Dissecting a post-translational modification code in cardiac reprogramming

剖析心脏重编程中的翻译后修饰代码

• 批准号: 10434240
• 负责人: Zhong Wang
• 金额: $ 46.55万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10434240
• 关键词: Acetylation; Animal Model; Automobile Driving; Binding; Cardiac; Cardiac Myocytes; Cell Differentiation process; Cell Fate Control; Cells; Chromatin; Code; Development; Enzymes; Epigenetic Process; Evolution; Fibroblasts; Foundations; GATA4 gene; Gene Expression Regulation; Genetic Transcription; HDAC4 gene; Histone H3; Histones; Human; Investigation; Knowledge; Mediating; Methylation; Molecular; Mus; Myocardial Infarction; Output; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Play; Post-Transcriptional Regulation; Post-Translational Protein Processing; Process; Protein Phosphatase 2A Regulatory Subunit PR53; Proteins; Role; System; Testing; Therapeutic; Translating; Work; cardiac repair; chromatin remodeling; clinical application; clinically relevant; epigenetic regulation; histone modification; in vivo; insight; mouse model; myocyte-specific enhancer-binding-factor 2C; novel; organ growth; organ regeneration; pre-clinical; programs; tandem mass spectrometry; transcription factor

During development and evolution, cell fate changes are guided by cell’s ability to change its transcriptional program with posttranslational modifications (PTMs) on both proteins and chromatin. PTMs of histones have been extensively studied as a coding system to guide alterations in cell plasticity and cell fate, such as differentiation/programming and de-differentiation/reprogramming. The histone PTM coding system includes phosphorylation, acetylation, methylation, and ATP-dependent chromatin remodeling. In parallel, PTMs of free proteins, such as phosphorylation and acetylation of transcription factors, are also essential for organ development and cell differentiation. These PTMs in both histones and free proteins are often catalyzed by the same set of epigenetic enzymes, strongly suggesting a dynamic interplay of PTM coding between histones and epigenetic and transcription factors. However, few systematic studies have been pursued to define the functional output of PTMs as a coding system. Furthermore, the potential synergistic effect of PTMs on both histones and free factors in cell fate changes are poorly explored. Therefore, a deep understanding of the PTM code on both free proteins (ie. key cardiac transcription factors and epigenetic factors) and histones and how they work together will lead to novel insights into cell fate determination and cardiac reprogramming. Our investigation of cardiac reprogramming uncovered a unique phosphorylation code in 14-3-3 binding motifs (PC14-3-3). We hypothesize that activation of a phosphorylation code in 14-3-3 binding motifs triggers a re-arrangement of protein-protein interactome among PC14-3-3 containing reprogramming factors, epigenetic factors, and chromatin, which work synergistically in stimulating cardiac reprogramming. Specific aims: Aim 1: Identify the key factors containing PC14-3-3 essential for cardiac reprogramming. We will test the hypothesis that PC14-3-3 in key reprogramming, epigenetic and histone proteins plays a critical role in reprogramming. These studies will build a knowledge foundation of PC14-3-3 in cardiac reprogramming. Aim 2: Determine PC14-3-3 guided molecular mechanism in stimulating cardiac reprogramming. We will test the hypothesis that PC14-3-3 activation triggers a protein-protein dynamic interplay among PC14-3-3 code containing reprogramming factors (Mef2c and Gata4), epigenetic factors (Hdac4 and Brg1), and histones H3 (H3S10 and H3S28). These studies will establish PC14-3-3 directed epigenetic and chromatin landscape changes in driving cardiac reprogramming. Aim 3: Examine PC14-3-3-guided cardiac reprogramming toward therapeutic applications. We will test the hypothesis that PC14-3-3 stimulates cardiac reprogramming in human cells and clinically relevant animal models. These studies will establish the pre-clinical application of PC14-3-3 code in cardiac reprogramming and build a novel and efficient platform for reprogramming-mediated heart repair.

在发育和进化过程中，细胞命运的变化是由细胞改变其转录的能力引导的。 对组蛋白和染色质进行翻译后修饰 (PTM) 的程序。 作为指导细胞可塑性和细胞命运改变的编码系统已被广泛研究，例如 组蛋白 PTM 编码系统包括分化/编程和去分化/重编程。 磷酸化、乙酰化、甲基化和 ATP 依赖性染色质重塑同时发生。 蛋白质，例如转录因子的磷酸化和乙酰化，对于器官也是至关重要的。 组蛋白和游离蛋白中的这些 PTM 通常由 同一组表观遗传酶，强烈表明组蛋白和组蛋白之间 PTM 编码的动态相互作用 然而，很少有系统研究来定义表观遗传和转录因子。 PTM 作为编码系统的功能输出此外，PTM 对两者的潜在协同效应。 因此，对细胞命运变化中组蛋白和游离因子的深入了解还很少。 游离蛋白（即关键的心脏转录因子和表观遗传因子）和组蛋白的编码以及如何 他们的共同努力将为细胞命运决定和心脏重编程带来新的见解。 我们对心脏重编程的研究发现了 14-3-3 结合中独特的磷酸化代码 我们追踪了 14-3-3 结合基序中磷酸化代码的激活会触发 含有重编程因子、表观遗传的 PC14-3-3 之间蛋白质-蛋白质相互作用组的重排 因子和染色质，它们在刺激心脏重编程中协同作用。 目标 1：确定包含心脏重编程必需的 PC14-3-3 的关键因素。 PC14-3-3 在关键重编程、表观遗传和组蛋白中发挥关键作用的假设 这些研究将为心脏重编程建立 PC14-3-3 的知识基础。 目标 2：确定 PC14-3-3 刺激心脏重编程的分子机制。 将检验 PC14-3-3 激活触发 PC14-3-3 之间蛋白质-蛋白质动态相互作用的假设 包含重编程因子（Mef2c 和 Gata4）、表观遗传因子（Hdac4 和 Brg1）和组蛋白的代码 H3（H3S10 和 H3S28）。这些研究将建立 PC14-3-3 定向的表观遗传和染色质景观。 驱动心脏重编程的变化。 目标 3：检查 PC14-3-3 引导的心脏重编程的治疗应用。 检验 PC14-3-3 刺激人类细胞心脏重编程和临床相关的假设 这些研究将建立 PC14-3-3 代码在心脏中的临床前应用。 重编程并为重编程介导的心脏修复建立一个新颖有效的平台。