Long Noncoding RNA Cardiac Physiological Hypertrophy–Associated Regulator Induces Cardiac Physiological Hypertrophy and Promotes Functional Recovery After Myocardial Ischemia-Reperfusion Injury

Abstract

Background: Exercise training's benefits in cardiovascular system have been well accepted, however, the underlying mechanism remains to be explored. Here, we report the initial functional characterization of an exercise-induced cardiac physiological hypertrophy associated novel lncRNA. Methods: Using lncRNA microarray profiling, we identified lncRNAs in contributing the modulation of exercise-induced cardiac growth that we termed Cardiac Physiological hypertrophy associated regulator (CPhar). Mice with Adeno-associated virus serotype 9 (AAV9) driving CPhar overexpression and knockdown were used in in-vivo experiments. Swim training was used to induce physiological cardiac hypertrophy in mice and ischemia reperfusion injury (IR/I) surgery was conducted to investigate the protective effects of CPhar in mice. To investigate the mechanisms of CPhar's function, we performed various analysis including RTqPCR, western blot, histology, cardiac function (by echocardiography), functional rescue experiments, mass spectrometry, in vitro RNA transcription, RNA pull down, RNA immunoprecipitation, chromatin immunoprecipitation assay, luciferase reporter assay, and coimmunoprecipitation assays. Results: We screened the lncRNAs in contributing the modulation of exercise-induced cardiac growth via lncRNA microarray profiling and found that CPhar was increased with exercise and was necessary for exercise-induced physiological cardiac growth. Gain- and loss- of function of CPhar regulated the expression of proliferation markers, hypertrophy, and apoptosis in cultured neonatal mouse cardiomyocytes (NMCMs). Overexpression of CPhar prevented myocardial ischemia reperfusion injury and cardiac dysfunction in vivo . We identified DDX17 as a binding partner of CPhar in regulating CPhar downstream factor ATF7 by sequestering C/EBPβ. Conclusions: Our study of this lncRNA CPhar provides new insights into the regulation of exercise-induced cardiac physiological growth, demonstrating the cardioprotective role of CPhar in the heart, as well as expanding our mechanistic understanding of lncRNA function.

Comprehensive Analysis of Key m6A Modification Related Genes and Immune Infiltrates in Human Aortic Dissection

“…DDX17 can regulate smooth muscle cells together with DDX5, through regulating cell growth and division (29). As shown in a study, DDX17, a binding partner of cardiac physiological hypertrophy-associated regulator (CPhar), has the effect of regulating CPhar downstream factor ATF7 (activating transcription factor 7), and CPhar has cardioprotective effect (30). CTGF, also called CCN2, encodes a type of mitogen secreted by vascular endothelial cells (31).…”

Animal exercise studies in cardiovascular research: Current knowledge and optimal design—A position paper of the Committee on Cardiac Rehabilitation, Chinese Medical Doctors’ Association

“…Specific biochemical validation experiments are needed for noncoding RNAs, such as circular RNAs and long non-coding RNAs, that are identified in the hearts of exercised animals. 174 , 175 Next, it is essential to conduct functional experiments in order to determine the roles of these targets in the hearts of exercised animals. Increasing evidence indicates that targets identified in exercised animal hearts have the potential to mitigate cardiac injury in multiple ways.…”

“…In in vitro experiments, primary cultured neonatal rat or mouse cardiomyocytes are essential for determining the functional roles of exercise-responsive molecules in regulating the physiological hypertrophy and the proliferation markers in cardiomyocytes. 119 , 175 The oxygen glucose deprivation/reperfusion-induced apoptosis model can be used to examine whether these molecules have antiapoptosis effects in cultured neonatal cardiomyocytes. 21 , 95 It is noteworthy that cardiomyocytes derived from human embryonic stem cells or human-induced pluripotent stem cells are valuable in confirming these functions in human cardiomyocytes.…”

“… 177 , 178 In in vivo experiments, exercise-associated cardiac phenotypes, such as physiological cardiac hypertrophy (without dysregulation of pathological hypertrophy markers, e.g., atrial natriuretic peptide and brain natriuretic peptide) and increased expression of proliferation markers (e.g., 5-ethynyl-2’-deoxyuridine, Ki-67, phosphohistone H3, and Aurora B) in the myocardium deserve to be validated in exercised animal models such as murine models of swimming exercise by using genetically engineered or modified animals. 92 , 98 , 175 , 179 A deep understanding of the functions and mechanisms of exercise-responsive molecules paves the way for identifying novel myocardial protective or therapeutic targets for CVDs. 8 , 137 , 176 …”

The Relationship among Physical Activity, Intestinal Flora, and Cardiovascular Disease

“…In a study comparing the GM of male rugby players with healthy controls, it was found that the number and diversity of GM of professional rugby players were significantly higher than that of the general population with the same age and BMI [ 14 ]. This could be due to the fact that increasing protein intake through exercise training can promote the diversity of GM [ 15 ].…”