Pub Date : 2025-12-05Epub Date: 2025-10-30DOI: 10.1161/CIRCRESAHA.125.326480
Chang-Ru Tsai, Lin Liu, Yi Zhao, Jong H Kim, Paulo Czarnewski, Rich Gang Li, Fansen Meng, Mingjie Zheng, Jeffrey Steimle, Xiaolei Zhao, Francisco Grisanti, Zheng Sun, Jun Wang, Md Abul Hassan Samee, Xiao Li, James F Martin
Background: Separation of the pulmonic and systemic circulation is essential for terrestrial life, and mammals have evolved distinct cardiac chambers with specialized structures and functions. Transcriptomics profiling revealed cellular heterogeneity between heart chambers. However, the mechanisms underlying chamber-specific transcriptomic and metabolic differences-and their functional significance-remain poorly understood. The Hippo/YAP (yes-associated protein) pathway is a conserved signaling network that regulates diverse cellular processes. The Hippo kinases inhibit YAP in cardiac fibroblasts (CF) to restrict fibrosis and inflammation. Nonetheless, how YAP regulates the metabolic microenvironment during homeostasis and fibroinflammation remains unclear.
Methods: We investigated YAP and glycolysis activity in the 4 cardiac chambers by scoring the expression of YAP target genes and glycolysis genes in human single-nucleus RNA sequencing data. To compare glucose uptake between the left and right atria, we measured isotope-labeled glucose uptake in isolated mouse atria. To study the role of YAP in CFs, we inactivated the Hippo kinases, Lats1 and Lats2, in mouse CFs and performed metabolic studies, snRNA-seq, single-nucleus assay for transposase-accessible chromatin with sequencing, and spatial transcriptomics.
Results: Metabolic and sequencing approaches revealed that Hippo-deficient CFs activated glycolysis to promote fibroinflammation. Inhibition of glycolysis or lactate production suppressed Hippo-deficient CF-induced fibrosis. Elevated YAP activity disrupted fibroblast lineage fidelity by inducing an osteochondroprogenitor cell state. Blocking macrophage expansion pharmacologically reduced Hippo-deficient CF proliferation and fibrosis. Sequencing and functional studies showed that macrophages secreted IGF1 (insulin-like growth factor 1) to activate IGF1 signaling in Hippo-deficient CFs to increase cell proliferation and fibrosis.
Conclusions: We discovered that right atrial CFs are more glycolytic and have higher YAP activity than CFs in other heart chambers. YAP activation in CFs induces glycolysis to drive fibrosis. YAP disrupts fibroblast lineage fidelity, driving them to a SOX9 (SRY-box transcription factor 9)-expressing osteochondroprogenitor cell state. Mechanistically, YAP activates the secretion of CSF1 (colony-stimulating factor 1) to promote macrophage expansion. Blocking macrophage expansion reduces Hippo-deficient CF proliferation, osteochondroprogenitor differentiation, and fibrosis, revealing that macrophages signal reciprocally to regulate CF cell states. Genomic and functional studies revealed that the upregulated IGF1 receptor in Hippo-deficient CFs enables them to receive macrophage-secreted IGF1, thereby further enhancing CF proliferation and fibrosis.
{"title":"YAP-Induced Glycolysis Drives Fibroinflammation and Disrupts Fibroblast Fidelity.","authors":"Chang-Ru Tsai, Lin Liu, Yi Zhao, Jong H Kim, Paulo Czarnewski, Rich Gang Li, Fansen Meng, Mingjie Zheng, Jeffrey Steimle, Xiaolei Zhao, Francisco Grisanti, Zheng Sun, Jun Wang, Md Abul Hassan Samee, Xiao Li, James F Martin","doi":"10.1161/CIRCRESAHA.125.326480","DOIUrl":"10.1161/CIRCRESAHA.125.326480","url":null,"abstract":"<p><strong>Background: </strong>Separation of the pulmonic and systemic circulation is essential for terrestrial life, and mammals have evolved distinct cardiac chambers with specialized structures and functions. Transcriptomics profiling revealed cellular heterogeneity between heart chambers. However, the mechanisms underlying chamber-specific transcriptomic and metabolic differences-and their functional significance-remain poorly understood. The Hippo/YAP (yes-associated protein) pathway is a conserved signaling network that regulates diverse cellular processes. The Hippo kinases inhibit YAP in cardiac fibroblasts (CF) to restrict fibrosis and inflammation. Nonetheless, how YAP regulates the metabolic microenvironment during homeostasis and fibroinflammation remains unclear.</p><p><strong>Methods: </strong>We investigated YAP and glycolysis activity in the 4 cardiac chambers by scoring the expression of YAP target genes and glycolysis genes in human single-nucleus RNA sequencing data. To compare glucose uptake between the left and right atria, we measured isotope-labeled glucose uptake in isolated mouse atria. To study the role of YAP in CFs, we inactivated the Hippo kinases, <i>Lats1</i> and <i>Lats2</i>, in mouse CFs and performed metabolic studies, snRNA-seq, single-nucleus assay for transposase-accessible chromatin with sequencing, and spatial transcriptomics.</p><p><strong>Results: </strong>Metabolic and sequencing approaches revealed that Hippo-deficient CFs activated glycolysis to promote fibroinflammation. Inhibition of glycolysis or lactate production suppressed Hippo-deficient CF-induced fibrosis. Elevated YAP activity disrupted fibroblast lineage fidelity by inducing an osteochondroprogenitor cell state. Blocking macrophage expansion pharmacologically reduced Hippo-deficient CF proliferation and fibrosis. Sequencing and functional studies showed that macrophages secreted IGF1 (insulin-like growth factor 1) to activate IGF1 signaling in Hippo-deficient CFs to increase cell proliferation and fibrosis.</p><p><strong>Conclusions: </strong>We discovered that right atrial CFs are more glycolytic and have higher YAP activity than CFs in other heart chambers. YAP activation in CFs induces glycolysis to drive fibrosis. YAP disrupts fibroblast lineage fidelity, driving them to a SOX9 (SRY-box transcription factor 9)-expressing osteochondroprogenitor cell state. Mechanistically, YAP activates the secretion of CSF1 (colony-stimulating factor 1) to promote macrophage expansion. Blocking macrophage expansion reduces Hippo-deficient CF proliferation, osteochondroprogenitor differentiation, and fibrosis, revealing that macrophages signal reciprocally to regulate CF cell states. Genomic and functional studies revealed that the upregulated IGF1 receptor in Hippo-deficient CFs enables them to receive macrophage-secreted IGF1, thereby further enhancing CF proliferation and fibrosis.</p>","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":" ","pages":"1443-1458"},"PeriodicalIF":16.2,"publicationDate":"2025-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12965804/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145400088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-05Epub Date: 2025-11-06DOI: 10.1161/CIRCRESAHA.125.326391
Wenting Zhu, Ning Xie, Zhenyan Li, Xun Wang, Kuo Bi, Kun Zhu, Rilei Dai, Le Gao, Yufei Wang, Yang Li, Jing Guo, Lixuan Huang, Jingchen Li, Yingjiao Ju, Mingyang Li, Bing Hua, Weiwei An, Yangli Liu, Zhiheng Lin, Qinghua Cui, Chun-Mei Cao
Background: Peripheral artery disease is a severe ischemic vascular pathology without effective pharmacological approaches and improving angiogenesis to recover blood perfusion is a promising therapeutic strategy. Endothelial cells are the primary cell type contributing to angiogenesis in response to ischemia. However, the molecular mechanisms regulating ischemia-induced angiogenesis remain elusive.
Methods: We used a discovery-driven approach to identify elevated SRSF1 (serine/arginine splicing factor 1) expression in endothelial cells after ischemia. We used loss- and gain-of-function approaches to explore the role of SRSF1 in angiogenesis both in vivo and in vitro. A mouse model of hindlimb ischemia was used to evaluate ischemia-induced angiogenesis. We also investigated the mechanisms through transcriptome, enhanced crosslinking and immunoprecipitation sequencing, RNA pull-down, and chromatin immunoprecipitation-quantitative polymerase chain reaction analysis.
Results: Proteomic analyses identified endogenous SRSF1 accumulated in endothelial cells of the ischemic muscle and responded to hypoxia. Mice deficient in endothelial SRSF1 exhibited impaired blood flow recovery and impaired vasculature formation after hindlimb ischemia. Importantly, overexpression of SRSF1 enhanced blood flow recovery and angiogenesis after hindlimb ischemia. SRSF1 overexpression enhanced the angiogenic functions of human endothelial cells, promoting tube formation, sprouting capability, and cell migration, while SRSF1 knockdown suppressed these functions. Mechanistically, SRSF1 modulated the alternative splicing of ATF3 (activating transcription factor 3) by directly binding to ATF3 pre-mRNA (precursor messenger RNA), and SRSF1 overexpression elevated full-length ATF3 transcript at the expense of truncated ATF3Δzip2 transcript. ATF3 then bound directly to the KLF2 (Krüppel-like factor 2) promoter, suppressed KLF2 expression and downstream S1PR1 (sphingosine-1-phosphate receptor 1) signaling. Through upregulation of full-length ATF3 and downregulating KLF2-S1PR1 signaling, SRSF1 promoted endothelial tube formation and angiogenesis. In addition, alprostadil, the prostaglandin E1 analog, could activate the SRSF1 signaling to improve endothelial angiogenesis in vitro and in vivo.
Conclusions: Our findings identified SRSF1 as a novel regulator of ischemia-induced angiogenesis that enhances endothelial angiogenic functions by regulating the ATF3-KLF2-S1PR1 pathway. These results suggest that modulation of endothelial SRSF1 may represent a promising therapeutic approach for treating ischemic vascular diseases.
{"title":"Endothelial SRSF1 Promotes Ischemia-Induced Angiogenesis via ATF3-KLF2-S1PR1 Pathway.","authors":"Wenting Zhu, Ning Xie, Zhenyan Li, Xun Wang, Kuo Bi, Kun Zhu, Rilei Dai, Le Gao, Yufei Wang, Yang Li, Jing Guo, Lixuan Huang, Jingchen Li, Yingjiao Ju, Mingyang Li, Bing Hua, Weiwei An, Yangli Liu, Zhiheng Lin, Qinghua Cui, Chun-Mei Cao","doi":"10.1161/CIRCRESAHA.125.326391","DOIUrl":"10.1161/CIRCRESAHA.125.326391","url":null,"abstract":"<p><strong>Background: </strong>Peripheral artery disease is a severe ischemic vascular pathology without effective pharmacological approaches and improving angiogenesis to recover blood perfusion is a promising therapeutic strategy. Endothelial cells are the primary cell type contributing to angiogenesis in response to ischemia. However, the molecular mechanisms regulating ischemia-induced angiogenesis remain elusive.</p><p><strong>Methods: </strong>We used a discovery-driven approach to identify elevated SRSF1 (serine/arginine splicing factor 1) expression in endothelial cells after ischemia. We used loss- and gain-of-function approaches to explore the role of SRSF1 in angiogenesis both in vivo and in vitro. A mouse model of hindlimb ischemia was used to evaluate ischemia-induced angiogenesis. We also investigated the mechanisms through transcriptome, enhanced crosslinking and immunoprecipitation sequencing, RNA pull-down, and chromatin immunoprecipitation-quantitative polymerase chain reaction analysis.</p><p><strong>Results: </strong>Proteomic analyses identified endogenous SRSF1 accumulated in endothelial cells of the ischemic muscle and responded to hypoxia. Mice deficient in endothelial SRSF1 exhibited impaired blood flow recovery and impaired vasculature formation after hindlimb ischemia. Importantly, overexpression of SRSF1 enhanced blood flow recovery and angiogenesis after hindlimb ischemia. SRSF1 overexpression enhanced the angiogenic functions of human endothelial cells, promoting tube formation, sprouting capability, and cell migration, while SRSF1 knockdown suppressed these functions. Mechanistically, SRSF1 modulated the alternative splicing of ATF3 (activating transcription factor 3) by directly binding to ATF3 pre-mRNA (precursor messenger RNA), and SRSF1 overexpression elevated full-length ATF3 transcript at the expense of truncated ATF3Δzip2 transcript. ATF3 then bound directly to the KLF2 (Krüppel-like factor 2) promoter, suppressed KLF2 expression and downstream S1PR1 (sphingosine-1-phosphate receptor 1) signaling. Through upregulation of full-length ATF3 and downregulating KLF2-S1PR1 signaling, SRSF1 promoted endothelial tube formation and angiogenesis. In addition, alprostadil, the prostaglandin E1 analog, could activate the SRSF1 signaling to improve endothelial angiogenesis in vitro and in vivo.</p><p><strong>Conclusions: </strong>Our findings identified SRSF1 as a novel regulator of ischemia-induced angiogenesis that enhances endothelial angiogenic functions by regulating the ATF3-KLF2-S1PR1 pathway. These results suggest that modulation of endothelial SRSF1 may represent a promising therapeutic approach for treating ischemic vascular diseases.</p>","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":" ","pages":"1498-1521"},"PeriodicalIF":16.2,"publicationDate":"2025-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145451099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-04DOI: 10.1161/circresaha.125.326277
Chen-Shan Chen Woodcock,Giovanni Maroli,Hyunbum Kim,Yi Yin Tai,Ying Tang,Satoshi Okawa,Rami Homsi,Yunhye Kim,Shu-Ting Cho,Siyi Jiang,Caroline Chauvet,Bing Wang,Yassmin Al Aaraj,Robert Lafyatis,Rajan Saggar,John Sembrat,Qingde Wang,Qin Li,Andrea L Frump,Tim Lahm,Alexandra L McCubbrey,Tatiana V Kudryashova,Elena A Goncharova,Seyed Mehdi Nouraie,Thomas Bertero,Ke Yuan,Soni S Pullamsetti,Stephen Y Chan
BACKGROUNDEarly apoptosis of pulmonary artery endothelial cells (PAECs) is a driver of vascular remodeling and pulmonary hypertension (PH), but its regulation is poorly defined. ADAR1 (adenosine deaminase acting on RNA 1, gene name ADAR) is an RNA editing enzyme that converts adenosine to inosine in RNA transcripts and participates in RNA metabolism. Although deficiency in ADAR1-mediated RNA editing stimulates cellular innate immunity signaling and can promote apoptosis, the exact ADAR1 RNA editing targets and downstream mechanisms regulating PAEC survival are unknown. We sought to define the functions and targets of ADAR1-dependent RNA editing that control pulmonary endothelial pathophenotypes in PH.METHODS AND RESULTSADAR1 expression was downregulated in the pulmonary vascular endothelium and in the lung tissue of human and mouse PH. Global adenosine to inosine RNA editing was decreased in lungs from patients with PAH and hypoxic PH mice. In vitro, hypoxia, a PH trigger, downregulated ADAR1 in PAECs. By RNA sequencing of PAECs after ADAR1 knockdown, we identified the circadian gene NOCT (nocturnin) as a direct ADAR1 target. NOCT was found to carry 2 active adenosine-to-inosine RNA editing sites in the 3'UTR. By single-cell RNA sequencing of human PAH lungs, NOCT editing levels were reduced, while NOCT protein and transcript levels increased. Correspondingly, in vitro, ADAR1 silencing increased NOCT mRNA levels, thus inducing double-strand RNA-MDA5 (melanoma differentiation-associated protein 5) sensing interferon signaling and PAEC apoptosis. Importantly, silencing of NOCT reversed these changes. Forced NOCT expression phenocopied the effect of ADAR1 knockdown, upregulating interferon signaling molecules and increasing apoptosis. This ADAR1-NOCT axis was studied across multiple rodent models of disease. Chronically, hypoxic PH mice carrying a human missense mutation in ADAR displayed worsened PH. Forced adeno-associated virus expression of Adar improved monocrotaline-induced PH in rats. Genetic deletion of Noct mitigated PH in hypoxic interleukin 6-expressing transgenic PH mice, emphasizing the crucial role of NOCT in PH pathogenesis.CONCLUSIONSHypoxia-induced ADAR1 deficiency upregulates NOCT expression to induce PAEC interferon signaling activation, PAEC apoptosis, and PH. This study provides impetus to target the ADAR1-NOCT axis for more effective diagnostics and therapeutics for PH.
背景肺动脉内皮细胞（PAECs）的凋亡是血管重构和肺动脉高压（PH）的驱动因素，但其调控机制尚不明确。ADAR1 （adenosine deaminase acting on RNA 1，基因名ADAR）是一种RNA编辑酶，将RNA转录物中的腺苷转化为肌苷，参与RNA代谢。虽然缺乏ADAR1介导的RNA编辑可以刺激细胞先天免疫信号传导并促进细胞凋亡，但ADAR1 RNA编辑的确切靶点和调节PAEC存活的下游机制尚不清楚。我们试图确定adar1依赖性RNA编辑在PH中控制肺内皮病理表型的功能和靶点。方法和结果adar1在肺血管内皮和人和小鼠肺组织中的表达下调。PAH患者和缺氧PH小鼠肺中腺苷到肌苷RNA编辑减少。体外，缺氧，PH触发，下调paec中的ADAR1。通过对ADAR1敲低后paec的RNA测序，我们确定了昼夜节律基因NOCT （nocturnin）是ADAR1的直接靶点。发现NOCT在3'UTR中携带2个活跃的腺苷-肌苷RNA编辑位点。通过对人PAH肺的单细胞RNA测序，NOCT编辑水平降低，而NOCT蛋白和转录物水平升高。相应地，在体外，ADAR1沉默增加NOCT mRNA水平，从而诱导双链RNA-MDA5（黑色素瘤分化相关蛋白5）感知干扰素信号和PAEC凋亡。重要的是，NOCT的沉默逆转了这些变化。强制NOCT表达反映了ADAR1下调、干扰素信号分子上调和细胞凋亡增加的作用。在多种啮齿动物疾病模型中研究了ADAR1-NOCT轴。长期来看，携带人类ADAR错义突变的低氧PH小鼠的PH值恶化。强迫腺相关病毒表达ADAR可改善单鬼碱诱导的大鼠PH值。Noct基因缺失减轻了缺氧表达白细胞介素6转基因PH小鼠的PH，强调了Noct在PH发病中的重要作用。结论缺氧诱导的ADAR1缺失可上调NOCT表达，诱导PAEC干扰素信号激活、PAEC凋亡和PH。本研究为ADAR1-NOCT轴靶向治疗PH提供了更有效的诊断和治疗方法。
{"title":"Endothelial ADAR1 Deficit Induces the NOCT-IRF7 Axis in Pulmonary Hypertension.","authors":"Chen-Shan Chen Woodcock,Giovanni Maroli,Hyunbum Kim,Yi Yin Tai,Ying Tang,Satoshi Okawa,Rami Homsi,Yunhye Kim,Shu-Ting Cho,Siyi Jiang,Caroline Chauvet,Bing Wang,Yassmin Al Aaraj,Robert Lafyatis,Rajan Saggar,John Sembrat,Qingde Wang,Qin Li,Andrea L Frump,Tim Lahm,Alexandra L McCubbrey,Tatiana V Kudryashova,Elena A Goncharova,Seyed Mehdi Nouraie,Thomas Bertero,Ke Yuan,Soni S Pullamsetti,Stephen Y Chan","doi":"10.1161/circresaha.125.326277","DOIUrl":"https://doi.org/10.1161/circresaha.125.326277","url":null,"abstract":"BACKGROUNDEarly apoptosis of pulmonary artery endothelial cells (PAECs) is a driver of vascular remodeling and pulmonary hypertension (PH), but its regulation is poorly defined. ADAR1 (adenosine deaminase acting on RNA 1, gene name ADAR) is an RNA editing enzyme that converts adenosine to inosine in RNA transcripts and participates in RNA metabolism. Although deficiency in ADAR1-mediated RNA editing stimulates cellular innate immunity signaling and can promote apoptosis, the exact ADAR1 RNA editing targets and downstream mechanisms regulating PAEC survival are unknown. We sought to define the functions and targets of ADAR1-dependent RNA editing that control pulmonary endothelial pathophenotypes in PH.METHODS AND RESULTSADAR1 expression was downregulated in the pulmonary vascular endothelium and in the lung tissue of human and mouse PH. Global adenosine to inosine RNA editing was decreased in lungs from patients with PAH and hypoxic PH mice. In vitro, hypoxia, a PH trigger, downregulated ADAR1 in PAECs. By RNA sequencing of PAECs after ADAR1 knockdown, we identified the circadian gene NOCT (nocturnin) as a direct ADAR1 target. NOCT was found to carry 2 active adenosine-to-inosine RNA editing sites in the 3'UTR. By single-cell RNA sequencing of human PAH lungs, NOCT editing levels were reduced, while NOCT protein and transcript levels increased. Correspondingly, in vitro, ADAR1 silencing increased NOCT mRNA levels, thus inducing double-strand RNA-MDA5 (melanoma differentiation-associated protein 5) sensing interferon signaling and PAEC apoptosis. Importantly, silencing of NOCT reversed these changes. Forced NOCT expression phenocopied the effect of ADAR1 knockdown, upregulating interferon signaling molecules and increasing apoptosis. This ADAR1-NOCT axis was studied across multiple rodent models of disease. Chronically, hypoxic PH mice carrying a human missense mutation in ADAR displayed worsened PH. Forced adeno-associated virus expression of Adar improved monocrotaline-induced PH in rats. Genetic deletion of Noct mitigated PH in hypoxic interleukin 6-expressing transgenic PH mice, emphasizing the crucial role of NOCT in PH pathogenesis.CONCLUSIONSHypoxia-induced ADAR1 deficiency upregulates NOCT expression to induce PAEC interferon signaling activation, PAEC apoptosis, and PH. This study provides impetus to target the ADAR1-NOCT axis for more effective diagnostics and therapeutics for PH.","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":"1 1","pages":""},"PeriodicalIF":20.1,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145664178","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-04DOI: 10.1161/circresaha.125.327610
Luis A Gonano,Cecilia Mundiña-Weilenmann
{"title":"Ca²⁺ in the Mitochondrial Intermembrane Space: A New Compartment Fueling Arrhythmias.","authors":"Luis A Gonano,Cecilia Mundiña-Weilenmann","doi":"10.1161/circresaha.125.327610","DOIUrl":"https://doi.org/10.1161/circresaha.125.327610","url":null,"abstract":"","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":"26 1","pages":"1404-1406"},"PeriodicalIF":20.1,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145673957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-04DOI: 10.1161/circresaha.125.327486
Robert D Hume,Jessica Warwick,Woo Jun Shim,Cassandra Malecki,Mengbo Li,Lakshay Seth,Dylan Harney,Julien Dagher,Trina Lum,Geraldine Tierney,Wendy Cooper,Eugene Slaughter,Xiaosuo Wang,Lisa Nguyen,Louise Cole,James Edelman,Fairooj Rashid,Callum Houlahan,Antony Gao,Angela L Ferguson,James J H Chong,Mark Larance,John O'Sullivan,Nathan J Palpant,Paul Bannon,Sean Lal
BACKGROUNDMyocardial infarction (MI) is a leading cause of death worldwide and can eliminate up to a third of the cardiomyocytes within the human heart. Although cardiomyocytes undergo mitosis during early development, most cardiomyocytes cease cell cycling soon after birth. In contrast, rodent MI models have shown that cardiomyocytes increase mitosis in response to ischemia; however, this has not been shown in humans.METHODSUsing a unique premortem post-MI human heart, immunostaining, bulk RNA sequencing, proteomics, metabolomics, single-nucleus RNA sequencing and a novel post-MI human biopsy method, we investigated human cardiomyocyte mitosis post-MI.RESULTSWe show that adult human cardiomyocytes exhibit increased mitosis and cytokinesis in response to ischemia.CONCLUSIONSFuture development of therapeutics to enhance this intrinsic mitotic potential could lead to new treatments that reverse heart failure via cardiac regeneration.
{"title":"Human Hearts Intrinsically Increase Cardiomyocyte Mitosis After Myocardial Infarction.","authors":"Robert D Hume,Jessica Warwick,Woo Jun Shim,Cassandra Malecki,Mengbo Li,Lakshay Seth,Dylan Harney,Julien Dagher,Trina Lum,Geraldine Tierney,Wendy Cooper,Eugene Slaughter,Xiaosuo Wang,Lisa Nguyen,Louise Cole,James Edelman,Fairooj Rashid,Callum Houlahan,Antony Gao,Angela L Ferguson,James J H Chong,Mark Larance,John O'Sullivan,Nathan J Palpant,Paul Bannon,Sean Lal","doi":"10.1161/circresaha.125.327486","DOIUrl":"https://doi.org/10.1161/circresaha.125.327486","url":null,"abstract":"BACKGROUNDMyocardial infarction (MI) is a leading cause of death worldwide and can eliminate up to a third of the cardiomyocytes within the human heart. Although cardiomyocytes undergo mitosis during early development, most cardiomyocytes cease cell cycling soon after birth. In contrast, rodent MI models have shown that cardiomyocytes increase mitosis in response to ischemia; however, this has not been shown in humans.METHODSUsing a unique premortem post-MI human heart, immunostaining, bulk RNA sequencing, proteomics, metabolomics, single-nucleus RNA sequencing and a novel post-MI human biopsy method, we investigated human cardiomyocyte mitosis post-MI.RESULTSWe show that adult human cardiomyocytes exhibit increased mitosis and cytokinesis in response to ischemia.CONCLUSIONSFuture development of therapeutics to enhance this intrinsic mitotic potential could lead to new treatments that reverse heart failure via cardiac regeneration.","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":"198200 1","pages":""},"PeriodicalIF":20.1,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145664181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-04DOI: 10.1161/circresaha.125.327497
Eleonora Torre,Mélanie Faure,Isabelle Bidaud,Matthias Baudot,Marvin Gaillardon,Walma Pereira de Vasconcelos,Sihame Laarioui,Leïla Talssi,Birgit Engeland,Steven Reiken,Andrea Saponaro,Bi-Xing Chen,Anna Moroni,Alicia D'Souza,Dirk Isbrandt,Andrew R Marks,Steven O Marx,Pietro Mesirca,Matteo E Mangoni
BACKGROUNDThe ionic mechanism by which catecholamines increase the heart rate is incompletely understood. In this study, we have assessed the roles of sinoatrial node L-type Cav1.3 (α1D) Ca2+ channels, phosphorylation of L-type channel regulatory partner protein Rad (Ras-related RGK G-protein), and cAMP-dependent regulation of hyperpolarization-activated HCN (hyperpolarization-activated) channels.METHODSWe studied β-adrenergic regulation of heart rate and sinoatrial pacemaker activity in mice lacking Cav1.3 channels and in mice expressing dihydropyridine-insensitive L-type Cav1.2 channels alone or concomitantly expressing cAMP-insensitive HCN4 subunits in a heart-specific and time-controlled manner. We also studied the chronotropic response to sympathomimetics of sinoatrial pacemaker myocytes under conditions of specific inhibition of cAMP-dependent regulation of HCN4 by the cyclic dinucleotide cyclic di-(3',5')-GMP and ablation of PKA (protein kinase A)-dependent phosphorylation of Rad.RESULTSMutant mice with knockout of Cav1.3 and cAMP-insensitive HCN4 subunits in the heart lacked diurnal variation in heart rate and failed to increase their heart rate after administration of catecholamines or during physical activity. Selective pharmacological inhibition of Cav1.3 prevented the enhancement of pacemaker activity by sympathomimetics or by direct activation of adenylate cyclase, as well as by phosphodiesterase inhibitors, when cAMP-dependent regulation of HCN was simultaneously silenced. Upregulation of Cav1.3 and HCN-mediated funny current (If) accounted for the total change in diastolic current on activation of β-adrenoceptors, explaining the loss of chronotropic effect of catecholamines. Preventing PKA phosphorylation of Rad abrogated the chronotropic response to sympathomimetics of intact hearts and of pacemaker myocytes under HCN blockade, or cAMP-dependent regulation of HCN4, respectively.CONCLUSIONSPKA phosphorylation of Rad, which disinhibits Cav1.3 channels and cAMP-dependent activation of HCN channels, are key effectors in β-adrenergic regulation of cardiac pacemaker activity and can sustain positive chronotropic effects independently. These findings on Rad-mediated regulation of Cav1.3 and HCN channels unravel the ionic mechanisms underlying the catecholaminergic acceleration of the heart rate.
{"title":"L-Type Cav1.3 and HCN Channels Mediate Heart Rate Acceleration by Catecholamines.","authors":"Eleonora Torre,Mélanie Faure,Isabelle Bidaud,Matthias Baudot,Marvin Gaillardon,Walma Pereira de Vasconcelos,Sihame Laarioui,Leïla Talssi,Birgit Engeland,Steven Reiken,Andrea Saponaro,Bi-Xing Chen,Anna Moroni,Alicia D'Souza,Dirk Isbrandt,Andrew R Marks,Steven O Marx,Pietro Mesirca,Matteo E Mangoni","doi":"10.1161/circresaha.125.327497","DOIUrl":"https://doi.org/10.1161/circresaha.125.327497","url":null,"abstract":"BACKGROUNDThe ionic mechanism by which catecholamines increase the heart rate is incompletely understood. In this study, we have assessed the roles of sinoatrial node L-type Cav1.3 (α1D) Ca2+ channels, phosphorylation of L-type channel regulatory partner protein Rad (Ras-related RGK G-protein), and cAMP-dependent regulation of hyperpolarization-activated HCN (hyperpolarization-activated) channels.METHODSWe studied β-adrenergic regulation of heart rate and sinoatrial pacemaker activity in mice lacking Cav1.3 channels and in mice expressing dihydropyridine-insensitive L-type Cav1.2 channels alone or concomitantly expressing cAMP-insensitive HCN4 subunits in a heart-specific and time-controlled manner. We also studied the chronotropic response to sympathomimetics of sinoatrial pacemaker myocytes under conditions of specific inhibition of cAMP-dependent regulation of HCN4 by the cyclic dinucleotide cyclic di-(3',5')-GMP and ablation of PKA (protein kinase A)-dependent phosphorylation of Rad.RESULTSMutant mice with knockout of Cav1.3 and cAMP-insensitive HCN4 subunits in the heart lacked diurnal variation in heart rate and failed to increase their heart rate after administration of catecholamines or during physical activity. Selective pharmacological inhibition of Cav1.3 prevented the enhancement of pacemaker activity by sympathomimetics or by direct activation of adenylate cyclase, as well as by phosphodiesterase inhibitors, when cAMP-dependent regulation of HCN was simultaneously silenced. Upregulation of Cav1.3 and HCN-mediated funny current (If) accounted for the total change in diastolic current on activation of β-adrenoceptors, explaining the loss of chronotropic effect of catecholamines. Preventing PKA phosphorylation of Rad abrogated the chronotropic response to sympathomimetics of intact hearts and of pacemaker myocytes under HCN blockade, or cAMP-dependent regulation of HCN4, respectively.CONCLUSIONSPKA phosphorylation of Rad, which disinhibits Cav1.3 channels and cAMP-dependent activation of HCN channels, are key effectors in β-adrenergic regulation of cardiac pacemaker activity and can sustain positive chronotropic effects independently. These findings on Rad-mediated regulation of Cav1.3 and HCN channels unravel the ionic mechanisms underlying the catecholaminergic acceleration of the heart rate.","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":"7 1","pages":""},"PeriodicalIF":20.1,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145664180","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BACKGROUNDStroke remains a leading cause of mortality and disability, driven by complex, time-dependent mechanisms that aggravate ischemic injury. Collateral perfusion dictates infarct size, expansion rate, and penumbral preservation, yet its regulation is poorly understood. Beyond structural/genetic factors such as aging or cardiovascular risk, functional influences like circadian immune activity may also affect vascular patency. Neutrophils, key mediators of ischemic injury, exhibit circadian oscillations in phenotype and function that could modulate collateral flow and stroke outcome.METHODSWe combined permanent and transient middle cerebral artery occlusion models in mice with flow cytometry, single-cell RNA sequencing, confocal microscopy, and laser speckle imaging to investigate time-of-day-dependent neutrophil mechanisms in stroke. Pharmacological (chloramidine, DNase-I) and genetic (Pad4 [peptidyl arginine deiminase 4]-/-, Bmal1 [brain and muscle ARNT-like 1]Neu, Cxcr4 [C-X-C chemokine receptor type 4]Neu) interventions were used to define how time-of-day regulation shapes neutrophil function, net extracellular traps (NETs) formation, and stroke severity. A cohort of 540 patients with ischemic stroke was analyzed for diurnal patterns of NET-related biomarkers and their association with collateral circulation and clinical outcomes.RESULTSInfarct volume and neurological deficits exhibited clear circadian oscillations, with worse outcomes when stroke occurred during the murine inactive phase (Zeitgeber time 5) versus the active phase (Zeitgeber time 13). These fluctuations disappeared after neutrophil depletion or clock disruption. During the inactive phase, neutrophils displayed an activated, NET-prone phenotype, causing microvascular staling and reduced collateral perfusion. Inhibiting NET formation pharmacologically or through Pad4 deletion restored perfusion and abolished time-of-day effects. In patients, neutrophil and NET-related biomarkers (MPO [myeloperoxidase], elastase, sCD40L [soluble CD40 ligand]) showed diurnal oscillations, peaking during the human inactive phase (evening/night), coinciding with reduced collateral flow and poorer outcomes.CONCLUSIONSTime-of-day regulation of neutrophil function critically determines collateral perfusion and stroke severity. Neutrophil-driven NETosis during the inactive phase promotes microvascular obstruction and worsens outcomes. Targeting NET formation or timing therapy could enhance collateral efficacy and offer novel chronotherapeutic opportunities for stroke treatment.
{"title":"Circadian Control of Neutrophils Drives Collateral Perfusion and Stroke Outcome.","authors":"Sandra Vázquez-Reyes,Alicia García-Culebras,Gaohong Di,Francisco J De Castro-Millán,Blanca Díaz-Benito,Carmen Nieto-Vaquero,Alessandra Ruiz-Sanchez,Eneko Merino-Casamayor,Carlos Parra-Pérez,Ana Moraga,César Core-Barrera,Patricia Calleja,Ana Dopazo,Sergio Callejas,Andrea Rubio-Ponce,Alejandra Aroca-Crevillén,Fátima Sánchez-Cabo,Sara Pascual El Bobakry,Carlos Torroja,Elga Esposito,Eng H Lo,Iván Ballesteros,Andrés Hidalgo,María Isabel Cuartero,Ignacio Lizasoain,María Ángeles Moro","doi":"10.1161/circresaha.125.326438","DOIUrl":"https://doi.org/10.1161/circresaha.125.326438","url":null,"abstract":"BACKGROUNDStroke remains a leading cause of mortality and disability, driven by complex, time-dependent mechanisms that aggravate ischemic injury. Collateral perfusion dictates infarct size, expansion rate, and penumbral preservation, yet its regulation is poorly understood. Beyond structural/genetic factors such as aging or cardiovascular risk, functional influences like circadian immune activity may also affect vascular patency. Neutrophils, key mediators of ischemic injury, exhibit circadian oscillations in phenotype and function that could modulate collateral flow and stroke outcome.METHODSWe combined permanent and transient middle cerebral artery occlusion models in mice with flow cytometry, single-cell RNA sequencing, confocal microscopy, and laser speckle imaging to investigate time-of-day-dependent neutrophil mechanisms in stroke. Pharmacological (chloramidine, DNase-I) and genetic (Pad4 [peptidyl arginine deiminase 4]-/-, Bmal1 [brain and muscle ARNT-like 1]Neu, Cxcr4 [C-X-C chemokine receptor type 4]Neu) interventions were used to define how time-of-day regulation shapes neutrophil function, net extracellular traps (NETs) formation, and stroke severity. A cohort of 540 patients with ischemic stroke was analyzed for diurnal patterns of NET-related biomarkers and their association with collateral circulation and clinical outcomes.RESULTSInfarct volume and neurological deficits exhibited clear circadian oscillations, with worse outcomes when stroke occurred during the murine inactive phase (Zeitgeber time 5) versus the active phase (Zeitgeber time 13). These fluctuations disappeared after neutrophil depletion or clock disruption. During the inactive phase, neutrophils displayed an activated, NET-prone phenotype, causing microvascular staling and reduced collateral perfusion. Inhibiting NET formation pharmacologically or through Pad4 deletion restored perfusion and abolished time-of-day effects. In patients, neutrophil and NET-related biomarkers (MPO [myeloperoxidase], elastase, sCD40L [soluble CD40 ligand]) showed diurnal oscillations, peaking during the human inactive phase (evening/night), coinciding with reduced collateral flow and poorer outcomes.CONCLUSIONSTime-of-day regulation of neutrophil function critically determines collateral perfusion and stroke severity. Neutrophil-driven NETosis during the inactive phase promotes microvascular obstruction and worsens outcomes. Targeting NET formation or timing therapy could enhance collateral efficacy and offer novel chronotherapeutic opportunities for stroke treatment.","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":"18 1","pages":""},"PeriodicalIF":20.1,"publicationDate":"2025-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145599790","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-26DOI: 10.1161/circresaha.125.326628
Maria Viskadourou,Sharjeel Chaudhry,Arianna Scalco,Paula Reventun,Pablo Toledano-Sanz,Roujin An,Nunzio Alcharani,Maria Delgado Marin,Abigail Fennell,Quanyi Zhao,William Osburn,Andrew S McCallion,Thomas Quertermous,Alexis Battle,Charles J Lowenstein,Marios Arvanitis
BACKGROUNDGenome-wide association studies have identified multiple novel loci that contribute to coronary artery disease pathogenesis, but the mechanisms of these associations remain largely unknown.METHODSIn this study, we used a multitrait colocalization approach to prioritize novel endothelial-specific loci for atherosclerosis. We combined computational methods with in vitro assays and mouse models to study one of those new loci targeting the gene REST.RESULTSA multitrait colocalization approach across expression quantitative trait loci in atherosclerosis-relevant cell types, followed by in vitro CRISPR interference, revealed that a conserved regulatory element in a chromosome 4 genetic locus increases the risk of coronary artery disease and decreases the expression of REST, a transcriptional repressor, in endothelial cells. Pcsk9-overexpressing mice with an endothelial-specific knockout of Rest exhibited increased atherosclerotic plaque formation in their aortas, with increased macrophage and lipid deposition within the plaque after 16 weeks of high-fat diet exposure compared with littermate controls. RNA-seq in human aortic endothelial cells after REST silencing, followed by assessment of protein expression, revealed that REST silencing triggers endothelial-to-mesenchymal transition. Consistently, REST silencing increased endothelial permeability and migration in vitro. Single-nucleus RNA sequencing in endothelial lineage traced atherosclerotic mice with Rest knock-out revealed evidence of endothelial TGFb signaling activation and of transition smooth muscle-like cells in atherosclerotic aortas on genetic knockout of Rest. cleavage under targets and tagmentation (CUT&Tag) sequencing did not identify any known TGFb effector genes as direct REST transcriptional targets. Instead, joint analysis of CUT&Tag with RNA-seq highlighted L1CAM, a known endothelial-to-mesenchymal transition activator, and its interactors as the most significant gene set directly affected by REST in the endothelium. Simultaneous silencing of L1CAM and REST in human aortic endothelial cells inhibited the upregulation of mesenchymal genes and the enhanced migration induced by REST silencing and diminished the upregulation of several TGFb effectors overexpressed on REST silencing.CONCLUSIONSIn summary, our data reveal the novel role of REST as a repressor in endothelial cells that functions to constitutively inhibit endothelial-to-mesenchymal transition and protect against atherosclerosis.
{"title":"Functional Genomics Link REST to Endothelial Plasticity and Atherosclerosis.","authors":"Maria Viskadourou,Sharjeel Chaudhry,Arianna Scalco,Paula Reventun,Pablo Toledano-Sanz,Roujin An,Nunzio Alcharani,Maria Delgado Marin,Abigail Fennell,Quanyi Zhao,William Osburn,Andrew S McCallion,Thomas Quertermous,Alexis Battle,Charles J Lowenstein,Marios Arvanitis","doi":"10.1161/circresaha.125.326628","DOIUrl":"https://doi.org/10.1161/circresaha.125.326628","url":null,"abstract":"BACKGROUNDGenome-wide association studies have identified multiple novel loci that contribute to coronary artery disease pathogenesis, but the mechanisms of these associations remain largely unknown.METHODSIn this study, we used a multitrait colocalization approach to prioritize novel endothelial-specific loci for atherosclerosis. We combined computational methods with in vitro assays and mouse models to study one of those new loci targeting the gene REST.RESULTSA multitrait colocalization approach across expression quantitative trait loci in atherosclerosis-relevant cell types, followed by in vitro CRISPR interference, revealed that a conserved regulatory element in a chromosome 4 genetic locus increases the risk of coronary artery disease and decreases the expression of REST, a transcriptional repressor, in endothelial cells. Pcsk9-overexpressing mice with an endothelial-specific knockout of Rest exhibited increased atherosclerotic plaque formation in their aortas, with increased macrophage and lipid deposition within the plaque after 16 weeks of high-fat diet exposure compared with littermate controls. RNA-seq in human aortic endothelial cells after REST silencing, followed by assessment of protein expression, revealed that REST silencing triggers endothelial-to-mesenchymal transition. Consistently, REST silencing increased endothelial permeability and migration in vitro. Single-nucleus RNA sequencing in endothelial lineage traced atherosclerotic mice with Rest knock-out revealed evidence of endothelial TGFb signaling activation and of transition smooth muscle-like cells in atherosclerotic aortas on genetic knockout of Rest. cleavage under targets and tagmentation (CUT&Tag) sequencing did not identify any known TGFb effector genes as direct REST transcriptional targets. Instead, joint analysis of CUT&Tag with RNA-seq highlighted L1CAM, a known endothelial-to-mesenchymal transition activator, and its interactors as the most significant gene set directly affected by REST in the endothelium. Simultaneous silencing of L1CAM and REST in human aortic endothelial cells inhibited the upregulation of mesenchymal genes and the enhanced migration induced by REST silencing and diminished the upregulation of several TGFb effectors overexpressed on REST silencing.CONCLUSIONSIn summary, our data reveal the novel role of REST as a repressor in endothelial cells that functions to constitutively inhibit endothelial-to-mesenchymal transition and protect against atherosclerosis.","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":"17 1","pages":""},"PeriodicalIF":20.1,"publicationDate":"2025-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145599640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-21DOI: 10.1161/circresaha.125.326647
Lu Gao,Jinhua Cao,Yue Li,Xiaoyang Ji,Qingqing Wu,Sen Guo,Xintong Cai,Ke Li,Yanna Sun,Lili Xiao,Youyou Du,Zhe Zheng,Xiaofang Wang
BACKGROUNDCardiac hypertrophy is one of the major causes of heart failure and sudden cardiac death. OTUD7a (OTU domain-containing protein 7a) is identified as a deubiquitinizing enzyme and a possible tumor suppressor. The present study is aimed at exploring the potential role and key downstream effectors of OTUD7a in cardiac hypertrophy.METHODSThe expression level of OTUD7a was detected in the cardiomyocytes with phenylephrine stimuli and the hearts subjected to transverse aortic constriction surgery. Then, the potential effects of OTUD7a on cardiac hypertrophy were evaluated in vivo by using cardiac-specific OTUD7a knockout mice and adeno-associated virus serotype 9-OTUD7a-infected mice. To further explore the direct modulation of OTUD7a on cardiomyocytes, hypertrophic parameters were detected in phenylephrine-stimulated cardiomyocytes with adenovirus system-induced OTUD7a overexpression or depletion. Furthermore, RNA-sequencing and interactome analysis, which were followed by multiple molecular biological methodologies, were combined to identify the direct target and corresponding molecular events contributing to OTUD7a function.RESULTSCardiac hypertrophy stimulates expression of OTUD7a in vitro and in vivo. Our data clearly showed that OTUD7a deficiency alleviates pathological cardiac hypertrophy in the transverse aortic constriction mouse model as well as in phenylephrine-treated cardiomyocytes, whereas overexpression of OTUD7a aggravated hypertrophic heart in vivo and enhanced cardiomyocyte enlargement in vitro. Mechanistically, TAK1 (transforming growth factor-β-activated kinase 1) was identified as a direct and essential target of OTUD7a in cardiac hypertrophy. To be more specific, OTUD7a directly interacts with TAK1 to inhibit the ubiquitination degradation of TAK1 and subsequently increase the phosphorylation levels of TAK1 and its downstream JNK (c-Jun N-terminal kinase)/P38. 5Z-7-oxozeaenol, a TAK1 inhibitor, blocked the detrimental effects of OTUD7a. Moreover, overexpression of TAK1 abolished the protection of OTUD7a depletion.CONCLUSIONSOur findings, for the first time, provide evidence supporting OTUD7a as a novel promoter of pathological cardiac hypertrophy and indicate that targeting the OTUD7a-TAK1 axis represents a promising therapeutic strategy for cardiac hypertrophy and related heart failure.
{"title":"OTUD7a Accelerates Pathological Cardiac Hypertrophy via TAK1 Activation.","authors":"Lu Gao,Jinhua Cao,Yue Li,Xiaoyang Ji,Qingqing Wu,Sen Guo,Xintong Cai,Ke Li,Yanna Sun,Lili Xiao,Youyou Du,Zhe Zheng,Xiaofang Wang","doi":"10.1161/circresaha.125.326647","DOIUrl":"https://doi.org/10.1161/circresaha.125.326647","url":null,"abstract":"BACKGROUNDCardiac hypertrophy is one of the major causes of heart failure and sudden cardiac death. OTUD7a (OTU domain-containing protein 7a) is identified as a deubiquitinizing enzyme and a possible tumor suppressor. The present study is aimed at exploring the potential role and key downstream effectors of OTUD7a in cardiac hypertrophy.METHODSThe expression level of OTUD7a was detected in the cardiomyocytes with phenylephrine stimuli and the hearts subjected to transverse aortic constriction surgery. Then, the potential effects of OTUD7a on cardiac hypertrophy were evaluated in vivo by using cardiac-specific OTUD7a knockout mice and adeno-associated virus serotype 9-OTUD7a-infected mice. To further explore the direct modulation of OTUD7a on cardiomyocytes, hypertrophic parameters were detected in phenylephrine-stimulated cardiomyocytes with adenovirus system-induced OTUD7a overexpression or depletion. Furthermore, RNA-sequencing and interactome analysis, which were followed by multiple molecular biological methodologies, were combined to identify the direct target and corresponding molecular events contributing to OTUD7a function.RESULTSCardiac hypertrophy stimulates expression of OTUD7a in vitro and in vivo. Our data clearly showed that OTUD7a deficiency alleviates pathological cardiac hypertrophy in the transverse aortic constriction mouse model as well as in phenylephrine-treated cardiomyocytes, whereas overexpression of OTUD7a aggravated hypertrophic heart in vivo and enhanced cardiomyocyte enlargement in vitro. Mechanistically, TAK1 (transforming growth factor-β-activated kinase 1) was identified as a direct and essential target of OTUD7a in cardiac hypertrophy. To be more specific, OTUD7a directly interacts with TAK1 to inhibit the ubiquitination degradation of TAK1 and subsequently increase the phosphorylation levels of TAK1 and its downstream JNK (c-Jun N-terminal kinase)/P38. 5Z-7-oxozeaenol, a TAK1 inhibitor, blocked the detrimental effects of OTUD7a. Moreover, overexpression of TAK1 abolished the protection of OTUD7a depletion.CONCLUSIONSOur findings, for the first time, provide evidence supporting OTUD7a as a novel promoter of pathological cardiac hypertrophy and indicate that targeting the OTUD7a-TAK1 axis represents a promising therapeutic strategy for cardiac hypertrophy and related heart failure.","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":"105 1","pages":""},"PeriodicalIF":20.1,"publicationDate":"2025-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145559212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-19DOI: 10.1161/circresaha.125.326412
James B Hayes,Dharmendra Choudhary,Dylan Ritter,Abigail C Neininger-Castro,Alaina H Willet,Leah R Caplan,Yu Wang,Xiao Liu,Nilay Taneja,Zachary C Sanchez,Kyra Smart,David W J Armstrong,Cynthia A Reinhart-King,Qi Liu,Matthew J Tyska,Erdem D Tabdanov,W David Merryman,Quinn S Wells,Ela W Knapik,Dylan T Burnette
BACKGROUNDCardiac sarcomeres generate the fundamental forces of each heartbeat. Cardiac myocytes (CMs) express nonmuscle versions of muscle-specific sarcomere proteins, which have unknown relevance to sarcomere function or heart physiology.METHODSExpression levels of nonmuscle cytoskeletal proteins versus muscle-specific counterparts in CMs were directly compared. Function and subcellular localization of the nonmuscle protein ACTN4 (alpha-actinin 4) in induced pluripotent stem cell-derived CMs were determined using small interfering RNA-mediated knockdown, overexpression, and pharmacological perturbation. Impacts of ACTN4 depletion or knockout on cardiac structure function were evaluated in zebrafish embryos. Left ventricular actn4 levels were evaluated in a mouse model of chronic pressure overload. Human ACTN4 gene variants were tested for association with heart failure with preserved ejection fraction using the BioVU biobank. A meta-analysis was conducted on ventricular data sets of human cardiomyopathies.RESULTSACTN4 expression in human CMs met or exceeded some muscle-specific genes (eg, MYH6). Anti-ACTN4 antibodies colocalized with anti-ACTN2 (alpha-actinin 2) at the sarcomere Z-disc in human, mouse, and zebrafish ventricular tissue. Coimmunoprecipitation and structural modeling suggest a Z-disc ACTN4:ACTN2 complex. ACTN4 depletion from induced pluripotent stem cell-derived CMs resulted in increased sarcomere assembly, decreased sarcomere component turnover, elevated contractile force, and contractility-dependent cellular hypertrophy. Overexpression of an ACTN4 actin-binding chimera suppressed sarcomere assembly. In zebrafish embryos, ACTN4 depletion/knockout induced ventricular hypercontractility and atrial enlargement. Selective modulation of ventricular contractility was sufficient to prevent or phenocopy atrial remodeling. In mice, actn4, but not actn2, was upregulated in the left ventricular following pressure overload. One of 14 ACTN4 single-nucleotide polymorphisms was associated with reduced heart failure with preserved ejection fraction risk in humans, and prepublished studies suggest a pattern of ventricular ACTN4 upregulation in certain human cardiomyopathies.CONCLUSIONSA nonmuscle actinin (ACTN4) populates the cardiac Z-disc. ACTN4 regulates sarcomeric architecture in CMs. ACTN4 influences fractional shortening at the cell level and contractility at the tissue level. Changes in ventricular ACTN4 levels are associated with remodeling and may influence clinical outcomes related to heart failure.
{"title":"Non-Muscle α-Actinin-4 Couples Sarcomere Function to Cardiac Remodeling.","authors":"James B Hayes,Dharmendra Choudhary,Dylan Ritter,Abigail C Neininger-Castro,Alaina H Willet,Leah R Caplan,Yu Wang,Xiao Liu,Nilay Taneja,Zachary C Sanchez,Kyra Smart,David W J Armstrong,Cynthia A Reinhart-King,Qi Liu,Matthew J Tyska,Erdem D Tabdanov,W David Merryman,Quinn S Wells,Ela W Knapik,Dylan T Burnette","doi":"10.1161/circresaha.125.326412","DOIUrl":"https://doi.org/10.1161/circresaha.125.326412","url":null,"abstract":"BACKGROUNDCardiac sarcomeres generate the fundamental forces of each heartbeat. Cardiac myocytes (CMs) express nonmuscle versions of muscle-specific sarcomere proteins, which have unknown relevance to sarcomere function or heart physiology.METHODSExpression levels of nonmuscle cytoskeletal proteins versus muscle-specific counterparts in CMs were directly compared. Function and subcellular localization of the nonmuscle protein ACTN4 (alpha-actinin 4) in induced pluripotent stem cell-derived CMs were determined using small interfering RNA-mediated knockdown, overexpression, and pharmacological perturbation. Impacts of ACTN4 depletion or knockout on cardiac structure function were evaluated in zebrafish embryos. Left ventricular actn4 levels were evaluated in a mouse model of chronic pressure overload. Human ACTN4 gene variants were tested for association with heart failure with preserved ejection fraction using the BioVU biobank. A meta-analysis was conducted on ventricular data sets of human cardiomyopathies.RESULTSACTN4 expression in human CMs met or exceeded some muscle-specific genes (eg, MYH6). Anti-ACTN4 antibodies colocalized with anti-ACTN2 (alpha-actinin 2) at the sarcomere Z-disc in human, mouse, and zebrafish ventricular tissue. Coimmunoprecipitation and structural modeling suggest a Z-disc ACTN4:ACTN2 complex. ACTN4 depletion from induced pluripotent stem cell-derived CMs resulted in increased sarcomere assembly, decreased sarcomere component turnover, elevated contractile force, and contractility-dependent cellular hypertrophy. Overexpression of an ACTN4 actin-binding chimera suppressed sarcomere assembly. In zebrafish embryos, ACTN4 depletion/knockout induced ventricular hypercontractility and atrial enlargement. Selective modulation of ventricular contractility was sufficient to prevent or phenocopy atrial remodeling. In mice, actn4, but not actn2, was upregulated in the left ventricular following pressure overload. One of 14 ACTN4 single-nucleotide polymorphisms was associated with reduced heart failure with preserved ejection fraction risk in humans, and prepublished studies suggest a pattern of ventricular ACTN4 upregulation in certain human cardiomyopathies.CONCLUSIONSA nonmuscle actinin (ACTN4) populates the cardiac Z-disc. ACTN4 regulates sarcomeric architecture in CMs. ACTN4 influences fractional shortening at the cell level and contractility at the tissue level. Changes in ventricular ACTN4 levels are associated with remodeling and may influence clinical outcomes related to heart failure.","PeriodicalId":10147,"journal":{"name":"Circulation research","volume":"100 1","pages":""},"PeriodicalIF":20.1,"publicationDate":"2025-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145545085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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