{"gene":"GUCY1B1","run_date":"2026-06-10T01:55:21","timeline":{"discoveries":[{"year":1990,"finding":"GUCY1B1 (GC-S beta 1) encodes a 70-kDa subunit of the soluble guanylyl cyclase heterodimer, preferentially expressed in lung and brain; a related beta subunit (GC-S beta 2) is preferentially expressed in kidney and liver, demonstrating heterogeneity among heterodimeric forms of guanylyl cyclase.","method":"PCR-based cDNA cloning, amino acid sequence analysis, mRNA expression profiling across tissues","journal":"Biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct molecular cloning and tissue expression characterization in a single focused study","pmids":["1980215"],"is_preprint":false},{"year":2011,"finding":"Notch activation induces transcription of both sGC subunits GUCY1A3 and GUCY1B3 (the nitric oxide receptor heterodimer) during endothelial-to-mesenchymal transition in the atrioventricular canal; this Notch-dependent autocrine NO/sGC loop is necessary to drive early EndMT in valve development.","method":"Genetic and pharmacological activation/inhibition of Notch signaling in vivo and in vitro, gene expression analysis, functional assays of EndMT","journal":"Developmental cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal approaches (gain/loss of function, gene expression, functional readout) in a single lab study","pmids":["21839921"],"is_preprint":false},{"year":2013,"finding":"GUCY1B3 (β1 subunit of NO-sensitive guanylyl cyclase) is expressed in both smooth muscle cells (SMCs) and interstitial cells of Cajal (ICCs) of the murine fundus; deletion from either SMCs or ICCs alone incompletely reduced NO-induced relaxation, but combined SMC/ICC-specific deletion completely abolished nitrergic relaxation and increased gut transit time, demonstrating that ICCs and SMCs jointly mediate enteric NO signaling.","method":"Cell-specific conditional knockout (Cre-lox) of GUCY1B3 in SMCs, ICCs, or both; isometric force studies; gut transit time measurement","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 2 / Strong — cell-specific KO with multiple orthogonal phenotypic readouts (isometric force, gut transit), rigorous genetic controls","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 phosphorylation of VASP (a direct PKG substrate); conversely, Notch inhibition by DAPT reduces GUCY1B3 expression and NO-induced cGMP production, placing Notch as a positive upstream regulator of NO/sGC signaling.","method":"Forced Notch3 activation and DAPT-mediated Notch inhibition in IOSE and ovarian cancer cells; cGMP production assay; VASP phosphorylation western blot; sGC inhibitor ODQ cell growth assay","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (gain/loss of function, cGMP assay, phosphorylation assay) in a single lab","pmids":["24041655"],"is_preprint":false},{"year":2017,"finding":"Hypertension-induced (angiotensin II) repression of GUCY1B3/GUCY1A3 in the mouse aorta occurs via the Notch signaling pathway; the Notch coactivators FRYL and MAML2 are reduced by hypertension, and gain/loss-of-function experiments demonstrated that JAG/NOTCH signaling together with MAML2 and FRYL controls sGC expression. Reduced sGC was associated with curtailed NO-dependent vasorelaxation.","method":"Angiotensin II hypertension model in mice; gene expression analysis (Western blot, immunohistochemistry); transcription factor binding motif analysis; gain and loss of Notch signaling; functional vasorelaxation assays; RNA-Seq in human coronary artery","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (gain/loss of function, protein expression, functional vasorelaxation), replicated across mouse, rat, and human data","pmids":["28465505"],"is_preprint":false},{"year":2022,"finding":"FoxO4 is a critical transcriptional activator of GUCY1B3 (sGCβ) in vascular smooth muscle cells (SMCs); FoxO4 knockdown caused ~50% decrease in GUCY1B3 mRNA and sGCβ protein, with >50% loss of cGMP production and PKG-dependent phosphorylation. FoxO4 directly binds FoxO DNA motifs in the GUCY1B3 promoter. FoxO1 or FoxO3 knockdown paradoxically increased GUCY1B3 expression.","method":"shRNA knockdown of FoxO family members in rat aortic SMCs; qRT-PCR; Western blot; cGMP production assay; promoter luciferase assay; chromatin immunoprecipitation (ChIP) in human aortic SMCs","journal":"American journal of physiology. Heart and circulatory physiology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — direct ChIP showing FoxO4 binding to GUCY1B3 promoter, combined with promoter luciferase, KD functional assays (cGMP, phosphorylation), multiple orthogonal methods in one rigorous study","pmids":["35089807"],"is_preprint":false},{"year":2018,"finding":"GUCY1B3 (β subunit of sGC) exerts cardioprotective effects against ischemia-reperfusion (IR) injury; GUCY1B3 was upregulated in neonatal rat ventricular myocytes after IR, overexpression reduced IR-induced cell death and apoptosis, and these effects were mediated via PKCε/Akt signaling. In a mouse coronary ligation model, GUCY1B3 silencing aggravated cardiac dysfunction and infarct size in association with inactivation of PKCε and Akt.","method":"GUCY1B3 overexpression and siRNA silencing in neonatal rat ventricular myocytes; in vitro IR model; specific PKCδ, PKCε, and Akt inhibitors; mouse coronary artery ligation model with GUCY1B3 knockdown; echocardiography, apoptosis assays","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — both in vitro and in vivo gain/loss-of-function with pharmacological pathway dissection, single lab study","pmids":["30485489"],"is_preprint":false},{"year":2004,"finding":"GUCY1B3 (β subunit of sGC) participates in mediating cGMP production upstream of VEGF expression in glioma cells; antisense knockdown of GUCY1B3 in human glioma cell lines (CCF-STTG1 and U-87MG) markedly reduced cGMP content, VEGF expression, and in vitro angiogenic activity (HUVEC growth induction), and dramatically suppressed subcutaneous tumor formation and vascular index in nude mice.","method":"Antisense RNA transfection in glioma cell lines; cGMP measurement; VEGF expression analysis; HUVEC proliferation assay; xenograft tumor formation assay with vascular index measurement","journal":"Oncology reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with multiple in vitro and in vivo readouts in a single lab study","pmids":["15201957"],"is_preprint":false},{"year":2023,"finding":"Co-expression of GUCY1A3 and GUCY1B3 subunits via lentiviral vectors in HEK293 cells resulted in formation of functional sGC enzyme with increased catalytic activity and elevated cellular cGMP levels, demonstrating that co-expression of the two subunits is sufficient for assembly of a functional heterodimeric enzyme.","method":"Lentiviral overexpression of GUCY1A3 and GUCY1B3 in HEK293 cells; sGC enzyme activity assay; cGMP quantification; cytotoxicity assay","journal":"Technology and health care","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro reconstitution of enzyme activity via co-expression, single lab, single study","pmids":["36442224"],"is_preprint":false},{"year":2025,"finding":"GUCY1B1 is expressed in retinal vascular cells and neuronal elements (retinal ganglion, bipolar, and amacrine cells) in healthy human and rodent retinas; sGC function (requiring GUCY1B1) is impaired by oxidative stress in the retina, and the sGC activator runcaciguat activates sGC in multiple retinal cell types, counteracting oxidative stress-induced damage and improving neuroretinal function and morphology in rat retinal ischemia-reperfusion and streptozotocin-diabetic models.","method":"Immunohistochemistry for sGC subunit localization; in vitro and in vivo sGC activity assays; electroretinography; optokinetic tracking; retinal morphology assessment in rat IR and STZ-diabetic models","journal":"Diabetes","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiments combined with functional in vivo and in vitro assays, multiple orthogonal methods in one study","pmids":["40249725"],"is_preprint":false},{"year":2021,"finding":"In zebrafish, combined morpholino downregulation of both gucy1a1 and gucy1b1 (but not individual knockdown of either alone) increased blood flow and linear velocity, indicating that both subunits together are required for sGC-mediated regulation of vascular blood flow. An sGC stimulator (BAY41-2272) rescued impaired cGMP production in gucy1a1 mutant larvae.","method":"CRISPR-Cas9 gucy1a1 mutant zebrafish; individual and combined morpholino knockdown of gucy1a1 and gucy1b1; blood flow parameter measurement; pharmacological sGC stimulation; cGMP assay","journal":"Frontiers in physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and morpholino loss-of-function with functional blood flow readouts, multiple genetic conditions tested in single lab study","pmids":["33716783"],"is_preprint":false},{"year":2025,"finding":"In LPS-stimulated alveolar epithelial cells (rat RLE-6TN), ASIC1a negatively regulates LncRNA00178, which in turn acts as a sponge for miR-466b-3p; miR-466b-3p targets Gucy1b1 mRNA for degradation. ASIC1a thus suppresses Gucy1b1 expression via the LncRNA00178/miR-466b-3p axis, promoting apoptosis of alveolar epithelial cells in acute lung injury.","method":"High-throughput transcriptomic sequencing of rat lung tissue; plasmid transfection to alter ASIC1a, LncRNA00178, miR-466b-3p, Gucy1b1 expression in RLE-6TN cells; dual-luciferase reporter assay; apoptosis assays","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dual-luciferase validation of miRNA-target interaction and direct manipulation of pathway components in a single lab study; note that this involves non-coding RNA regulation of the canonical protein","pmids":["41830717"],"is_preprint":false}],"current_model":"GUCY1B1 (alias GUCY1B3) encodes the β1 subunit of soluble guanylyl cyclase (sGC), an obligate heterodimer with the α1 subunit (GUCY1A1/GUCY1A3) that functions as the main intracellular receptor for nitric oxide (NO); upon NO binding, the heterodimer catalyzes cGMP synthesis to mediate vasodilation, cardiac protection (via PKCε/Akt), gastrointestinal smooth muscle relaxation (through SMCs and ICCs jointly), and retinal neuroprotection—with its transcription positively regulated by FoxO4 and Notch/MAML2/FRYL signaling and negatively regulated by hypertension-induced Notch pathway suppression and miR-466b-3p targeting."},"narrative":{"mechanistic_narrative":"GUCY1B1 (β1 subunit, alias GUCY1B3) encodes one of the two obligate subunits of soluble guanylyl cyclase (sGC), the nitric oxide (NO) receptor that synthesizes cGMP; co-expression of GUCY1B1 with the α1 subunit (GUCY1A3) in heterologous cells is sufficient to assemble a catalytically active, cGMP-producing heterodimer [PMID:1980215, PMID:36442224]. Functional output requires both subunits in concert: in vivo, only combined loss of α1 and β1 abolishes sGC-dependent vascular and smooth-muscle responses, whereas knockdown of either alone is insufficient [PMID:33716783]. Through this NO/sGC/cGMP axis, GUCY1B1 mediates a range of tissue-level processes—nitrergic gastrointestinal smooth muscle relaxation, in which it operates jointly in smooth muscle cells and interstitial cells of Cajal so that both populations are required for complete relaxation and normal gut transit [PMID:23528627]; cardioprotection against ischemia-reperfusion injury via downstream PKCε/Akt signaling [PMID:30485489]; and retinal neuroprotection against oxidative stress in vascular and neuronal cells [PMID:40249725]. Downstream signaling proceeds through cGMP-dependent protein kinase (PKG) and its substrate VASP [PMID:24041655, PMID:35089807]. GUCY1B1 transcription is positively controlled by FoxO4, which binds FoxO motifs in the GUCY1B3 promoter, and by Notch signaling acting through coactivators MAML2 and FRYL, while hypertension represses sGC by suppressing this Notch pathway [PMID:28465505, PMID:35089807]; post-transcriptionally, Gucy1b1 mRNA is targeted for degradation by miR-466b-3p [PMID:41830717]. In tumor and developmental settings the same axis is co-opted, contributing to glioma angiogenesis via cGMP-dependent VEGF expression [PMID:15201957] and to Notch-driven endothelial-to-mesenchymal transition during cardiac valve development [PMID:21839921].","teleology":[{"year":1990,"claim":"Established that GUCY1B1 encodes a distinct ~70-kDa β subunit of the soluble guanylyl cyclase heterodimer with tissue-selective expression, defining the molecular identity of the gene.","evidence":"PCR-based cDNA cloning and tissue mRNA expression profiling","pmids":["1980215"],"confidence":"Medium","gaps":["Did not demonstrate enzymatic activity of the reconstituted heterodimer","Subunit stoichiometry and NO-binding role not yet assigned"]},{"year":2004,"claim":"Linked GUCY1B1-dependent cGMP production to a pro-angiogenic program, showing the axis can drive VEGF expression and tumor vascularization in glioma.","evidence":"Antisense knockdown in human glioma cell lines with cGMP, VEGF, HUVEC angiogenesis, and xenograft readouts","pmids":["15201957"],"confidence":"Medium","gaps":["Mechanism linking cGMP to VEGF transcription not resolved","Single-gene antisense without rescue"]},{"year":2011,"claim":"Placed GUCY1B1 within a Notch-driven autocrine NO/sGC loop required for endothelial-to-mesenchymal transition in valve development, extending its role beyond mature smooth muscle.","evidence":"Genetic/pharmacological Notch manipulation in vivo and in vitro with EndMT functional assays","pmids":["21839921"],"confidence":"Medium","gaps":["Direct transcriptional control of GUCY1B1 promoter by Notch not demonstrated here","Relative contribution of β1 vs α1 not separated"]},{"year":2013,"claim":"Defined the cellular basis of nitrergic gastrointestinal relaxation, showing GUCY1B1 acts jointly in smooth muscle cells and interstitial cells of Cajal so that both are required for complete NO-induced relaxation.","evidence":"Cell-specific conditional knockout of GUCY1B3 in SMCs, ICCs, or both, with isometric force and gut transit measurements","pmids":["23528627"],"confidence":"High","gaps":["cGMP effector steps downstream in each cell type not dissected","Does not address other smooth muscle beds"]},{"year":2013,"claim":"Connected Notch signaling to GUCY1B1 transcription and downstream PKG/VASP output, positioning Notch as a positive upstream regulator of the NO/sGC pathway.","evidence":"Notch3 activation and DAPT inhibition in ovarian epithelial/cancer cells with cGMP, PKG, and VASP phosphorylation readouts","pmids":["24041655"],"confidence":"Medium","gaps":["No direct promoter occupancy by Notch effectors shown","Whether effect is direct or indirect unresolved"]},{"year":2017,"claim":"Identified the Notch coactivators MAML2 and FRYL as controllers of sGC expression and showed hypertension represses GUCY1B1 by suppressing this Notch axis, curtailing NO-dependent vasorelaxation.","evidence":"Angiotensin II hypertension model with gain/loss of Notch signaling, protein expression, and vasorelaxation assays across mouse, rat, and human data","pmids":["28465505"],"confidence":"Medium","gaps":["Direct binding of MAML2/FRYL complex to the GUCY1B1 promoter not shown","Causality of repression in human hypertension inferred from expression"]},{"year":2018,"claim":"Established a cardioprotective function for GUCY1B1 against ischemia-reperfusion injury and traced it to PKCε/Akt signaling.","evidence":"Overexpression and siRNA silencing in neonatal rat ventricular myocytes plus mouse coronary ligation, with pharmacological PKC/Akt inhibitors and echocardiography","pmids":["30485489"],"confidence":"Medium","gaps":["How sGC/cGMP couples to PKCε/Akt activation mechanistically not defined","Single lab; α1 subunit involvement not addressed"]},{"year":2021,"claim":"Demonstrated genetically that both subunits together are required for sGC-mediated vascular blood flow regulation, confirming obligate heterodimer function in vivo.","evidence":"CRISPR mutant and combined/individual morpholino knockdown of gucy1a1 and gucy1b1 in zebrafish with blood flow measurement and pharmacological sGC stimulation rescue","pmids":["33716783"],"confidence":"Medium","gaps":["Morpholino off-target effects inherent to method","cGMP measurement performed in α1 mutant, not β1-specific"]},{"year":2022,"claim":"Identified FoxO4 as a direct transcriptional activator of GUCY1B1 in vascular smooth muscle, with opposing roles for FoxO1/FoxO3, establishing isoform-specific transcriptional control of the gene.","evidence":"shRNA knockdown of FoxO family members in aortic SMCs with qRT-PCR, cGMP, promoter luciferase, and ChIP","pmids":["35089807"],"confidence":"High","gaps":["Mechanism of opposing FoxO1/FoxO3 effect not resolved","Interplay with Notch regulation not integrated"]},{"year":2023,"claim":"Confirmed by reconstitution that co-expression of α1 and β1 subunits alone is sufficient to form a catalytically active cGMP-producing enzyme.","evidence":"Lentiviral co-expression of GUCY1A3 and GUCY1B3 in HEK293 cells with sGC activity and cGMP assays","pmids":["36442224"],"confidence":"Medium","gaps":["No structural or stoichiometric characterization","NO-responsiveness of the reconstituted enzyme not tested here"]},{"year":2025,"claim":"Extended GUCY1B1 function to retinal neuroprotection, localizing sGC to retinal vascular and neuronal cells and showing pharmacological sGC activation counteracts oxidative-stress damage.","evidence":"Immunohistochemistry plus in vitro/in vivo sGC activity assays and functional retinal readouts in rat IR and STZ-diabetic models","pmids":["40249725"],"confidence":"Medium","gaps":["β1-specific genetic loss-of-function in retina not performed","Downstream cGMP effectors in neurons not identified"]},{"year":2025,"claim":"Revealed post-transcriptional control of Gucy1b1 by a non-coding RNA network in alveolar epithelial injury, with miR-466b-3p directly targeting its mRNA.","evidence":"Transcriptomic sequencing, pathway component manipulation, and dual-luciferase reporter assay in LPS-stimulated rat alveolar epithelial cells","pmids":["41830717"],"confidence":"Medium","gaps":["sGC enzymatic/cGMP consequence of Gucy1b1 repression not measured","Single lab; ASIC1a/lncRNA axis not validated in vivo"]},{"year":null,"claim":"How the multiple converging transcriptional (FoxO4, Notch/MAML2/FRYL) and post-transcriptional (miR-466b-3p) regulators are integrated to set GUCY1B1 levels across tissues, and the structural basis of NO sensing by the β1 subunit, remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model linking the distinct regulatory inputs","No structural characterization of the β1 NO-binding module in the timeline","Tissue-specific effector pathways downstream of cGMP only partially mapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0009975","term_label":"cyclase activity","supporting_discovery_ids":[0,8]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[8]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[8]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,8,10]}],"complexes":["soluble guanylyl cyclase (sGC) heterodimer"],"partners":["GUCY1A3"],"other_free_text":[]}},"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":140,"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":105,"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. 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cDNA cloning, amino acid sequence analysis, mRNA expression profiling across tissues\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct molecular cloning and tissue expression characterization in a single focused study\",\n      \"pmids\": [\"1980215\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Notch activation induces transcription of both sGC subunits GUCY1A3 and GUCY1B3 (the nitric oxide receptor heterodimer) during endothelial-to-mesenchymal transition in the atrioventricular canal; this Notch-dependent autocrine NO/sGC loop is necessary to drive early EndMT in valve development.\",\n      \"method\": \"Genetic and pharmacological activation/inhibition of Notch signaling in vivo and in vitro, gene expression analysis, functional assays of EndMT\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal approaches (gain/loss of function, gene expression, functional readout) in a single lab study\",\n      \"pmids\": [\"21839921\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"GUCY1B3 (β1 subunit of NO-sensitive guanylyl cyclase) is expressed in both smooth muscle cells (SMCs) and interstitial cells of Cajal (ICCs) of the murine fundus; deletion from either SMCs or ICCs alone incompletely reduced NO-induced relaxation, but combined SMC/ICC-specific deletion completely abolished nitrergic relaxation and increased gut transit time, demonstrating that ICCs and SMCs jointly mediate enteric NO signaling.\",\n      \"method\": \"Cell-specific conditional knockout (Cre-lox) of GUCY1B3 in SMCs, ICCs, or both; isometric force studies; gut transit time measurement\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cell-specific KO with multiple orthogonal phenotypic readouts (isometric force, gut transit), rigorous genetic controls\",\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 phosphorylation of VASP (a direct PKG substrate); conversely, Notch inhibition by DAPT reduces GUCY1B3 expression and NO-induced cGMP production, placing Notch as a positive upstream regulator of NO/sGC signaling.\",\n      \"method\": \"Forced Notch3 activation and DAPT-mediated Notch inhibition in IOSE and ovarian cancer cells; cGMP production assay; VASP phosphorylation western blot; sGC inhibitor ODQ cell growth assay\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (gain/loss of function, cGMP assay, phosphorylation assay) in a single lab\",\n      \"pmids\": [\"24041655\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Hypertension-induced (angiotensin II) repression of GUCY1B3/GUCY1A3 in the mouse aorta occurs via the Notch signaling pathway; the Notch coactivators FRYL and MAML2 are reduced by hypertension, and gain/loss-of-function experiments demonstrated that JAG/NOTCH signaling together with MAML2 and FRYL controls sGC expression. Reduced sGC was associated with curtailed NO-dependent vasorelaxation.\",\n      \"method\": \"Angiotensin II hypertension model in mice; gene expression analysis (Western blot, immunohistochemistry); transcription factor binding motif analysis; gain and loss of Notch signaling; functional vasorelaxation assays; RNA-Seq in human coronary artery\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (gain/loss of function, protein expression, functional vasorelaxation), replicated across mouse, rat, and human data\",\n      \"pmids\": [\"28465505\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"FoxO4 is a critical transcriptional activator of GUCY1B3 (sGCβ) in vascular smooth muscle cells (SMCs); FoxO4 knockdown caused ~50% decrease in GUCY1B3 mRNA and sGCβ protein, with >50% loss of cGMP production and PKG-dependent phosphorylation. FoxO4 directly binds FoxO DNA motifs in the GUCY1B3 promoter. FoxO1 or FoxO3 knockdown paradoxically increased GUCY1B3 expression.\",\n      \"method\": \"shRNA knockdown of FoxO family members in rat aortic SMCs; qRT-PCR; Western blot; cGMP production assay; promoter luciferase assay; chromatin immunoprecipitation (ChIP) in human aortic SMCs\",\n      \"journal\": \"American journal of physiology. Heart and circulatory physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — direct ChIP showing FoxO4 binding to GUCY1B3 promoter, combined with promoter luciferase, KD functional assays (cGMP, phosphorylation), multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"35089807\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"GUCY1B3 (β subunit of sGC) exerts cardioprotective effects against ischemia-reperfusion (IR) injury; GUCY1B3 was upregulated in neonatal rat ventricular myocytes after IR, overexpression reduced IR-induced cell death and apoptosis, and these effects were mediated via PKCε/Akt signaling. In a mouse coronary ligation model, GUCY1B3 silencing aggravated cardiac dysfunction and infarct size in association with inactivation of PKCε and Akt.\",\n      \"method\": \"GUCY1B3 overexpression and siRNA silencing in neonatal rat ventricular myocytes; in vitro IR model; specific PKCδ, PKCε, and Akt inhibitors; mouse coronary artery ligation model with GUCY1B3 knockdown; echocardiography, apoptosis assays\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — both in vitro and in vivo gain/loss-of-function with pharmacological pathway dissection, single lab study\",\n      \"pmids\": [\"30485489\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"GUCY1B3 (β subunit of sGC) participates in mediating cGMP production upstream of VEGF expression in glioma cells; antisense knockdown of GUCY1B3 in human glioma cell lines (CCF-STTG1 and U-87MG) markedly reduced cGMP content, VEGF expression, and in vitro angiogenic activity (HUVEC growth induction), and dramatically suppressed subcutaneous tumor formation and vascular index in nude mice.\",\n      \"method\": \"Antisense RNA transfection in glioma cell lines; cGMP measurement; VEGF expression analysis; HUVEC proliferation assay; xenograft tumor formation assay with vascular index measurement\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with multiple in vitro and in vivo readouts in a single lab study\",\n      \"pmids\": [\"15201957\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Co-expression of GUCY1A3 and GUCY1B3 subunits via lentiviral vectors in HEK293 cells resulted in formation of functional sGC enzyme with increased catalytic activity and elevated cellular cGMP levels, demonstrating that co-expression of the two subunits is sufficient for assembly of a functional heterodimeric enzyme.\",\n      \"method\": \"Lentiviral overexpression of GUCY1A3 and GUCY1B3 in HEK293 cells; sGC enzyme activity assay; cGMP quantification; cytotoxicity assay\",\n      \"journal\": \"Technology and health care\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro reconstitution of enzyme activity via co-expression, single lab, single study\",\n      \"pmids\": [\"36442224\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"GUCY1B1 is expressed in retinal vascular cells and neuronal elements (retinal ganglion, bipolar, and amacrine cells) in healthy human and rodent retinas; sGC function (requiring GUCY1B1) is impaired by oxidative stress in the retina, and the sGC activator runcaciguat activates sGC in multiple retinal cell types, counteracting oxidative stress-induced damage and improving neuroretinal function and morphology in rat retinal ischemia-reperfusion and streptozotocin-diabetic models.\",\n      \"method\": \"Immunohistochemistry for sGC subunit localization; in vitro and in vivo sGC activity assays; electroretinography; optokinetic tracking; retinal morphology assessment in rat IR and STZ-diabetic models\",\n      \"journal\": \"Diabetes\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiments combined with functional in vivo and in vitro assays, multiple orthogonal methods in one study\",\n      \"pmids\": [\"40249725\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In zebrafish, combined morpholino downregulation of both gucy1a1 and gucy1b1 (but not individual knockdown of either alone) increased blood flow and linear velocity, indicating that both subunits together are required for sGC-mediated regulation of vascular blood flow. An sGC stimulator (BAY41-2272) rescued impaired cGMP production in gucy1a1 mutant larvae.\",\n      \"method\": \"CRISPR-Cas9 gucy1a1 mutant zebrafish; individual and combined morpholino knockdown of gucy1a1 and gucy1b1; blood flow parameter measurement; pharmacological sGC stimulation; cGMP assay\",\n      \"journal\": \"Frontiers in physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and morpholino loss-of-function with functional blood flow readouts, multiple genetic conditions tested in single lab study\",\n      \"pmids\": [\"33716783\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In LPS-stimulated alveolar epithelial cells (rat RLE-6TN), ASIC1a negatively regulates LncRNA00178, which in turn acts as a sponge for miR-466b-3p; miR-466b-3p targets Gucy1b1 mRNA for degradation. ASIC1a thus suppresses Gucy1b1 expression via the LncRNA00178/miR-466b-3p axis, promoting apoptosis of alveolar epithelial cells in acute lung injury.\",\n      \"method\": \"High-throughput transcriptomic sequencing of rat lung tissue; plasmid transfection to alter ASIC1a, LncRNA00178, miR-466b-3p, Gucy1b1 expression in RLE-6TN cells; dual-luciferase reporter assay; apoptosis assays\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual-luciferase validation of miRNA-target interaction and direct manipulation of pathway components in a single lab study; note that this involves non-coding RNA regulation of the canonical protein\",\n      \"pmids\": [\"41830717\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"GUCY1B1 (alias GUCY1B3) encodes the β1 subunit of soluble guanylyl cyclase (sGC), an obligate heterodimer with the α1 subunit (GUCY1A1/GUCY1A3) that functions as the main intracellular receptor for nitric oxide (NO); upon NO binding, the heterodimer catalyzes cGMP synthesis to mediate vasodilation, cardiac protection (via PKCε/Akt), gastrointestinal smooth muscle relaxation (through SMCs and ICCs jointly), and retinal neuroprotection—with its transcription positively regulated by FoxO4 and Notch/MAML2/FRYL signaling and negatively regulated by hypertension-induced Notch pathway suppression and miR-466b-3p targeting.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"GUCY1B1 (β1 subunit, alias GUCY1B3) encodes one of the two obligate subunits of soluble guanylyl cyclase (sGC), the nitric oxide (NO) receptor that synthesizes cGMP; co-expression of GUCY1B1 with the α1 subunit (GUCY1A3) in heterologous cells is sufficient to assemble a catalytically active, cGMP-producing heterodimer [#0, #8]. Functional output requires both subunits in concert: in vivo, only combined loss of α1 and β1 abolishes sGC-dependent vascular and smooth-muscle responses, whereas knockdown of either alone is insufficient [#10]. Through this NO/sGC/cGMP axis, GUCY1B1 mediates a range of tissue-level processes—nitrergic gastrointestinal smooth muscle relaxation, in which it operates jointly in smooth muscle cells and interstitial cells of Cajal so that both populations are required for complete relaxation and normal gut transit [#2]; cardioprotection against ischemia-reperfusion injury via downstream PKCε/Akt signaling [#6]; and retinal neuroprotection against oxidative stress in vascular and neuronal cells [#9]. Downstream signaling proceeds through cGMP-dependent protein kinase (PKG) and its substrate VASP [#3, #5]. GUCY1B1 transcription is positively controlled by FoxO4, which binds FoxO motifs in the GUCY1B3 promoter, and by Notch signaling acting through coactivators MAML2 and FRYL, while hypertension represses sGC by suppressing this Notch pathway [#4, #5]; post-transcriptionally, Gucy1b1 mRNA is targeted for degradation by miR-466b-3p [#11]. In tumor and developmental settings the same axis is co-opted, contributing to glioma angiogenesis via cGMP-dependent VEGF expression [#7] and to Notch-driven endothelial-to-mesenchymal transition during cardiac valve development [#1].\",\n  \"teleology\": [\n    {\n      \"year\": 1990,\n      \"claim\": \"Established that GUCY1B1 encodes a distinct ~70-kDa β subunit of the soluble guanylyl cyclase heterodimer with tissue-selective expression, defining the molecular identity of the gene.\",\n      \"evidence\": \"PCR-based cDNA cloning and tissue mRNA expression profiling\",\n      \"pmids\": [\"1980215\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not demonstrate enzymatic activity of the reconstituted heterodimer\", \"Subunit stoichiometry and NO-binding role not yet assigned\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Linked GUCY1B1-dependent cGMP production to a pro-angiogenic program, showing the axis can drive VEGF expression and tumor vascularization in glioma.\",\n      \"evidence\": \"Antisense knockdown in human glioma cell lines with cGMP, VEGF, HUVEC angiogenesis, and xenograft readouts\",\n      \"pmids\": [\"15201957\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking cGMP to VEGF transcription not resolved\", \"Single-gene antisense without rescue\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Placed GUCY1B1 within a Notch-driven autocrine NO/sGC loop required for endothelial-to-mesenchymal transition in valve development, extending its role beyond mature smooth muscle.\",\n      \"evidence\": \"Genetic/pharmacological Notch manipulation in vivo and in vitro with EndMT functional assays\",\n      \"pmids\": [\"21839921\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct transcriptional control of GUCY1B1 promoter by Notch not demonstrated here\", \"Relative contribution of β1 vs α1 not separated\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined the cellular basis of nitrergic gastrointestinal relaxation, showing GUCY1B1 acts jointly in smooth muscle cells and interstitial cells of Cajal so that both are required for complete NO-induced relaxation.\",\n      \"evidence\": \"Cell-specific conditional knockout of GUCY1B3 in SMCs, ICCs, or both, with isometric force and gut transit measurements\",\n      \"pmids\": [\"23528627\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"cGMP effector steps downstream in each cell type not dissected\", \"Does not address other smooth muscle beds\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Connected Notch signaling to GUCY1B1 transcription and downstream PKG/VASP output, positioning Notch as a positive upstream regulator of the NO/sGC pathway.\",\n      \"evidence\": \"Notch3 activation and DAPT inhibition in ovarian epithelial/cancer cells with cGMP, PKG, and VASP phosphorylation readouts\",\n      \"pmids\": [\"24041655\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct promoter occupancy by Notch effectors shown\", \"Whether effect is direct or indirect unresolved\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identified the Notch coactivators MAML2 and FRYL as controllers of sGC expression and showed hypertension represses GUCY1B1 by suppressing this Notch axis, curtailing NO-dependent vasorelaxation.\",\n      \"evidence\": \"Angiotensin II hypertension model with gain/loss of Notch signaling, protein expression, and vasorelaxation assays across mouse, rat, and human data\",\n      \"pmids\": [\"28465505\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct binding of MAML2/FRYL complex to the GUCY1B1 promoter not shown\", \"Causality of repression in human hypertension inferred from expression\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established a cardioprotective function for GUCY1B1 against ischemia-reperfusion injury and traced it to PKCε/Akt signaling.\",\n      \"evidence\": \"Overexpression and siRNA silencing in neonatal rat ventricular myocytes plus mouse coronary ligation, with pharmacological PKC/Akt inhibitors and echocardiography\",\n      \"pmids\": [\"30485489\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How sGC/cGMP couples to PKCε/Akt activation mechanistically not defined\", \"Single lab; α1 subunit involvement not addressed\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated genetically that both subunits together are required for sGC-mediated vascular blood flow regulation, confirming obligate heterodimer function in vivo.\",\n      \"evidence\": \"CRISPR mutant and combined/individual morpholino knockdown of gucy1a1 and gucy1b1 in zebrafish with blood flow measurement and pharmacological sGC stimulation rescue\",\n      \"pmids\": [\"33716783\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Morpholino off-target effects inherent to method\", \"cGMP measurement performed in α1 mutant, not β1-specific\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified FoxO4 as a direct transcriptional activator of GUCY1B1 in vascular smooth muscle, with opposing roles for FoxO1/FoxO3, establishing isoform-specific transcriptional control of the gene.\",\n      \"evidence\": \"shRNA knockdown of FoxO family members in aortic SMCs with qRT-PCR, cGMP, promoter luciferase, and ChIP\",\n      \"pmids\": [\"35089807\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of opposing FoxO1/FoxO3 effect not resolved\", \"Interplay with Notch regulation not integrated\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Confirmed by reconstitution that co-expression of α1 and β1 subunits alone is sufficient to form a catalytically active cGMP-producing enzyme.\",\n      \"evidence\": \"Lentiviral co-expression of GUCY1A3 and GUCY1B3 in HEK293 cells with sGC activity and cGMP assays\",\n      \"pmids\": [\"36442224\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural or stoichiometric characterization\", \"NO-responsiveness of the reconstituted enzyme not tested here\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Extended GUCY1B1 function to retinal neuroprotection, localizing sGC to retinal vascular and neuronal cells and showing pharmacological sGC activation counteracts oxidative-stress damage.\",\n      \"evidence\": \"Immunohistochemistry plus in vitro/in vivo sGC activity assays and functional retinal readouts in rat IR and STZ-diabetic models\",\n      \"pmids\": [\"40249725\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"β1-specific genetic loss-of-function in retina not performed\", \"Downstream cGMP effectors in neurons not identified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Revealed post-transcriptional control of Gucy1b1 by a non-coding RNA network in alveolar epithelial injury, with miR-466b-3p directly targeting its mRNA.\",\n      \"evidence\": \"Transcriptomic sequencing, pathway component manipulation, and dual-luciferase reporter assay in LPS-stimulated rat alveolar epithelial cells\",\n      \"pmids\": [\"41830717\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"sGC enzymatic/cGMP consequence of Gucy1b1 repression not measured\", \"Single lab; ASIC1a/lncRNA axis not validated in vivo\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the multiple converging transcriptional (FoxO4, Notch/MAML2/FRYL) and post-transcriptional (miR-466b-3p) regulators are integrated to set GUCY1B1 levels across tissues, and the structural basis of NO sensing by the β1 subunit, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model linking the distinct regulatory inputs\", \"No structural characterization of the β1 NO-binding module in the timeline\", \"Tissue-specific effector pathways downstream of cGMP only partially mapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0009975\", \"supporting_discovery_ids\": [0, 8]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 8, 10]}\n    ],\n    \"complexes\": [\"soluble guanylyl cyclase (sGC) heterodimer\"],\n    \"partners\": [\"GUCY1A3\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}