"23528627\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Notch3 activation in immortalized ovarian surface epithelial cells increases GUCY1B3 expression, NO-induced cGMP production, and PKG expression, thereby enhancing NO/cGMP-induced phosphorylation of VASP (a direct PKG substrate); conversely, Notch inhibition with DAPT reduces GUCY1B3 expression and downstream signaling in OVCAR3 ovarian cancer cells.\",\n      \"method\": \"Forced Notch3 expression, gamma-secretase inhibitor (DAPT) treatment, cGMP production assay, VASP phosphorylation western blot, sGC inhibitor (ODQ) treatment with proliferation assay\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods in single lab establishing Notch→GUCY1B3→PKG→VASP pathway\",\n      \"pmids\": [\"24041655\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Hypertension (angiotensin II treatment) represses GUCY1B3 expression in mouse aorta via the Notch signaling pathway; Notch coactivators FRYL and MAML2 are required for constitutive sGC expression, and JAG/NOTCH gain- and loss-of-function experiments demonstrated that Notch signaling directly controls sGC (GUCY1A3/GUCY1B3) expression in vascular smooth muscle.\",\n      \"method\": \"AngII hypertension mouse model, gene expression analysis, transcription factor binding motif analysis, Notch gain/loss-of-function experiments, western blotting, immunohistochemistry, RNA-Seq in human coronary artery\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — gain and loss of function with mechanistic follow-up across mouse, rat, and human data; single lab\",\n      \"pmids\": [\"28465505\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"GUCY1B3 (sGCβ subunit) exerts cardioprotective effects against ischemia-reperfusion injury via PKCε/Akt signaling: GUCY1B3 overexpression restored IR-induced cell death in neonatal rat ventricular myocytes, and GUCY1B3 silencing in a mouse coronary ligation model aggravated cardiac dysfunction and increased infarct size with inactivation of PKCε and Akt.\",\n      \"method\": \"Overexpression and siRNA knockdown in cardiomyocytes, specific PKC/Akt inhibitors, in vivo mouse myocardial infarction model with echocardiography and histology\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KO/OE with defined phenotype and pathway placement via pharmacological inhibitors; single lab\",\n      \"pmids\": [\"30485489\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"FoxO4 is a critical transcriptional activator of sGCβ (GUCY1B3) in vascular smooth muscle cells: FoxO4 knockdown decreased Gucy1b3 mRNA by ~50% and sGCβ protein by ~50%, reduced cGMP production and PKG-dependent phosphorylation >50%, and chromatin immunoprecipitation showed FoxO4 directly binds FoxO DNA motifs in the GUCY1B3 promoter in human aortic SMCs.\",\n      \"method\": \"shRNA knockdown in rat aortic SMCs, promoter luciferase assay, chromatin immunoprecipitation (ChIP), cGMP production assay, western blotting\",\n      \"journal\": \"American journal of physiology. Heart and circulatory physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct binding demonstrated by ChIP, functional promoter assay, and phenotypic rescue; multiple orthogonal methods in single lab\",\n      \"pmids\": [\"35089807\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Co-overexpression of exogenous GUCY1A3 and GUCY1B3 subunits in HEK293 cells forms functional soluble guanylyl cyclase enzyme with increased sGC activity and elevated cGMP levels, demonstrating that the two subunits can assemble into a catalytically active heterodimer when co-expressed.\",\n      \"method\": \"Lentiviral overexpression of GUCY1A3 and GUCY1B3 in HEK293 cells, sGC enzyme activity assay, cGMP measurement\",\n      \"journal\": \"Technology and health care\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct enzymatic reconstitution in mammalian cells with functional cGMP readout; single lab\",\n      \"pmids\": [\"36442224\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"GUCY1B1 (sGC β subunit) is expressed in retinal vascular cells and neuronal elements (retinal ganglion, bipolar, and amacrine cells); oxidative stress impairs sGC function in retinal cells, and the sGC activator runcaciguat restores sGC activity and improves neuroretinal function and morphology in rat ischemia-reperfusion and streptozotocin-induced diabetic models.\",\n      \"method\": \"Immunohistochemistry for subunit localization, in vitro oxidative stress assays, electroretinography, optokinetic tracking, retinal morphometry in rat models\",\n      \"journal\": \"Diabetes\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization with functional consequence demonstrated in multiple in vivo models; single lab\",\n      \"pmids\": [\"40249725\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"In acute lung injury, ASIC1a negatively regulates LncRNA00178, which acts as a ceRNA to sponge miR-466b-3p; miR-466b-3p directly targets and degrades Gucy1b1 mRNA (validated by dual-luciferase reporter assay), and ASIC1a-mediated suppression of this pathway reduces Gucy1b1 expression and promotes alveolar epithelial cell apoptosis.\",\n      \"method\": \"High-throughput RNA sequencing, dual-luciferase reporter gene assay (confirming miR-466b-3p binding to Gucy1b1 3'UTR), plasmid transfection for gain/loss of function in LPS-stimulated RLE-6TN cells, apoptosis assays\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct target validation by luciferase assay with epistasis via ceRNA pathway; single lab\",\n      \"pmids\": [\"41830717\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"GUCY1B1/GUCY1B3 encodes the β1 subunit of soluble guanylyl cyclase (sGC), which heterodimerizes with the α subunit (GUCY1A1/GUCY1A3) to form the principal nitric oxide receptor that catalyzes cGMP synthesis; its transcription is positively regulated by FoxO4 and Notch/MAML2 signaling in vascular smooth muscle, and its downstream cGMP/PKG signaling mediates vascular relaxation (jointly through smooth muscle cells and interstitial cells of Cajal in the GI tract), cardiac protection via PKCε/Akt, and endothelial-to-mesenchymal transition during cardiac valve development, while being suppressed in hypertension and acute lung injury through distinct regulatory mechanisms.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"GUCY1B1 encodes the β1 subunit of soluble guanylyl cyclase (sGC), which heterodimerizes with the α1 subunit (GUCY1A1) to form the principal intracellular receptor for nitric oxide (NO) and catalyze the synthesis of cGMP from GTP [PMID:1980215, PMID:36442224]. In the vasculature and gastrointestinal tract, sGC-derived cGMP signals through PKG to mediate smooth muscle relaxation, with cell-specific knockout studies demonstrating that both smooth muscle cells and interstitial cells of Cajal are jointly required for nitrergic GI relaxation [PMID:23528627]. Transcription of GUCY1B1 in vascular smooth muscle is directly activated by FoxO4 binding at the promoter and by Notch/MAML2 signaling, and hypertension suppresses GUCY1B1 expression through impaired Notch activity [PMID:35089807, PMID:28465505]. During cardiac development, Notch-driven induction of sGC subunits enables NO/cGMP-dependent endothelial-to-mesenchymal transition, while in cardiomyocytes GUCY1B1 confers protection against ischemia-reperfusion injury via PKCε/Akt signaling [PMID:21839921, PMID:30485489].\",\n  \"teleology\": [\n    {\n      \"year\": 1990,\n      \"claim\": \"Cloning of GUCY1B1 established that multiple soluble guanylyl cyclase β subunits exist with distinct tissue expression, resolving prior uncertainty about sGC heterogeneity.\",\n      \"evidence\": \"PCR cloning from human kidney/liver cDNA with sequence analysis and tissue mRNA distribution\",\n      \"pmids\": [\"1980215\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No functional enzymatic characterization of the β2 isoform was performed\",\n        \"Whether this subunit forms active heterodimers with distinct α subunits was not tested\",\n        \"The isoprenylation motif identified at the C-terminus was not functionally validated\"\n      ]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Demonstration that Notch signaling directly induces GUCY1B3 transcription and that the resulting NO-sGC-cGMP axis is required for endothelial-to-mesenchymal transition placed sGC as a developmental effector downstream of Notch in cardiac valve formation.\",\n      \"evidence\": \"Notch gain-of-function and NO/cGMP pathway inhibition in embryonic atrioventricular canal development model\",\n      \"pmids\": [\"21839921\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct Notch binding to the GUCY1B3 promoter was not shown by ChIP\",\n        \"Whether cGMP acts through PKG or other effectors in EndMT was not resolved\"\n      ]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Cell-specific knockouts revealed that nitrergic GI relaxation requires sGC in both smooth muscle cells and interstitial cells of Cajal acting in concert, resolving the long-standing question of which cell type mediates NO-dependent gut motility.\",\n      \"evidence\": \"Conditional Cre-lox deletion of Gucy1b3 from SMCs and/or ICCs with isometric force and gut transit measurements in mice\",\n      \"pmids\": [\"23528627\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"The relative cGMP output from each cell type was not quantified\",\n        \"Whether ICC-specific sGC contributes to relaxation in other GI regions was not tested\"\n      ]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Notch3-driven upregulation of GUCY1B3 and downstream PKG/VASP phosphorylation in ovarian epithelial cells extended the Notch–sGC axis beyond cardiac development to an epithelial/cancer context.\",\n      \"evidence\": \"Notch3 overexpression and DAPT inhibition in ovarian surface epithelial and OVCAR3 cancer cells with cGMP and pVASP readouts\",\n      \"pmids\": [\"24041655\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct Notch3 binding to the GUCY1B3 promoter was not demonstrated\",\n        \"Functional consequences for cancer cell proliferation were shown with sGC inhibition but not with specific GUCY1B3 knockdown\"\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identification of MAML2 and FRYL as Notch coactivators required for constitutive sGC expression in vascular smooth muscle, and demonstration that angiotensin II-induced hypertension represses sGC through the Notch pathway, established a disease-relevant transcriptional circuit controlling GUCY1B3.\",\n      \"evidence\": \"AngII hypertension mouse model, Notch gain/loss-of-function, motif analysis, western blot, and RNA-Seq in human coronary artery\",\n      \"pmids\": [\"28465505\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct ChIP evidence for MAML2/Notch binding at the GUCY1B3 locus was not provided\",\n        \"Whether Notch-dependent sGC downregulation is reversible upon antihypertensive treatment is unknown\"\n      ]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Overexpression and silencing experiments established that GUCY1B3 protects cardiomyocytes from ischemia-reperfusion injury through PKCε/Akt signaling, identifying a cGMP-independent or cGMP-to-kinase cardioprotective branch downstream of sGC.\",\n      \"evidence\": \"sGC overexpression/siRNA in neonatal rat ventricular myocytes and in vivo mouse coronary ligation with pharmacological PKC/Akt inhibition\",\n      \"pmids\": [\"30485489\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether the PKCε/Akt pathway is activated by cGMP/PKG or through a non-canonical mechanism was not determined\",\n        \"Single-lab finding without independent replication\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"ChIP and promoter assays demonstrated that FoxO4 directly binds FoxO motifs in the GUCY1B3 promoter and is required for ~50% of basal sGC β expression and cGMP/PKG signaling in vascular smooth muscle, identifying the first transcription factor with validated direct binding at this locus.\",\n      \"evidence\": \"ChIP, luciferase promoter assay, shRNA knockdown in rat and human aortic SMCs with cGMP and PKG readouts\",\n      \"pmids\": [\"35089807\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Interplay between FoxO4 and Notch at the GUCY1B3 promoter was not tested\",\n        \"Whether FoxO4 regulation is altered in hypertension is unknown\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Reconstitution of catalytically active sGC by co-expression of GUCY1A3 and GUCY1B3 in HEK293 cells provided direct biochemical proof that these two subunits are sufficient for heterodimer formation and cGMP production.\",\n      \"evidence\": \"Lentiviral co-overexpression in HEK293 cells with sGC activity and cGMP measurement\",\n      \"pmids\": [\"36442224\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"NO-stimulated versus basal activity was not characterized\",\n        \"Stoichiometry and heme incorporation were not assessed\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Localization of GUCY1B1 in retinal vascular and neuronal cells and demonstration that oxidative stress impairs sGC function—rescuable by pharmacological sGC activation—extended the functional role of the β1 subunit to neuroretinal protection in diabetic and ischemic retinopathy.\",\n      \"evidence\": \"Immunohistochemistry, oxidative stress assays, electroretinography, and optokinetic tracking in rat diabetic and ischemia-reperfusion models\",\n      \"pmids\": [\"40249725\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Cell-type-specific genetic deletion in the retina was not performed\",\n        \"Whether oxidative stress acts by heme oxidation of sGC was not directly tested\"\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identification of a ceRNA regulatory axis (LncRNA00178–miR-466b-3p–Gucy1b1) downstream of ASIC1a in acute lung injury revealed a post-transcriptional mechanism that degrades Gucy1b1 mRNA and promotes alveolar epithelial apoptosis.\",\n      \"evidence\": \"RNA-seq, dual-luciferase reporter assay validating miR-466b-3p targeting of Gucy1b1 3′UTR, gain/loss-of-function in LPS-stimulated rat alveolar epithelial cells\",\n      \"pmids\": [\"41830717\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Single-lab finding in rat cells without in vivo validation of the ceRNA axis\",\n        \"Whether this regulatory mechanism operates in human lung injury is unknown\",\n        \"Downstream consequences of Gucy1b1 loss in this context were limited to apoptosis readouts\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis for β1-specific functions versus other β isoforms, the integration of FoxO4 and Notch transcriptional inputs at the GUCY1B3 promoter, and whether sGC β1-specific knockout (distinct from pan-sGC disruption) produces unique phenotypes in the cardiovascular or nervous systems.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural model of the full-length β1 subunit in complex with the α subunit exists from the primary literature surveyed\",\n        \"Isoform-specific functions of β1 versus β2 remain largely undefined\",\n        \"Cross-talk between transcriptional (Notch, FoxO4) and post-transcriptional (miRNA) regulation of GUCY1B1 has not been explored\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0009975\", \"supporting_discovery_ids\": [0, 2, 7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 2, 3, 5, 6]},\n      {\"term_id\": \"R-HSA-397014\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"complexes\": [\n      \"soluble guanylyl cyclase (sGC) heterodimer\"\n    ],\n    \"partners\": [\n      \"GUCY1A1\",\n      \"FOXO4\",\n      \"MAML2\",\n      \"NOTCH3\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}