{"gene":"CNN1","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2016,"finding":"CNN1 (calponin 1) is an actin filament-associated protein that inhibits actin-activated myosin ATPase activity and stabilizes the actin cytoskeleton; it is specifically expressed in smooth muscle cells and fine-tunes smooth muscle contractility. Phosphorylation regulates calponin function.","method":"Review synthesizing three decades of biochemical assays (actin-activated myosin ATPase assays, actin-binding experiments, phosphorylation studies) and cell-type expression analyses","journal":"Gene","confidence":"High","confidence_rationale":"Tier 1 / Strong — multiple independent labs over decades using in vitro enzymatic assays, reconstitution, and mutagenesis, consolidated in review","pmids":["26970176"],"is_preprint":false},{"year":2020,"finding":"miR-106b-5p directly binds the 3'UTR of CNN1 mRNA (confirmed by dual-luciferase assay), suppressing CNN1 expression and activating the Rho/ROCK1 signaling pathway, thereby promoting breast cancer cell migration, invasion, and lung metastasis in vivo.","method":"Dual-luciferase reporter assay, transcript half-life assay, CNN1 overexpression/knockdown in cell lines, Rho/ROCK1 inhibition assay, in vivo lung metastasis assay","journal":"Aging","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dual-luciferase plus in vivo rescue, single lab with multiple orthogonal methods","pmids":["31986487"],"is_preprint":false},{"year":2020,"finding":"The lncRNA LINC00337 recruits DNMT1 to the CNN1 promoter region, causing hypermethylation and transcriptional suppression of CNN1 in colorectal cancer; CNN1 suppression promotes tumor angiogenesis (elevated VEGF, microvascular density) and cell proliferation/invasion.","method":"Bisulfite sequencing PCR (BSP), methylation-specific PCR (MSP), loss- and gain-of-function experiments in vitro and in nude mouse xenograft models, tube formation assay","journal":"Cancer gene therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — chromatin recruitment assay (DNMT1), bisulfite sequencing, in vivo xenograft, single lab with multiple orthogonal methods","pmids":["33328585"],"is_preprint":false},{"year":2021,"finding":"CNN1 overexpression in lung squamous cell carcinoma cells activates TIMP2 expression, which in turn upregulates DKK1 to inhibit the Wnt/β-catenin/c-myc signaling pathway, resulting in suppressed invasion, migration, and epithelial-to-mesenchymal transition (reduced MMP2, MMP9, N-cadherin, SLUG; preserved E-cadherin).","method":"Transient transfection overexpression, wound healing assay, Transwell invasion assay, RT-qPCR, western blotting, immunofluorescence","journal":"Experimental and therapeutic medicine","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, overexpression only, no direct TIMP2-CNN1 binding demonstrated, pathway placement inferred from expression changes","pmids":["34178128"],"is_preprint":false},{"year":2022,"finding":"CNN1 overexpression in bladder cancer cells decreases protein levels of HIF-1α and its downstream glycolysis effectors (GLUT1, PKM2, LDHA, PDK1), suppressing glucose absorption, lactate secretion, ATP production, and inhibiting tumor proliferation and invasion in vitro and in vivo.","method":"CNN1 overexpression in bladder cancer cells, CCK8 proliferation assay, Transwell invasion/migration assay, metabolic flux assays (glucose, lactate, ATP), western blotting, nude mouse xenograft with IHC","journal":"Frontiers in oncology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, overexpression with phenotypic readouts but no direct biochemical mechanism linking CNN1 to HIF-1α established","pmids":["35903683"],"is_preprint":false},{"year":2024,"finding":"Kdm6a (UTX) forms an epigenetic regulatory axis by binding to CNN1, orchestrating trauma-induced spinal cord microvascular endothelial cell (SCMEC) senescence and SASP secretion; UTX deletion in endothelial cells protects against senescence, reduces proinflammatory SASP factors, and promotes neurological recovery after spinal cord injury.","method":"Endothelial cell-specific UTX knockout mouse model, ChIP/binding assays demonstrating UTX–CNN1 interaction, SASP cytokine profiling, neurological functional recovery assessment, senolytic drug delivery experiments","journal":"Bone research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo genetic KO with defined cellular phenotype and binding evidence, single lab with multiple orthogonal approaches","pmids":["38528029"],"is_preprint":false},{"year":2025,"finding":"USP48, a ubiquitin-specific protease, directly stabilizes CNN1 protein (preventing its degradation); the USP48/CNN1 axis inhibits ERK1/2 phosphorylation and thereby suppresses CXCL1 and CXCL2 release, conferring cardioprotection against diabetes-aggravated myocardial ischemia/reperfusion injury.","method":"Proteomics, cardiac-specific USP48 KO and overexpression in mice, CNN1 silencing rescue experiments, RNA sequencing, western blotting for ERK1/2 activation, CXCL1/2 neutralizing antibody blockade","journal":"Metabolism: clinical and experimental","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo genetic models (KO and OE) with proteomics, silencing rescue, and pathway validation; single lab","pmids":["40513941"],"is_preprint":false},{"year":2024,"finding":"CNN1 knockdown in chicken breast muscle satellite cells reduces proliferation and differentiation and decreases lipid accumulation during differentiation, demonstrating a functional role for CNN1 in satellite cell-mediated skeletal muscle growth.","method":"siRNA-mediated CNN1 knockdown in primary satellite cells, proliferation and differentiation assays, lipid accumulation assays","journal":"Poultry science","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single knockdown approach in non-mammalian (chicken) cells, no molecular mechanism defined","pmids":["38669818"],"is_preprint":false},{"year":2024,"finding":"lncRNA MEG3 acts as a competing endogenous RNA (ceRNA) by directly binding miR-330 (confirmed by dual-luciferase and RIP assays), preventing miR-330-mediated suppression of CNN1; restoration of miR-330 on top of MEG3 overexpression reduces CNN1 expression and reverses the tumor-suppressive effects of MEG3 in breast cancer cells.","method":"Dual-luciferase reporter assay, RNA immunoprecipitation (RIP), miRNA mimic transfection rescue experiments, cell viability, migration, invasion, and apoptosis assays","journal":"Aging","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dual-luciferase plus RIP confirm direct MEG3–miR-330 interaction, rescue experiments support CNN1 as downstream effector; single lab","pmids":["38240701"],"is_preprint":false},{"year":2025,"finding":"CNN1 overexpression activates p53 signaling, which in turn directly binds the SLC7A11 promoter to suppress its transcription; reduced SLC7A11 induces ferroptosis in colorectal cancer cells.","method":"CNN1 overexpression (UTMD-assisted transfection), p53 pathway reporter assays, promoter-binding analysis for p53–SLC7A11, ferroptosis assays, rescue experiments","journal":"Biochemical and biophysical research communications","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, no direct CNN1–p53 biochemical interaction demonstrated; pathway placement inferred from expression and rescue experiments","pmids":["40602187"],"is_preprint":false},{"year":2026,"finding":"Histone lactylation (H3K18la, H4K12la) upregulates CNN1 expression in castration-resistant prostate cancer; elevated CNN1 promotes autophagy and G0/G1 cell cycle arrest, contributing to docetaxel resistance. Inhibition of CNN1 reverses these effects, and in vivo CNN1 overexpression restores tumor growth under lactate inhibition.","method":"Clinical CRPC tissue analysis, lactate inhibition (2-DG/oxamate), CNN1 OE/KO in CRPC cell lines and xenograft mouse models, cell cycle analysis, autophagy assays","journal":"Cell death discovery","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, epigenetic–CNN1 link inferred from correlative histone mark data, no direct histone lactylation writer-CNN1 promoter interaction shown","pmids":["42129130"],"is_preprint":false}],"current_model":"CNN1 (calponin 1) is a smooth muscle-specific actin filament-associated protein that inhibits actin-activated myosin ATPase and stabilizes the actin cytoskeleton to fine-tune contractility; its activity is regulated by phosphorylation, by miRNA-mediated mRNA suppression (e.g., miR-106b-5p, miR-330), by promoter methylation via DNMT1 recruitment (through lncRNA LINC00337), by histone lactylation, by ubiquitin-mediated protein stability (USP48), and by the epigenetic regulator UTX/Kdm6a; downstream, CNN1 modulates Rho/ROCK1, HIF-1α/glycolysis, DKK1/Wnt/β-catenin, ERK1/2/CXCL1-2, and p53/SLC7A11 pathways to suppress cell migration, invasion, metabolic reprogramming, and inflammation in multiple cell contexts."},"narrative":{"mechanistic_narrative":"CNN1 (calponin 1) is a smooth muscle-specific actin filament-associated protein that inhibits actin-activated myosin ATPase activity and stabilizes the actin cytoskeleton to fine-tune smooth muscle contractility, with its activity modulated by phosphorylation [PMID:26970176]. Beyond this contractile role, the available corpus characterizes CNN1 predominantly as a downstream effector whose expression is set by multiple regulatory layers and whose restoration suppresses malignant and pathological cell behaviors. CNN1 abundance is constrained transcriptionally by lncRNA LINC00337, which recruits DNMT1 to hypermethylate the CNN1 promoter [PMID:33328585], and post-transcriptionally by miRNAs targeting its 3'UTR, including miR-106b-5p [PMID:31986487] and miR-330, the latter sequestered by the ceRNA lncRNA MEG3 [PMID:38240701]; CNN1 protein is further stabilized against degradation by the deubiquitinase USP48 [PMID:40513941]. When restored, CNN1 acts through several effector pathways: suppression of Rho/ROCK1 signaling to limit migration and invasion [PMID:31986487], and inhibition of ERK1/2 phosphorylation to reduce CXCL1/CXCL2 release in a cardioprotective context [PMID:40513941]. CNN1 also participates in an epigenetic axis with Kdm6a/UTX governing endothelial senescence and SASP secretion after spinal cord injury [PMID:38528029].","teleology":[{"year":2016,"claim":"Established the core biochemical identity of CNN1 as a smooth-muscle actin-binding protein that restrains myosin ATPase and is phosphoregulated, defining its baseline contractile function.","evidence":"Review consolidating decades of in vitro actin-activated myosin ATPase assays, actin-binding and phosphorylation studies, and expression analyses","pmids":["26970176"],"confidence":"High","gaps":["Does not address CNN1 function outside smooth muscle","Does not connect contractile biochemistry to transcriptional/cancer regulation"]},{"year":2020,"claim":"Showed CNN1 is a direct miRNA target whose loss activates a pro-metastatic signaling cascade, framing CNN1 as a suppressor of migration/invasion.","evidence":"Dual-luciferase 3'UTR assay, overexpression/knockdown, Rho/ROCK1 inhibition, and in vivo lung metastasis assay in breast cancer cells","pmids":["31986487"],"confidence":"Medium","gaps":["Direct CNN1-Rho/ROCK1 biochemical link not shown","miR-106b-5p effects may involve other targets"]},{"year":2020,"claim":"Identified an epigenetic mechanism (lncRNA-directed DNMT1 promoter methylation) that silences CNN1, linking its loss to tumor angiogenesis and proliferation.","evidence":"Bisulfite/methylation-specific PCR, DNMT1 recruitment assays, and xenograft/tube formation assays in colorectal cancer","pmids":["33328585"],"confidence":"Medium","gaps":["Mechanism linking CNN1 loss to VEGF induction not defined","Single lab"]},{"year":2021,"claim":"Proposed that restored CNN1 suppresses EMT and Wnt/β-catenin signaling via a TIMP2/DKK1 axis, extending its tumor-suppressive reach.","evidence":"Overexpression with wound-healing, Transwell, RT-qPCR and western blot in lung squamous carcinoma","pmids":["34178128"],"confidence":"Low","gaps":["No direct TIMP2-CNN1 binding demonstrated; pathway placement inferred from expression changes","Overexpression-only design"]},{"year":2022,"claim":"Linked CNN1 to metabolic reprogramming by showing its overexpression downregulates HIF-1α-driven glycolysis in bladder cancer.","evidence":"Overexpression with metabolic flux assays, proliferation/invasion assays, and xenografts","pmids":["35903683"],"confidence":"Low","gaps":["No biochemical mechanism connecting CNN1 to HIF-1α established","Single lab, overexpression-driven"]},{"year":2024,"claim":"Placed CNN1 within an epigenetic axis with Kdm6a/UTX controlling endothelial senescence, broadening its role beyond cancer into injury/inflammation biology.","evidence":"Endothelial-specific UTX knockout mice, UTX-CNN1 binding/ChIP assays, SASP profiling, and functional recovery after spinal cord injury","pmids":["38528029"],"confidence":"Medium","gaps":["Directionality of UTX effect on CNN1 expression not fully resolved","Functional contribution of CNN1 itself to senescence not isolated from UTX"]},{"year":2024,"claim":"Demonstrated a non-smooth-muscle role for CNN1 in satellite-cell proliferation, differentiation, and lipid accumulation during myogenesis.","evidence":"siRNA knockdown in primary chicken breast muscle satellite cells with proliferation/differentiation/lipid assays","pmids":["38669818"],"confidence":"Low","gaps":["No molecular mechanism defined","Single knockdown approach in non-mammalian cells"]},{"year":2024,"claim":"Confirmed direct MEG3-miR-330 interaction that relieves miRNA suppression of CNN1, reinforcing a ceRNA layer controlling CNN1 levels.","evidence":"Dual-luciferase and RIP assays plus miRNA-mimic rescue in breast cancer cells","pmids":["38240701"],"confidence":"Medium","gaps":["CNN1 downstream effectors in this context not mapped","Single lab"]},{"year":2025,"claim":"Identified USP48 as a deubiquitinase that stabilizes CNN1 protein, defining a post-translational control point with cardioprotective consequences via ERK1/2-CXCL1/2 suppression.","evidence":"Proteomics, cardiac-specific USP48 KO/OE mice, CNN1-silencing rescue, RNA-seq, and CXCL1/2 neutralization","pmids":["40513941"],"confidence":"Medium","gaps":["Direct USP48-CNN1 deubiquitination biochemistry not fully reconstituted","Mechanism by which CNN1 represses ERK1/2 unclear"]},{"year":2025,"claim":"Connected CNN1 to ferroptosis induction through a p53/SLC7A11 axis in colorectal cancer.","evidence":"CNN1 overexpression, p53 reporter and SLC7A11 promoter-binding assays, ferroptosis and rescue assays","pmids":["40602187"],"confidence":"Low","gaps":["No direct CNN1-p53 biochemical interaction shown","Pathway placement inferred from expression/rescue data"]},{"year":2026,"claim":"Reported that histone lactylation upregulates CNN1 to drive autophagy, cell-cycle arrest, and docetaxel resistance in castration-resistant prostate cancer, casting CNN1 context-dependently as pro-survival.","evidence":"CRPC tissue analysis, lactate inhibition, CNN1 OE/KO in cell lines and xenografts, cell cycle and autophagy assays","pmids":["42129130"],"confidence":"Low","gaps":["Epigenetic-CNN1 link correlative; no direct lactylation writer-promoter interaction shown","Mechanism linking CNN1 to autophagy/arrest undefined"]},{"year":null,"claim":"How CNN1's defined actin/myosin biochemistry mechanistically couples to the diverse signaling pathways (Rho/ROCK1, ERK1/2, HIF-1α, p53/SLC7A11, Wnt) attributed to it remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No direct biochemical link established between CNN1's cytoskeletal activity and downstream signaling effectors","Most non-muscle pathway claims rest on single-lab overexpression/correlative data"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-397014","term_label":"Muscle contraction","supporting_discovery_ids":[0]}],"complexes":[],"partners":["USP48","KDM6A"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P51911","full_name":"Calponin-1","aliases":["Basic calponin","Calponin H1, smooth muscle"],"length_aa":297,"mass_kda":33.2,"function":"Thin filament-associated protein that is implicated in the regulation and modulation of smooth muscle contraction. It is capable of binding to actin, calmodulin and tropomyosin. The interaction of calponin with actin inhibits the actomyosin Mg-ATPase activity (By similarity)","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/P51911/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CNN1","classification":"Not Classified","n_dependent_lines":8,"n_total_lines":1208,"dependency_fraction":0.006622516556291391},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CNN1","total_profiled":1310},"omim":[{"mim_id":"611795","title":"MICRO RNA 145; MIR145","url":"https://www.omim.org/entry/611795"},{"mim_id":"610674","title":"SPERM FLAGELLAR PROTEIN 1; SPEF1","url":"https://www.omim.org/entry/610674"},{"mim_id":"610077","title":"REGULATOR OF CELL CYCLE; RGCC","url":"https://www.omim.org/entry/610077"},{"mim_id":"609922","title":"EH DOMAIN-BINDING PROTEIN 1; EHBP1","url":"https://www.omim.org/entry/609922"},{"mim_id":"608667","title":"NIPPED-B-LIKE; NIPBL","url":"https://www.omim.org/entry/608667"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Actin filaments","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"blood vessel","ntpm":2087.4},{"tissue":"intestine","ntpm":2743.0},{"tissue":"smooth muscle","ntpm":2059.2}],"url":"https://www.proteinatlas.org/search/CNN1"},"hgnc":{"alias_symbol":["SMCC","Sm-Calp"],"prev_symbol":[]},"alphafold":{"accession":"P51911","domains":[{"cath_id":"1.10.418.10","chopping":"12-133","consensus_level":"high","plddt":90.9106,"start":12,"end":133}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P51911","model_url":"https://alphafold.ebi.ac.uk/files/AF-P51911-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P51911-F1-predicted_aligned_error_v6.png","plddt_mean":71.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CNN1","jax_strain_url":"https://www.jax.org/strain/search?query=CNN1"},"sequence":{"accession":"P51911","fasta_url":"https://rest.uniprot.org/uniprotkb/P51911.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P51911/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P51911"}},"corpus_meta":[{"pmid":"26970176","id":"PMC_26970176","title":"Calponin isoforms CNN1, CNN2 and CNN3: Regulators for actin cytoskeleton functions in smooth muscle and non-muscle cells.","date":"2016","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/26970176","citation_count":150,"is_preprint":false},{"pmid":"22561345","id":"PMC_22561345","title":"Cnn1 inhibits the interactions between the KMN complexes of the yeast kinetochore.","date":"2012","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/22561345","citation_count":90,"is_preprint":false},{"pmid":"31986487","id":"PMC_31986487","title":"miR-106b-5p contributes to the lung metastasis of breast cancer via targeting CNN1 and regulating Rho/ROCK1 pathway.","date":"2020","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/31986487","citation_count":73,"is_preprint":false},{"pmid":"33328585","id":"PMC_33328585","title":"LINC00337 promotes tumor angiogenesis in colorectal cancer by recruiting DNMT1, which suppresses the expression of CNN1.","date":"2020","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/33328585","citation_count":29,"is_preprint":false},{"pmid":"34178128","id":"PMC_34178128","title":"CNN1 regulates the DKK1/Wnt/β-catenin/c-myc signaling pathway by activating TIMP2 to inhibit the invasion, migration and EMT of lung squamous cell carcinoma cells.","date":"2021","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34178128","citation_count":22,"is_preprint":false},{"pmid":"32679099","id":"PMC_32679099","title":"The Structural Basis for Kinetochore Stabilization by Cnn1/CENP-T.","date":"2020","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/32679099","citation_count":16,"is_preprint":false},{"pmid":"35903683","id":"PMC_35903683","title":"CNN1 Represses Bladder Cancer Progression and Metabolic Reprogramming by Modulating HIF-1α Signaling Pathway.","date":"2022","source":"Frontiers in 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miR-330/CNN1.","date":"2024","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/38240701","citation_count":4,"is_preprint":false},{"pmid":"40513941","id":"PMC_40513941","title":"USP48 protects against myocardial ischemia-reperfusion injury by stabilizing and upregulating CNN1 in type 1 diabetes mice.","date":"2025","source":"Metabolism: clinical and experimental","url":"https://pubmed.ncbi.nlm.nih.gov/40513941","citation_count":2,"is_preprint":false},{"pmid":"40602187","id":"PMC_40602187","title":"Ultrasound-targeted microbubble destruction mediated upregulation of CNN1 induces ferroptosis in colorectal cancer cells by regulating p53-related SLC7A11 expression.","date":"2025","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/40602187","citation_count":1,"is_preprint":false},{"pmid":"42129130","id":"PMC_42129130","title":"Histone lactylation modification promotes docetaxel resistance and tumor progression through CNN1-Mediated 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Phosphorylation regulates calponin function.\",\n      \"method\": \"Review synthesizing three decades of biochemical assays (actin-activated myosin ATPase assays, actin-binding experiments, phosphorylation studies) and cell-type expression analyses\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — multiple independent labs over decades using in vitro enzymatic assays, reconstitution, and mutagenesis, consolidated in review\",\n      \"pmids\": [\"26970176\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"miR-106b-5p directly binds the 3'UTR of CNN1 mRNA (confirmed by dual-luciferase assay), suppressing CNN1 expression and activating the Rho/ROCK1 signaling pathway, thereby promoting breast cancer cell migration, invasion, and lung metastasis in vivo.\",\n      \"method\": \"Dual-luciferase reporter assay, transcript half-life assay, CNN1 overexpression/knockdown in cell lines, Rho/ROCK1 inhibition assay, in vivo lung metastasis assay\",\n      \"journal\": \"Aging\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual-luciferase plus in vivo rescue, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"31986487\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"The lncRNA LINC00337 recruits DNMT1 to the CNN1 promoter region, causing hypermethylation and transcriptional suppression of CNN1 in colorectal cancer; CNN1 suppression promotes tumor angiogenesis (elevated VEGF, microvascular density) and cell proliferation/invasion.\",\n      \"method\": \"Bisulfite sequencing PCR (BSP), methylation-specific PCR (MSP), loss- and gain-of-function experiments in vitro and in nude mouse xenograft models, tube formation assay\",\n      \"journal\": \"Cancer gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — chromatin recruitment assay (DNMT1), bisulfite sequencing, in vivo xenograft, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"33328585\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CNN1 overexpression in lung squamous cell carcinoma cells activates TIMP2 expression, which in turn upregulates DKK1 to inhibit the Wnt/β-catenin/c-myc signaling pathway, resulting in suppressed invasion, migration, and epithelial-to-mesenchymal transition (reduced MMP2, MMP9, N-cadherin, SLUG; preserved E-cadherin).\",\n      \"method\": \"Transient transfection overexpression, wound healing assay, Transwell invasion assay, RT-qPCR, western blotting, immunofluorescence\",\n      \"journal\": \"Experimental and therapeutic medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, overexpression only, no direct TIMP2-CNN1 binding demonstrated, pathway placement inferred from expression changes\",\n      \"pmids\": [\"34178128\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CNN1 overexpression in bladder cancer cells decreases protein levels of HIF-1α and its downstream glycolysis effectors (GLUT1, PKM2, LDHA, PDK1), suppressing glucose absorption, lactate secretion, ATP production, and inhibiting tumor proliferation and invasion in vitro and in vivo.\",\n      \"method\": \"CNN1 overexpression in bladder cancer cells, CCK8 proliferation assay, Transwell invasion/migration assay, metabolic flux assays (glucose, lactate, ATP), western blotting, nude mouse xenograft with IHC\",\n      \"journal\": \"Frontiers in oncology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, overexpression with phenotypic readouts but no direct biochemical mechanism linking CNN1 to HIF-1α established\",\n      \"pmids\": [\"35903683\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Kdm6a (UTX) forms an epigenetic regulatory axis by binding to CNN1, orchestrating trauma-induced spinal cord microvascular endothelial cell (SCMEC) senescence and SASP secretion; UTX deletion in endothelial cells protects against senescence, reduces proinflammatory SASP factors, and promotes neurological recovery after spinal cord injury.\",\n      \"method\": \"Endothelial cell-specific UTX knockout mouse model, ChIP/binding assays demonstrating UTX–CNN1 interaction, SASP cytokine profiling, neurological functional recovery assessment, senolytic drug delivery experiments\",\n      \"journal\": \"Bone research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic KO with defined cellular phenotype and binding evidence, single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"38528029\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"USP48, a ubiquitin-specific protease, directly stabilizes CNN1 protein (preventing its degradation); the USP48/CNN1 axis inhibits ERK1/2 phosphorylation and thereby suppresses CXCL1 and CXCL2 release, conferring cardioprotection against diabetes-aggravated myocardial ischemia/reperfusion injury.\",\n      \"method\": \"Proteomics, cardiac-specific USP48 KO and overexpression in mice, CNN1 silencing rescue experiments, RNA sequencing, western blotting for ERK1/2 activation, CXCL1/2 neutralizing antibody blockade\",\n      \"journal\": \"Metabolism: clinical and experimental\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic models (KO and OE) with proteomics, silencing rescue, and pathway validation; single lab\",\n      \"pmids\": [\"40513941\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CNN1 knockdown in chicken breast muscle satellite cells reduces proliferation and differentiation and decreases lipid accumulation during differentiation, demonstrating a functional role for CNN1 in satellite cell-mediated skeletal muscle growth.\",\n      \"method\": \"siRNA-mediated CNN1 knockdown in primary satellite cells, proliferation and differentiation assays, lipid accumulation assays\",\n      \"journal\": \"Poultry science\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single knockdown approach in non-mammalian (chicken) cells, no molecular mechanism defined\",\n      \"pmids\": [\"38669818\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"lncRNA MEG3 acts as a competing endogenous RNA (ceRNA) by directly binding miR-330 (confirmed by dual-luciferase and RIP assays), preventing miR-330-mediated suppression of CNN1; restoration of miR-330 on top of MEG3 overexpression reduces CNN1 expression and reverses the tumor-suppressive effects of MEG3 in breast cancer cells.\",\n      \"method\": \"Dual-luciferase reporter assay, RNA immunoprecipitation (RIP), miRNA mimic transfection rescue experiments, cell viability, migration, invasion, and apoptosis assays\",\n      \"journal\": \"Aging\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual-luciferase plus RIP confirm direct MEG3–miR-330 interaction, rescue experiments support CNN1 as downstream effector; single lab\",\n      \"pmids\": [\"38240701\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CNN1 overexpression activates p53 signaling, which in turn directly binds the SLC7A11 promoter to suppress its transcription; reduced SLC7A11 induces ferroptosis in colorectal cancer cells.\",\n      \"method\": \"CNN1 overexpression (UTMD-assisted transfection), p53 pathway reporter assays, promoter-binding analysis for p53–SLC7A11, ferroptosis assays, rescue experiments\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, no direct CNN1–p53 biochemical interaction demonstrated; pathway placement inferred from expression and rescue experiments\",\n      \"pmids\": [\"40602187\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Histone lactylation (H3K18la, H4K12la) upregulates CNN1 expression in castration-resistant prostate cancer; elevated CNN1 promotes autophagy and G0/G1 cell cycle arrest, contributing to docetaxel resistance. Inhibition of CNN1 reverses these effects, and in vivo CNN1 overexpression restores tumor growth under lactate inhibition.\",\n      \"method\": \"Clinical CRPC tissue analysis, lactate inhibition (2-DG/oxamate), CNN1 OE/KO in CRPC cell lines and xenograft mouse models, cell cycle analysis, autophagy assays\",\n      \"journal\": \"Cell death discovery\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, epigenetic–CNN1 link inferred from correlative histone mark data, no direct histone lactylation writer-CNN1 promoter interaction shown\",\n      \"pmids\": [\"42129130\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CNN1 (calponin 1) is a smooth muscle-specific actin filament-associated protein that inhibits actin-activated myosin ATPase and stabilizes the actin cytoskeleton to fine-tune contractility; its activity is regulated by phosphorylation, by miRNA-mediated mRNA suppression (e.g., miR-106b-5p, miR-330), by promoter methylation via DNMT1 recruitment (through lncRNA LINC00337), by histone lactylation, by ubiquitin-mediated protein stability (USP48), and by the epigenetic regulator UTX/Kdm6a; downstream, CNN1 modulates Rho/ROCK1, HIF-1α/glycolysis, DKK1/Wnt/β-catenin, ERK1/2/CXCL1-2, and p53/SLC7A11 pathways to suppress cell migration, invasion, metabolic reprogramming, and inflammation in multiple cell contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CNN1 (calponin 1) is a smooth muscle-specific actin filament-associated protein that inhibits actin-activated myosin ATPase activity and stabilizes the actin cytoskeleton to fine-tune smooth muscle contractility, with its activity modulated by phosphorylation [#0]. Beyond this contractile role, the available corpus characterizes CNN1 predominantly as a downstream effector whose expression is set by multiple regulatory layers and whose restoration suppresses malignant and pathological cell behaviors. CNN1 abundance is constrained transcriptionally by lncRNA LINC00337, which recruits DNMT1 to hypermethylate the CNN1 promoter [#2], and post-transcriptionally by miRNAs targeting its 3'UTR, including miR-106b-5p [#1] and miR-330, the latter sequestered by the ceRNA lncRNA MEG3 [#8]; CNN1 protein is further stabilized against degradation by the deubiquitinase USP48 [#6]. When restored, CNN1 acts through several effector pathways: suppression of Rho/ROCK1 signaling to limit migration and invasion [#1], and inhibition of ERK1/2 phosphorylation to reduce CXCL1/CXCL2 release in a cardioprotective context [#6]. CNN1 also participates in an epigenetic axis with Kdm6a/UTX governing endothelial senescence and SASP secretion after spinal cord injury [#5].\",\n  \"teleology\": [\n    {\n      \"year\": 2016,\n      \"claim\": \"Established the core biochemical identity of CNN1 as a smooth-muscle actin-binding protein that restrains myosin ATPase and is phosphoregulated, defining its baseline contractile function.\",\n      \"evidence\": \"Review consolidating decades of in vitro actin-activated myosin ATPase assays, actin-binding and phosphorylation studies, and expression analyses\",\n      \"pmids\": [\"26970176\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not address CNN1 function outside smooth muscle\", \"Does not connect contractile biochemistry to transcriptional/cancer regulation\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed CNN1 is a direct miRNA target whose loss activates a pro-metastatic signaling cascade, framing CNN1 as a suppressor of migration/invasion.\",\n      \"evidence\": \"Dual-luciferase 3'UTR assay, overexpression/knockdown, Rho/ROCK1 inhibition, and in vivo lung metastasis assay in breast cancer cells\",\n      \"pmids\": [\"31986487\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct CNN1-Rho/ROCK1 biochemical link not shown\", \"miR-106b-5p effects may involve other targets\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified an epigenetic mechanism (lncRNA-directed DNMT1 promoter methylation) that silences CNN1, linking its loss to tumor angiogenesis and proliferation.\",\n      \"evidence\": \"Bisulfite/methylation-specific PCR, DNMT1 recruitment assays, and xenograft/tube formation assays in colorectal cancer\",\n      \"pmids\": [\"33328585\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking CNN1 loss to VEGF induction not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Proposed that restored CNN1 suppresses EMT and Wnt/\\u03b2-catenin signaling via a TIMP2/DKK1 axis, extending its tumor-suppressive reach.\",\n      \"evidence\": \"Overexpression with wound-healing, Transwell, RT-qPCR and western blot in lung squamous carcinoma\",\n      \"pmids\": [\"34178128\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct TIMP2-CNN1 binding demonstrated; pathway placement inferred from expression changes\", \"Overexpression-only design\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Linked CNN1 to metabolic reprogramming by showing its overexpression downregulates HIF-1\\u03b1-driven glycolysis in bladder cancer.\",\n      \"evidence\": \"Overexpression with metabolic flux assays, proliferation/invasion assays, and xenografts\",\n      \"pmids\": [\"35903683\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No biochemical mechanism connecting CNN1 to HIF-1\\u03b1 established\", \"Single lab, overexpression-driven\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placed CNN1 within an epigenetic axis with Kdm6a/UTX controlling endothelial senescence, broadening its role beyond cancer into injury/inflammation biology.\",\n      \"evidence\": \"Endothelial-specific UTX knockout mice, UTX-CNN1 binding/ChIP assays, SASP profiling, and functional recovery after spinal cord injury\",\n      \"pmids\": [\"38528029\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Directionality of UTX effect on CNN1 expression not fully resolved\", \"Functional contribution of CNN1 itself to senescence not isolated from UTX\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrated a non-smooth-muscle role for CNN1 in satellite-cell proliferation, differentiation, and lipid accumulation during myogenesis.\",\n      \"evidence\": \"siRNA knockdown in primary chicken breast muscle satellite cells with proliferation/differentiation/lipid assays\",\n      \"pmids\": [\"38669818\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No molecular mechanism defined\", \"Single knockdown approach in non-mammalian cells\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Confirmed direct MEG3-miR-330 interaction that relieves miRNA suppression of CNN1, reinforcing a ceRNA layer controlling CNN1 levels.\",\n      \"evidence\": \"Dual-luciferase and RIP assays plus miRNA-mimic rescue in breast cancer cells\",\n      \"pmids\": [\"38240701\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"CNN1 downstream effectors in this context not mapped\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified USP48 as a deubiquitinase that stabilizes CNN1 protein, defining a post-translational control point with cardioprotective consequences via ERK1/2-CXCL1/2 suppression.\",\n      \"evidence\": \"Proteomics, cardiac-specific USP48 KO/OE mice, CNN1-silencing rescue, RNA-seq, and CXCL1/2 neutralization\",\n      \"pmids\": [\"40513941\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct USP48-CNN1 deubiquitination biochemistry not fully reconstituted\", \"Mechanism by which CNN1 represses ERK1/2 unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected CNN1 to ferroptosis induction through a p53/SLC7A11 axis in colorectal cancer.\",\n      \"evidence\": \"CNN1 overexpression, p53 reporter and SLC7A11 promoter-binding assays, ferroptosis and rescue assays\",\n      \"pmids\": [\"40602187\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct CNN1-p53 biochemical interaction shown\", \"Pathway placement inferred from expression/rescue data\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Reported that histone lactylation upregulates CNN1 to drive autophagy, cell-cycle arrest, and docetaxel resistance in castration-resistant prostate cancer, casting CNN1 context-dependently as pro-survival.\",\n      \"evidence\": \"CRPC tissue analysis, lactate inhibition, CNN1 OE/KO in cell lines and xenografts, cell cycle and autophagy assays\",\n      \"pmids\": [\"42129130\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Epigenetic-CNN1 link correlative; no direct lactylation writer-promoter interaction shown\", \"Mechanism linking CNN1 to autophagy/arrest undefined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How CNN1's defined actin/myosin biochemistry mechanistically couples to the diverse signaling pathways (Rho/ROCK1, ERK1/2, HIF-1\\u03b1, p53/SLC7A11, Wnt) attributed to it remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct biochemical link established between CNN1's cytoskeletal activity and downstream signaling effectors\", \"Most non-muscle pathway claims rest on single-lab overexpression/correlative data\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-397014\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"USP48\", \"KDM6A\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":4,"faith_pct":100.0}}