Pub Date : 2025-02-18DOI: 10.1016/j.yjmcc.2025.01.011
Alexa Corker , Miguel Troncoso , Maya Learmonth , Philip Broughton , Sara J. Sidles , Ryan Kelly , Shaoni Dasgupta , Thomas Dempster , Kim Vu , Amber Hazzard , An Van Laer , Rachel D. Penrod , Jeffery A. Jones , Amy D. Bradshaw , Michael R. Zile , Amanda C. LaRue , Kristine Y. DeLeon-Pennell
Post-traumatic stress disorder (PTSD) is a disabling psychological disorder characterized by chronic symptoms of intrusiveness, avoidance, and hyperarousal after a traumatic event. Retrospective studies have indicated PTSD increases the risk for cardiovascular disease (CVD) including arrhythmia, hypertension, and myocardial infarction. The goal of this study was to: 1) use a murine model of cued fear conditioning (inescapable foot shock, IFS) to develop a scoring method to distinguish a PTSD-like phenotype, and 2) use this model system to characterize the cardiac phenotype and function in mice with extreme PTSD-like behaviors. We compared 3 groups, controls, non-responders (NR), and PTSD-like mice at 2 time points [4-weeks and 8-weeks post-IFS] to compare left ventricular structure and function. Assessment of cardiac function showed both male and female PTSD-like mice had increased isovolumetric relaxation time at 8-weeks post-IFS, whereas only females demonstrated increases in E/e’, left atrial diameter, and decreased ejection fraction compared to control mice. Female PTSD-like mice also demonstrated increased interstitial fibrosis through picrosirius red staining and increased expression of fibrotic genes including Col3a1 and Lox. Overall, our data indicated that mice displaying behavioral characteristics associated with PTSD present with sex-dependent diastolic dysfunction likely due, at least in part, to an activation of cardiac fibrosis.
{"title":"Mouse model of post-traumatic stress disorder negatively impacts cardiac homeostasis","authors":"Alexa Corker , Miguel Troncoso , Maya Learmonth , Philip Broughton , Sara J. Sidles , Ryan Kelly , Shaoni Dasgupta , Thomas Dempster , Kim Vu , Amber Hazzard , An Van Laer , Rachel D. Penrod , Jeffery A. Jones , Amy D. Bradshaw , Michael R. Zile , Amanda C. LaRue , Kristine Y. DeLeon-Pennell","doi":"10.1016/j.yjmcc.2025.01.011","DOIUrl":"10.1016/j.yjmcc.2025.01.011","url":null,"abstract":"<div><div>Post-traumatic stress disorder (PTSD) is a disabling psychological disorder characterized by chronic symptoms of intrusiveness, avoidance, and hyperarousal after a traumatic event. Retrospective studies have indicated PTSD increases the risk for cardiovascular disease (CVD) including arrhythmia, hypertension, and myocardial infarction. The goal of this study was to: 1) use a murine model of cued fear conditioning (inescapable foot shock, IFS) to develop a scoring method to distinguish a PTSD-like phenotype, and 2) use this model system to characterize the cardiac phenotype and function in mice with extreme PTSD-like behaviors. We compared 3 groups, controls, non-responders (NR), and PTSD-like mice at 2 time points [4-weeks and 8-weeks post-IFS] to compare left ventricular structure and function. Assessment of cardiac function showed both male and female PTSD-like mice had increased isovolumetric relaxation time at 8-weeks post-IFS, whereas only females demonstrated increases in E/e’, left atrial diameter, and decreased ejection fraction compared to control mice. Female PTSD-like mice also demonstrated increased interstitial fibrosis through picrosirius red staining and increased expression of fibrotic genes including <em>Col3a1</em> and <em>Lox</em>. Overall, our data indicated that mice displaying behavioral characteristics associated with PTSD present with sex-dependent diastolic dysfunction likely due, at least in part, to an activation of cardiac fibrosis.</div></div>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":"201 ","pages":"Pages 32-43"},"PeriodicalIF":4.9,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143429833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-13DOI: 10.1016/j.yjmcc.2025.02.003
Zehao Yao, Lina Bai, Yu Nie
Measuring cardiomyocyte nuclear ploidy is crucial for evaluating broader aspects of cardiac development, function, and disease progression. Fluorescence in situ hybridization (FISH) remains the gold standard for ploidy identification; however, its application in cardiomyocytes is hindered by their unique cellular complexities. Here, we describe a detailed cardiomyocyte-specific FISH (cardioFISH) protocol. CardioFISH incorporates a tailored enzymatic digestion strategy to enhances nuclear accessibility while preserving cellular integrity and minimizing sarcomere-derived autofluorescence. Additionally, we introduce a 3D nuclear visualization framework for comprehensive cardioFISH signal analysis, addressing the limitations imposed by the large nuclear dimensions of cardiomyocytes, where signals are frequently distributed across multiple imaging planes. This two-day cardioFISH protocol is applicable to various stages of cardiomyocyte development and provides a powerful tool for advancing studies of cardiomyocyte ploidy.
{"title":"Fluorescence in situ hybridization protocol for cardiomyocytes.","authors":"Zehao Yao, Lina Bai, Yu Nie","doi":"10.1016/j.yjmcc.2025.02.003","DOIUrl":"https://doi.org/10.1016/j.yjmcc.2025.02.003","url":null,"abstract":"<p><p>Measuring cardiomyocyte nuclear ploidy is crucial for evaluating broader aspects of cardiac development, function, and disease progression. Fluorescence in situ hybridization (FISH) remains the gold standard for ploidy identification; however, its application in cardiomyocytes is hindered by their unique cellular complexities. Here, we describe a detailed cardiomyocyte-specific FISH (cardioFISH) protocol. CardioFISH incorporates a tailored enzymatic digestion strategy to enhances nuclear accessibility while preserving cellular integrity and minimizing sarcomere-derived autofluorescence. Additionally, we introduce a 3D nuclear visualization framework for comprehensive cardioFISH signal analysis, addressing the limitations imposed by the large nuclear dimensions of cardiomyocytes, where signals are frequently distributed across multiple imaging planes. This two-day cardioFISH protocol is applicable to various stages of cardiomyocyte development and provides a powerful tool for advancing studies of cardiomyocyte ploidy.</p>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143425624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Overconsumption of fructose has been linked to the development of systemic metabolic and cardiac diseases, yet few studies have focused on the link between cardiac fructose metabolism and the development of heart disease. Low-oxygen complex exercise is considered an effective means of treating and preventing metabolic diseases and improving cardiac function, however, it is unclear, the link between low-oxygen complex exercise and high-fructose-induced heart disease. Therefore, the aim of this study was to investigate the effect of hypoxic complex exercise on heart disease on a high fructose diet. The results of the study found that hypoxic compound exercise improved the upregulation of inflammatory factor Upd3 and systemic fat accumulation in the heart induced by high fructose diet by inhibiting the expression of KHK gene in the heart; and it improved the impaired cardiac rhythmic function and pumping function, improved the disorder of myofilament fiber arrangement, reduced the level of cardiac oxidative stress, and reduced cardiac collagen deposition. In addition, cardiac KHK-specific knockdown had the same effect on high fructose diet hearts. Compared with single KHK cardiac-specific knockdown or hypoxic combination exercise, hypoxic combination exercise combined with KHK cardiac-specific knockdown was superior in improving the high-fructose diet-induced increase in arrhythmia index, systolic and diastolic dysfunction, and decrease in fractional shortening. Therefore, we conclude that hypoxic complex exercise improved high-fructose diet-induced cardiac rhythmic function and pumping dysfunction by reducing KHK expression.
{"title":"Hypoxic compound exercise improves cardiac function in Drosophila high fructose diet via KHK.","authors":"Xu Ping, Qiufang Li, Meng Ding, Zhengwen Yu, Qin Yi, Yuepeng Li, Wenzhi Gu, Ping Zhang, Zike Zhang, Lan Zheng","doi":"10.1016/j.yjmcc.2025.02.005","DOIUrl":"https://doi.org/10.1016/j.yjmcc.2025.02.005","url":null,"abstract":"<p><p>Overconsumption of fructose has been linked to the development of systemic metabolic and cardiac diseases, yet few studies have focused on the link between cardiac fructose metabolism and the development of heart disease. Low-oxygen complex exercise is considered an effective means of treating and preventing metabolic diseases and improving cardiac function, however, it is unclear, the link between low-oxygen complex exercise and high-fructose-induced heart disease. Therefore, the aim of this study was to investigate the effect of hypoxic complex exercise on heart disease on a high fructose diet. The results of the study found that hypoxic compound exercise improved the upregulation of inflammatory factor Upd3 and systemic fat accumulation in the heart induced by high fructose diet by inhibiting the expression of KHK gene in the heart; and it improved the impaired cardiac rhythmic function and pumping function, improved the disorder of myofilament fiber arrangement, reduced the level of cardiac oxidative stress, and reduced cardiac collagen deposition. In addition, cardiac KHK-specific knockdown had the same effect on high fructose diet hearts. Compared with single KHK cardiac-specific knockdown or hypoxic combination exercise, hypoxic combination exercise combined with KHK cardiac-specific knockdown was superior in improving the high-fructose diet-induced increase in arrhythmia index, systolic and diastolic dysfunction, and decrease in fractional shortening. Therefore, we conclude that hypoxic complex exercise improved high-fructose diet-induced cardiac rhythmic function and pumping dysfunction by reducing KHK expression.</p>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143425625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background: Thoracic aortic aneurysm and dissection (TAAD) is a life-threatening cardiovascular event characterized by high mortality rates. Previous studies have shown that matrix metalloproteinases 19 (MMP19) was involved in TAAD formation, while the detailed role of MMP19 in TAAD pathogenesis and underlying mechanism remain unclear.
Methods: To investigate the role of MMP19 in the progression of TAAD, we generated global Mmp19 knockout mice, as well as VSMCs (vascular smooth muscle cells)-specific Mmp19 knockdown mice, and established a BAPN-induced TAAD model. To elucidate the signaling pathways modulated by Aggrecan, we employed an adeno-associated virus serotype 9 (AAV9) vector encoding Acan short hairpin RNA (shRNA) for VSMC-specific knockdown of Acan. Ultimately, we injected an AAV vector encoding VSMC-specific Mmp19 into BAPN-induced TAAD mice to assess whether MMP19 can mitigate the development of TAAD.
Results: Our findings revealed elevated mRNA and protein levels of MMP19 in the aortas of both TAAD mice and patients. The systemic ablation of Mmp19, as well as VSMC-specific Mmp19 knockdown, significantly exacerbated BAPN-induced progressive TAAD, and TAAD-related cardiovascular remodeling. Mmp19 deficiency resulted in the accumulation of Acan, but not Vcan, within the aorta, driving the phenotypic switch of VSMCs from contractile to synthetic state through activting Wnt/β-catenin signaling pathway. The selective inhibitor of Wnt/β-catenin signaling, MASB, was effective in reversing the dedifferentiation of VSMCs induced by aggrecan accumulation. Notably, the specific knockdown of Acan in VSMCs restored the contractile phenotype of VSMCs and inhibited Wnt/β-catenin signaling, thereby alleviating BAPN-induced TAAD in Mmp19-/- mice. Additionally, VSMC-specific complementation of MMP19 also alleviated the progressive TAAD phenotype in Mmp19-/- mice.
Conclusions: The study underscores that MMP19 deficiency exacerbates TAAD by promoting Acan aggregation and destroying the homeostasis of VSMCs by activating Wnt/β-catenin signaling pathway. These results posit MMP19 as a promising novel therapeutic target for TAAD intervention.
{"title":"MMP19 in vascular smooth muscle cells protects against thoracic aortic aneurysm and dissection via the MMP19/Aggrecan/Wnt/β-catenin axis.","authors":"Baihui Ma, Qingyi Zeng, Fangfang Yang, Hang Yang, Wenke Li, Rui Fu, Zeyu Cai, Guoyan Zhu, Chang Shu, Mingyao Luo, Zhou Zhou","doi":"10.1016/j.yjmcc.2025.02.004","DOIUrl":"https://doi.org/10.1016/j.yjmcc.2025.02.004","url":null,"abstract":"<p><strong>Background: </strong>Thoracic aortic aneurysm and dissection (TAAD) is a life-threatening cardiovascular event characterized by high mortality rates. Previous studies have shown that matrix metalloproteinases 19 (MMP19) was involved in TAAD formation, while the detailed role of MMP19 in TAAD pathogenesis and underlying mechanism remain unclear.</p><p><strong>Methods: </strong>To investigate the role of MMP19 in the progression of TAAD, we generated global Mmp19 knockout mice, as well as VSMCs (vascular smooth muscle cells)-specific Mmp19 knockdown mice, and established a BAPN-induced TAAD model. To elucidate the signaling pathways modulated by Aggrecan, we employed an adeno-associated virus serotype 9 (AAV9) vector encoding Acan short hairpin RNA (shRNA) for VSMC-specific knockdown of Acan. Ultimately, we injected an AAV vector encoding VSMC-specific Mmp19 into BAPN-induced TAAD mice to assess whether MMP19 can mitigate the development of TAAD.</p><p><strong>Results: </strong>Our findings revealed elevated mRNA and protein levels of MMP19 in the aortas of both TAAD mice and patients. The systemic ablation of Mmp19, as well as VSMC-specific Mmp19 knockdown, significantly exacerbated BAPN-induced progressive TAAD, and TAAD-related cardiovascular remodeling. Mmp19 deficiency resulted in the accumulation of Acan, but not Vcan, within the aorta, driving the phenotypic switch of VSMCs from contractile to synthetic state through activting Wnt/β-catenin signaling pathway. The selective inhibitor of Wnt/β-catenin signaling, MASB, was effective in reversing the dedifferentiation of VSMCs induced by aggrecan accumulation. Notably, the specific knockdown of Acan in VSMCs restored the contractile phenotype of VSMCs and inhibited Wnt/β-catenin signaling, thereby alleviating BAPN-induced TAAD in Mmp19<sup>-/-</sup> mice. Additionally, VSMC-specific complementation of MMP19 also alleviated the progressive TAAD phenotype in Mmp19<sup>-/-</sup> mice.</p><p><strong>Conclusions: </strong>The study underscores that MMP19 deficiency exacerbates TAAD by promoting Acan aggregation and destroying the homeostasis of VSMCs by activating Wnt/β-catenin signaling pathway. These results posit MMP19 as a promising novel therapeutic target for TAAD intervention.</p>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143425626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-08DOI: 10.1016/j.yjmcc.2025.02.001
R.A. Di Mattía , D. Gallo , S. Ciarrocchi , L.A. Gonano , P.G. Blanco , C.A. Valverde , E.L. Portiansky , L.M. Sommese , K. Toischer , F. Bleckwedel , L.C. Zelarayán , E.A. Aiello , A. Orlowski
<div><h3>Introduction</h3><div>IRBIT, also known as Ahcyl1, is an IP3 receptor (IP3R)-binding protein released with IP3 and was first described as a competitive inhibitor of the mentioned receptor. Studies have shown that overexpression of IP3Rs is associated with cardiac hypertrophy in both human and animal models. Given that IP3Rs play a role in cardiac hypertrophy, IRBIT may also be involved in this condition.</div></div><div><h3>Aim</h3><div>Although IRBIT heart expression has been reported, its function in cardiac tissues remains unclear. Thus, we aimed to study the cardiac outcomes of up-and downregulation of IRBIT to establish its pathophysiological role.</div></div><div><h3>Methods and results</h3><div>We found that IRBIT is expressed in mouse ventricles and atria, fibroblasts and cardiomyocytes isolated from neonatal mice, and in the myoblast cell line H9c2. Mice with transverse aortic constriction showed a significant increase in both the mRNA and protein expression of IRBIT. Furthermore, we described the differential expression of IRBIT in human myocardial samples of dilated and ischemic cardiomyopathy. IRBIT cardiac overexpression in mice using an adenoassociated virus (AAV9) at two different time points (neonatal mice, day 4 and adult mice, 3 months) resulted in the development of cardiac hypertrophy with impaired systolic function by four months of age. A decrease in the mRNA levels of the IP3 receptor was also observed in both models. Isolated myocytes from the IRBIT-overexpressing neonatal model showed a significantly decreased Ca<sup>2+</sup> transient amplitude and slower rise of the global Ca<sup>2+</sup> transient, without changes in sarcoplasmic reticulum (SR) Ca<sup>2+</sup> content or spontaneous Ca<sup>2+</sup> wave frequency. However, the velocity of Ca<sup>2+</sup> wave propagation was reduced. Moreover, we found that the dyssynchrony index (DI) is significantly increased under IRBIT overexpression. Nuclear Ca<sup>2+</sup> dynamics were assessed, showing no significant changes, but IRBIT overexpression reduced the number of nuclear envelope invaginations. In addition, reducing IRBIT expression using AAV9-shRNA did not result in any changes in the heart morphometric parameters.</div></div><div><h3>Conclusion</h3><div>Our study describes for the first time that IRBIT plays a critical role in the pathophysiology of the heart. Our findings demonstrate that IRBIT overexpression disrupts Ca<sup>2+</sup> signaling, contributing to hypertrophic remodeling and impaired cardiac function. The altered wave propagation, the increase in DI and the decrease of the rate of the Ca<sup>2+</sup> transient suggests that IRBIT influences Ca<sup>2+</sup> − induced Ca<sup>2+</sup> release. This study provides the first evidence linking IRBIT to pathological cardiac remodeling and Ca<sup>2+</sup> handling dysregulation. Although significant progress has been made, further research is required to better understand the cardiovascular function of
{"title":"Cardiac hypertrophy induced by overexpression of IP3-released inositol 1, 4, 5-trisphosphate receptor-binding protein (IRBIT)","authors":"R.A. Di Mattía , D. Gallo , S. Ciarrocchi , L.A. Gonano , P.G. Blanco , C.A. Valverde , E.L. Portiansky , L.M. Sommese , K. Toischer , F. Bleckwedel , L.C. Zelarayán , E.A. Aiello , A. Orlowski","doi":"10.1016/j.yjmcc.2025.02.001","DOIUrl":"10.1016/j.yjmcc.2025.02.001","url":null,"abstract":"<div><h3>Introduction</h3><div>IRBIT, also known as Ahcyl1, is an IP3 receptor (IP3R)-binding protein released with IP3 and was first described as a competitive inhibitor of the mentioned receptor. Studies have shown that overexpression of IP3Rs is associated with cardiac hypertrophy in both human and animal models. Given that IP3Rs play a role in cardiac hypertrophy, IRBIT may also be involved in this condition.</div></div><div><h3>Aim</h3><div>Although IRBIT heart expression has been reported, its function in cardiac tissues remains unclear. Thus, we aimed to study the cardiac outcomes of up-and downregulation of IRBIT to establish its pathophysiological role.</div></div><div><h3>Methods and results</h3><div>We found that IRBIT is expressed in mouse ventricles and atria, fibroblasts and cardiomyocytes isolated from neonatal mice, and in the myoblast cell line H9c2. Mice with transverse aortic constriction showed a significant increase in both the mRNA and protein expression of IRBIT. Furthermore, we described the differential expression of IRBIT in human myocardial samples of dilated and ischemic cardiomyopathy. IRBIT cardiac overexpression in mice using an adenoassociated virus (AAV9) at two different time points (neonatal mice, day 4 and adult mice, 3 months) resulted in the development of cardiac hypertrophy with impaired systolic function by four months of age. A decrease in the mRNA levels of the IP3 receptor was also observed in both models. Isolated myocytes from the IRBIT-overexpressing neonatal model showed a significantly decreased Ca<sup>2+</sup> transient amplitude and slower rise of the global Ca<sup>2+</sup> transient, without changes in sarcoplasmic reticulum (SR) Ca<sup>2+</sup> content or spontaneous Ca<sup>2+</sup> wave frequency. However, the velocity of Ca<sup>2+</sup> wave propagation was reduced. Moreover, we found that the dyssynchrony index (DI) is significantly increased under IRBIT overexpression. Nuclear Ca<sup>2+</sup> dynamics were assessed, showing no significant changes, but IRBIT overexpression reduced the number of nuclear envelope invaginations. In addition, reducing IRBIT expression using AAV9-shRNA did not result in any changes in the heart morphometric parameters.</div></div><div><h3>Conclusion</h3><div>Our study describes for the first time that IRBIT plays a critical role in the pathophysiology of the heart. Our findings demonstrate that IRBIT overexpression disrupts Ca<sup>2+</sup> signaling, contributing to hypertrophic remodeling and impaired cardiac function. The altered wave propagation, the increase in DI and the decrease of the rate of the Ca<sup>2+</sup> transient suggests that IRBIT influences Ca<sup>2+</sup> − induced Ca<sup>2+</sup> release. This study provides the first evidence linking IRBIT to pathological cardiac remodeling and Ca<sup>2+</sup> handling dysregulation. Although significant progress has been made, further research is required to better understand the cardiovascular function of ","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":"201 ","pages":"Pages 1-15"},"PeriodicalIF":4.9,"publicationDate":"2025-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143391047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-07DOI: 10.1016/j.yjmcc.2025.02.002
Angela C. Greenman, Rachel L. Sadler, Samantha P. Harris
Cardiac myosin binding protein-C (cMyBP-C) is a sarcomere regulatory protein consisting of 11 well-folded immunoglobulin-like (Ig-like) and fibronectin type-III domains with the individual domains numbered C0-C10. Despite progress in understanding the functions of the N′ and C′-terminal ends of the protein, our understanding of the functional effects of the middle domains (C3-C4-C5-C6-C7) is still limited. Here we aimed to determine the functional significance of the middle domains by replacing endogenous cMyBP-C with recombinant proteins with and without the middle domains using our “cut and paste” SpyC3 mouse model. Specifically, we deleted domains C3-C7 or substituted these domains with unrelated Ig-like domains from titin to behave as inert “spacer” domains. Replacement with the spacer constructs resulted in a significant increase in myofilament calcium sensitivity, an almost instantaneous redevelopment of tension after a slack re-stretch protocol, and altered stretch activation responses, suggesting that the middle domains are functionally relevant and normally exert inhibitory effects on force development. We also investigated the significance of a potentially flexible linker between domains C4 and C5 and a unique 28 amino acid loop insertion in C5. Whereas deletion of the C5 loop had no effect on force, deletion of the linker between C4 and C5 had comparable effects to deletion of domains C3-C7. Taken together, these data indicate that the middle domains play an important role in limiting the activating effects of the C0-C2 domains and that the C4C5 linker contributes to these effects.
{"title":"Autoinhibition of cMyBP-C by its middle domains","authors":"Angela C. Greenman, Rachel L. Sadler, Samantha P. Harris","doi":"10.1016/j.yjmcc.2025.02.002","DOIUrl":"10.1016/j.yjmcc.2025.02.002","url":null,"abstract":"<div><div>Cardiac myosin binding protein-C (cMyBP-C) is a sarcomere regulatory protein consisting of 11 well-folded immunoglobulin-like (Ig-like) and fibronectin type-III domains with the individual domains numbered C0-C10. Despite progress in understanding the functions of the N′ and C′-terminal ends of the protein, our understanding of the functional effects of the middle domains (C3-C4-C5-C6-C7) is still limited. Here we aimed to determine the functional significance of the middle domains by replacing endogenous cMyBP-C with recombinant proteins with and without the middle domains using our “cut and paste” SpyC3 mouse model. Specifically, we deleted domains C3-C7 or substituted these domains with unrelated Ig-like domains from titin to behave as inert “spacer” domains. Replacement with the spacer constructs resulted in a significant increase in myofilament calcium sensitivity, an almost instantaneous redevelopment of tension after a slack re-stretch protocol, and altered stretch activation responses, suggesting that the middle domains are functionally relevant and normally exert inhibitory effects on force development. We also investigated the significance of a potentially flexible linker between domains C4 and C5 and a unique 28 amino acid loop insertion in C5. Whereas deletion of the C5 loop had no effect on force, deletion of the linker between C4 and C5 had comparable effects to deletion of domains C3-C7. Taken together, these data indicate that the middle domains play an important role in limiting the activating effects of the C0-C2 domains and that the C4C5 linker contributes to these effects.</div></div>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":"200 ","pages":"Pages 82-92"},"PeriodicalIF":4.9,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143382708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-03DOI: 10.1016/j.yjmcc.2025.01.010
Maria Uscategui Calderon , Maria L. Spaeth , Marissa Granitto , Brittany A. Gonzalez , Matthew T. Weirauch , Leah C. Kottyan , Katherine E. Yutzey
Cardiomyocytes and cardiac fibroblasts undergo coordinated maturation after birth, and cardiac fibroblasts are required for postnatal cardiomyocyte maturation in mice. Here, we investigate the role of cardiac fibroblast-expressed Growth Differentiation Factor 10 (GDF10) in postnatal heart development. In neonatal mice, Gdf10 is expressed specifically in cardiac fibroblasts, with its highest expression coincident with the onset of cardiomyocyte cell cycle arrest and transition to hypertrophic growth. In neonatal rat ventricular myocyte (NRVM) cultures, GDF10 treatment promotes cardiomyocyte maturation indicated by increased binucleation, downregulation of cell cycle progression genes, and upregulation of cell cycle inhibitor genes. GDF10 treatment leads to an increase in cardiomyocyte cell size, together with increased expression of mature sarcomeric protein isoforms and decreased expression of fetal cardiac genes. RNAsequencing of GDF10-treated NRVM shows an increase in the expression of genes related to myocardial maturation, including upregulation of sodium and potassium channel genes. In vivo, loss of Gdf10 leads to a delay in myocardial maturation indicated by decreased cardiomyocyte cell size and binucleation, as well as increased mitotic activity, at postnatal (P) day 7. Further, induction of mature sarcomeric protein isoform gene expression is delayed, and expression of cell cycle progression genes is prolonged. However, by P10, indicators of cardiomyocyte maturation and mitotic activity are normalized in Gdf10-null hearts relative to controls. Together, these results implicate GDF10 as a novel crosstalk mediator between cardiomyocytes and cardiac fibroblasts, which is required for appropriate timing of cardiomyocyte maturation steps including binucleation, hypertrophy, mature sarcomeric isoform gene expression, and cell cycle arrest in the postnatal period.
{"title":"GDF10 promotes rodent cardiomyocyte maturation during the postnatal period","authors":"Maria Uscategui Calderon , Maria L. Spaeth , Marissa Granitto , Brittany A. Gonzalez , Matthew T. Weirauch , Leah C. Kottyan , Katherine E. Yutzey","doi":"10.1016/j.yjmcc.2025.01.010","DOIUrl":"10.1016/j.yjmcc.2025.01.010","url":null,"abstract":"<div><div>Cardiomyocytes and cardiac fibroblasts undergo coordinated maturation after birth, and cardiac fibroblasts are required for postnatal cardiomyocyte maturation in mice. Here, we investigate the role of cardiac fibroblast-expressed Growth Differentiation Factor 10 (GDF10) in postnatal heart development. In neonatal mice, <em>Gdf10</em> is expressed specifically in cardiac fibroblasts, with its highest expression coincident with the onset of cardiomyocyte cell cycle arrest and transition to hypertrophic growth. In neonatal rat ventricular myocyte (NRVM) cultures, GDF10 treatment promotes cardiomyocyte maturation indicated by increased binucleation, downregulation of cell cycle progression genes, and upregulation of cell cycle inhibitor genes. GDF10 treatment leads to an increase in cardiomyocyte cell size, together with increased expression of mature sarcomeric protein isoforms and decreased expression of fetal cardiac genes. RNAsequencing of GDF10-treated NRVM shows an increase in the expression of genes related to myocardial maturation, including upregulation of sodium and potassium channel genes. <em>In vivo</em>, loss of <em>Gdf10</em> leads to a delay in myocardial maturation indicated by decreased cardiomyocyte cell size and binucleation, as well as increased mitotic activity, at postnatal (P) day 7. Further, induction of mature sarcomeric protein isoform gene expression is delayed, and expression of cell cycle progression genes is prolonged. However, by P10, indicators of cardiomyocyte maturation and mitotic activity are normalized in <em>Gdf10</em>-null hearts relative to controls. Together, these results implicate GDF10 as a novel crosstalk mediator between cardiomyocytes and cardiac fibroblasts, which is required for appropriate timing of cardiomyocyte maturation steps including binucleation, hypertrophy, mature sarcomeric isoform gene expression, and cell cycle arrest in the postnatal period.</div></div>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":"201 ","pages":"Pages 16-31"},"PeriodicalIF":4.9,"publicationDate":"2025-02-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143255906","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.yjmcc.2024.12.007
Jarkko P. Hytönen , Juho Pajula , Paavo Halonen , Jouni Taavitsainen , Antti Kuivanen , Santeri Tarvainen , Minja Heikkilä , Petri Mäkinen , Arto Koistinen , Johanna P. Laakkonen , Juha Hartikainen , Seppo Ylä-Herttuala
Background
Coronary stenting operations have become the main option for the treatment of coronary heart disease. Vessel recovery after stenting has emerged as a critical factor in reducing possible complications. In this study, we evaluated the feasibility, safety and efficacy of locally administered intraluminal gene therapy delivered using a specialized infusion balloon catheter.
Methods
Sixteen pigs received bare metal stents (BMS) in the left circumflex coronary artery and drug-eluting stents (DES) in the right coronary artery. Adenoviral (Ad) gene transfers (dose 1,5e10 pfu) of VEGF-A and LacZ were performed with a ClearWay™ RX infusion balloon catheter on both stents. In vivo imaging included angiography, OCT and angioscopy. Tissue samples were collected for analyses at day 14 and studied using scanning electron microscopy and multiphoton microscopy.
Results
AdVEGF-A accelerated re-endothelialization in the BMS compared to the other groups. The highest restenosis was in the BMS AdLacZ group. DES groups had statistically significant reduced maximum stenosis compared to BMS AdLacZ. No major in-stent thrombosis events were detected. Ex vivo imaging showed that intraarterial imaging is not very accurate in the detection of endothelial layer. Biodistribution of the Ad vector and measured safety parameters (off-target tissues and blood tests) did not show any major safety concerns.
Conclusion
Adenoviral gene transfer using the ClearWay™ RX catheter was feasible and safe. AdVEGF-A accelerated re-endothelization in BMS. However, DES showed better outcomes in the short-term regarding restenosis and do not benefit from intraluminal AdVEGF-A gene transfer.
{"title":"Endothelialization of coronary stents after intra-luminal adenoviral VEGF-A gene transfer in a preclinical porcine restenosis model – Studies with optical coherence tomography, angioscopy, multiphoton and scanning electron microscopy","authors":"Jarkko P. Hytönen , Juho Pajula , Paavo Halonen , Jouni Taavitsainen , Antti Kuivanen , Santeri Tarvainen , Minja Heikkilä , Petri Mäkinen , Arto Koistinen , Johanna P. Laakkonen , Juha Hartikainen , Seppo Ylä-Herttuala","doi":"10.1016/j.yjmcc.2024.12.007","DOIUrl":"10.1016/j.yjmcc.2024.12.007","url":null,"abstract":"<div><h3>Background</h3><div>Coronary stenting operations have become the main option for the treatment of coronary heart disease. Vessel recovery after stenting has emerged as a critical factor in reducing possible complications. In this study, we evaluated the feasibility, safety and efficacy of locally administered intraluminal gene therapy delivered using a specialized infusion balloon catheter.</div></div><div><h3>Methods</h3><div>Sixteen pigs received bare metal stents (BMS) in the left circumflex coronary artery and drug-eluting stents (DES) in the right coronary artery. Adenoviral (Ad) gene transfers (dose 1,5e10 pfu) of VEGF-A and LacZ were performed with a ClearWay™ RX infusion balloon catheter on both stents. In vivo imaging included angiography, OCT and angioscopy. Tissue samples were collected for analyses at day 14 and studied using scanning electron microscopy and multiphoton microscopy.</div></div><div><h3>Results</h3><div>AdVEGF-A accelerated re-endothelialization in the BMS compared to the other groups. The highest restenosis was in the BMS AdLacZ group. DES groups had statistically significant reduced maximum stenosis compared to BMS AdLacZ. No major in-stent thrombosis events were detected. Ex vivo imaging showed that intraarterial imaging is not very accurate in the detection of endothelial layer. Biodistribution of the Ad vector and measured safety parameters (off-target tissues and blood tests) did not show any major safety concerns.</div></div><div><h3>Conclusion</h3><div>Adenoviral gene transfer using the ClearWay™ RX catheter was feasible and safe. AdVEGF-A accelerated re-endothelization in BMS. However, DES showed better outcomes in the short-term regarding restenosis and do not benefit from intraluminal AdVEGF-A gene transfer.</div></div>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":"199 ","pages":"Pages 118-125"},"PeriodicalIF":4.9,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142927022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.yjmcc.2024.12.008
Ruzzell C. Flores, Rachel Yaffe, Munashe M. Nhunzwi, Huong Nguyen, Inna Rabinovich-Nikitin
Cardiovascular disease (CVD) is the leading cause of death for women worldwide. One of the risk factors for CVD in women is complications during pregnancy. Pregnancy complications include a wide arena of pathologies, including hypertension, preeclampsia, gestational diabetes, preterm delivery and miscarriage. Interestingly, increased evidence in recent years highlights a novel link between maternal shift work during pregnancy and increased risk for pregnancy complications, specifically hypertension and diabetes, while knowledge on other CVDs, such heart failure, atherosclerosis, ischemic heart disease, and stroke in pregnant shift working mothers is still scarce. Notably, shift work during pregnancy results in significant changes to the circadian rhythm of both the mother and fetus, therefore, engaging into shift work during pregnancy may adversely affect the cardiovascular health of both the mother and offspring, and carry into adulthood. Herein, we highlight the novel relationship between maternal shift work during pregnancy and the increased risk for pregnancy complications that may increase risk for CVD later in life. Furthermore, we provide mechanistic insights of the hemodynamic processes that are disrupted in response to maternal shift work and may explain the increased risk for cardiovascular disease.
Understanding how shift work during pregnancy influences the prevalence for heart disease is of paramount clinical importance for minimizing the risk for cardiovascular disease for both the mother and offspring.
{"title":"Maternal shift work during pregnancy and cardiovascular health impacts on mother and offspring","authors":"Ruzzell C. Flores, Rachel Yaffe, Munashe M. Nhunzwi, Huong Nguyen, Inna Rabinovich-Nikitin","doi":"10.1016/j.yjmcc.2024.12.008","DOIUrl":"10.1016/j.yjmcc.2024.12.008","url":null,"abstract":"<div><div>Cardiovascular disease (CVD) is the leading cause of death for women worldwide. One of the risk factors for CVD in women is complications during pregnancy. Pregnancy complications include a wide arena of pathologies, including hypertension, preeclampsia, gestational diabetes, preterm delivery and miscarriage. Interestingly, increased evidence in recent years highlights a novel link between maternal shift work during pregnancy and increased risk for pregnancy complications, specifically hypertension and diabetes, while knowledge on other CVDs, such heart failure, atherosclerosis, ischemic heart disease, and stroke in pregnant shift working mothers is still scarce. Notably, shift work during pregnancy results in significant changes to the circadian rhythm of both the mother and fetus, therefore, engaging into shift work during pregnancy may adversely affect the cardiovascular health of both the mother and offspring, and carry into adulthood. Herein, we highlight the novel relationship between maternal shift work during pregnancy and the increased risk for pregnancy complications that may increase risk for CVD later in life. Furthermore, we provide mechanistic insights of the hemodynamic processes that are disrupted in response to maternal shift work and may explain the increased risk for cardiovascular disease.</div><div>Understanding how shift work during pregnancy influences the prevalence for heart disease is of paramount clinical importance for minimizing the risk for cardiovascular disease for both the mother and offspring.</div></div>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":"199 ","pages":"Pages 126-132"},"PeriodicalIF":4.9,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142927082","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.yjmcc.2024.12.001
Sharanya S. Bettadapura , William D. Todd , Graham R. McGinnis , Danielle R. Bruns
The age of the U.S. population is increasing alongside a growing burden of age-related cardiovascular disease. Circadian rhythms are critical for human health and are disrupted with aging and cardiovascular disease. The goal of the present review is to summarize how cardiac circadian rhythms change with age and how this might contribute to the increasing burden of age-associated heart disease. Further, we will review what is known about interventions to slow aging and whether they impact cardiac clock function, as well as whether time-of-day or chronotherapy may improve cardiac function with age. Although much remains to be understood about the circadian biology of cardiac aging, we propose that altered circadian clock output should be considered a hallmark of aging and that efforts to fix the clock are warranted for healthy cardiac aging.
{"title":"Circadian biology of cardiac aging","authors":"Sharanya S. Bettadapura , William D. Todd , Graham R. McGinnis , Danielle R. Bruns","doi":"10.1016/j.yjmcc.2024.12.001","DOIUrl":"10.1016/j.yjmcc.2024.12.001","url":null,"abstract":"<div><div>The age of the U.S. population is increasing alongside a growing burden of age-related cardiovascular disease. Circadian rhythms are critical for human health and are disrupted with aging and cardiovascular disease. The goal of the present review is to summarize how cardiac circadian rhythms change with age and how this might contribute to the increasing burden of age-associated heart disease. Further, we will review what is known about interventions to slow aging and whether they impact cardiac clock function, as well as whether time-of-day or chronotherapy may improve cardiac function with age. Although much remains to be understood about the circadian biology of cardiac aging, we propose that altered circadian clock output should be considered a hallmark of aging and that efforts to fix the clock are warranted for healthy cardiac aging.</div></div>","PeriodicalId":16402,"journal":{"name":"Journal of molecular and cellular cardiology","volume":"199 ","pages":"Pages 95-103"},"PeriodicalIF":4.9,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142927093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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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