The impact of sarcomere protein acetylation in heart failure

肌节蛋白乙酰化对心力衰竭的影响

• 批准号: 10077907
• 负责人: Bradley S Ferguson
• 金额: $ 20.82万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10077907
• 关键词: ACTA1 gene; Acetylation; Adult; Affect; Animals; Biology; Calcineurin; Cardiac; Cardiac Myocytes; Cardiac development; Cardiomegaly; Cardiometabolic Disease; Cardiomyopathies; Cardiovascular Diseases; Cardiovascular system; Cellular biology; Centers of Research Excellence; Cessation of life; Chromatin; DNA; DNA Modification Process; DNA Sequence Alteration; Data; Dependovirus; Development; Diagnosis; Disease; Epigenetic Process; Etiology; Functional disorder; Gene Expression; Gene Silencing; Goals; Health; Heart; Heart Hypertrophy; Heart failure; High Fat Diet; Histone Acetylation; Histone Deacetylase; Histones; Human; Hypertrophy; Knowledge; LIM Domain; Laboratories; Left ventricular structure; Link; Lysine; Mass Spectrum Analysis; Mediating; Modeling; Molecular; Mus; Muscle; Muscle Cells; Muscle Contraction; Muscle function; Mutation; Myocardial dysfunction; Myocardium; Myopathy; Nevada; Obesity; Pathologic; Pathology; Pathway Analysis; Patients; Phosphorylation; Physiological; Play; Post-Translational Protein Processing; Prevalence; Process; Protein Acetylation; Protein Binding Domain; Proteins; Regulation; Reporting; Research; Role; Sarcomeres; Serotyping; Signal Pathway; Signal Transduction; Site; Site-Directed Mutagenesis; Skeletal Muscle; Stimulus; Stress; Tail; Testing; Therapeutic; Work; cardiogenesis; cell motility; coronary fibrosis; diet-induced obesity; heart function; histone acetyltransferase; improved; in vivo; innovation; insight; mortality; muscular structure; new therapeutic target; non-histone protein; obese person; response; transcriptome

PROJECT SUMMARY Lysine acetylation has traditionally been studied as an epigenetic modifier of histone tails within chromatin that provides an important mechanism for regulating gene expression. In the heart, histone acetylation acts as a key regulator of cardiac remodeling and function. However, recent reports have shown that non-histone proteins can be acetylated. Importantly, it has been postulated that the acetylome rivals phosphorylation in prevalence as a post-translational modification. The long-term goal of my lab is to dissect the epigenetic and non-epigenetic actions of lysine acetylation in the regulation of heart failure. The objective of this application is to elucidate the role of acetylation of non-histone proteins in the regulation of cardiac hypertrophy and muscle function, with an emphasis on sarcomeric proteins. Preliminary findings from our lab demonstrate that obesity-mediated cardiac remodeling is associated with significant changes in lysine acetylation of proteins within the left ventricle of mice. Mass spectrometry analyses further demonstrated that, of the 3264 lysine-acetylated non-histone proteins identified, 145 were acetylated on 189 unique acetylation sites, 16 of which were significantly impacted by obesity. Ingenuity Pathway Analysis identified the Cardiovascular Disease Network and revealed LIM domain- binding protein 3 (LDB3) and skeletal muscle alpha actin 1 (ACTA1) as proteins that were significantly impacted by obesity. LDB3 and ACTA1 affect muscle structure, integrity, and cellular motility. In addition, LDB3 has been reported to regulate calcineurin-NFAT signaling pathway, which is important in the development of cardiac hypertrophy. Mutations in LDB3 or ACTA1 have been linked to cardiomyopathies, but whether these proteins are acetylated in the heart remains unknown. This proposal will test the central hypothesis that acetylation of sarcomere proteins, specifically LDB3 and ACTA1, regulate cardiac functions through effects on cardiac hypertrophy and muscle contractility. We have developed three specific aims to test this hypothesis. In Aim 1, we seek to delineate a role for sarcomere protein acetylation in cardiac myocyte function. In Aim 2, we elucidate which proteins regulate sarcomere protein acetylation in cardiac myocytes. And in Aim 3, we use adeno- associated virus serotype 9 (AAV9) to determine the physiological significance of LDB3 and ACTA1 acetylation in vivo. Most studies to date have examined lysine acetylation in the regulation of nucleosomal DNA and gene expression. As such, the proposed research is innovative and will add significant insight into the process of sarcomere protein (non-histone) acetylation in the regulation of cardiac biology.

项目摘要 赖氨酸乙酰化传统上被研究为染色质内组蛋白尾部的表观遗传修饰剂， 提供了一种重要的基因表达调控机制。在心脏中，组蛋白乙酰化作为一个关键 心脏重塑和功能的调节剂。然而，最近的报道表明，非组蛋白蛋白可以 被乙酰化。重要的是，据推测，乙酰组竞争对手的磷酸化的流行，作为一个 翻译后修饰我实验室的长期目标是研究表观遗传和非表观遗传 赖氨酸乙酰化在心力衰竭调节中的作用。本申请的目的是阐明 非组蛋白乙酰化在心肌肥大和肌肉功能调节中的作用， 强调肌节蛋白。我们实验室的初步发现表明，肥胖介导的心脏 重塑与小鼠左心室内蛋白质的赖氨酸乙酰化的显著变化有关。 质谱分析进一步表明，在3264个赖氨酸乙酰化的非组蛋白中， 鉴定出145个在189个独特的乙酰化位点上乙酰化，其中16个受到 肥胖伤害途径分析确定了心血管疾病网络，并揭示了LIM结构域- 结合蛋白3（LDB 3）和骨骼肌α肌动蛋白1（ACTA 1）作为蛋白质， 肥胖症。LDB 3和ACTA 1影响肌肉结构，完整性和细胞运动性。此外，LDB 3已 据报道，调节钙调神经磷酸酶-NFAT信号通路，这是重要的心脏的发展， 肥厚LDB 3或ACTA 1的突变与心肌病有关，但这些蛋白质是否 在心脏中的乙酰化程度仍然未知。这一提议将检验中心假设，即乙酰化的 肌节蛋白，特别是LDB 3和ACTA 1，通过对心脏的作用来调节心脏功能。 肥大和肌肉收缩性。我们制定了三个具体目标来检验这一假设。在目标1中， 我们试图描述肌节蛋白乙酰化在心肌细胞功能中的作用。在目标2中，我们阐明 所述蛋白质调节心肌细胞中的肌节蛋白乙酰化。在目标3中，我们使用腺- 相关病毒血清型9（AAV 9），以确定LDB 3和ACTA 1乙酰化的生理意义 in vivo.迄今为止，大多数研究已经检查了赖氨酸乙酰化在核小体DNA和基因调控中的作用。 表情因此，拟议的研究是创新的，将增加对这一过程的重要见解 肌节蛋白（非组蛋白）乙酰化在心脏生物学调节中的作用。