Defining the Role of lnc-TECRL in Maladaptive Cardiac Remodeling

摘要 该提案概述了一项为期4年的综合培训计划，以发展Veli K.Topkara，MD，MSc，And 哥伦比亚大学医学助理教授，成为一名独立的翻译研究员。Dr。 Topkara是一位高级心力衰竭(HF)心脏病专家，他的长期目标是了解基本机制 负责适应性不良的心脏重塑，重点是长的非编码RNA(LncRNAs)，依次为 以改善心力衰竭患者的预后。职业发展计划和导师结构 旨在填补Topkara博士在教育和经验方面的具体差距 培训，确保他成熟为一名独立的内科科学家。他的短期职业目标包括：1) 培养设计和实施机械性翻译研究的技能，重点是lncRNA 生物学，2)获得诱导多能干细胞来源的心肌细胞(iPS-CMS)的专业知识 和基因编辑技术，3)发展lncRNA的计算和生物信息学分析经验， 4)掌握体内lncRNA敲除策略的开发和应用技能，以及5)开发Grant- 写作技巧。导师团队代表了一个由多个学科的个人组成的小组，他们专门被选为 完成PI特定的科学和职业发展目标。在机构层面，哥伦比亚大学提供了一个 令人难以置信的丰富和支持性的智力和协作研究环境，提供充足的资源 托普卡拉博士成功地完成了他的研究和职业发展目标。通过RNA Topkara博士对失败的人类心脏样本进行了测序，确定了81个候选的显著 心衰时调节失调。基于(I)基因-组织表达(GTEx)数据集中的心脏组织富集物，(Ii) 在左心室辅助装置支持下的可逆性，(Iii)在小鼠中的轨迹保守，(Iv)在 心肌细胞肥大；选择LNC-TECRL(LIKE)进行结构分析 和功能表征。根据初步数据，这一建议的主要假设是lnc- TECRL通过与染色质修饰蛋白相互作用介导病理性心肌肥厚 肥大基因程序的激活。具体目标是：1)确定lnc通过哪些机制- TECRL介导体外培养的心肌细胞肥大2)研究LNC-TECRL的治疗潜力 预防体内不良适应性心脏重塑。为了实现这些目标，PI将利用成年人类 心肌细胞，人iPS-CMS，成熟的工程化人类心脏组织，以及将 建立lnc-TECRL在心脏重构中的功能和治疗相关性。要了解机制 通过它介导心肌肥厚，最新的技术包括RNA下拉，RNA-IP， 将采用芯片、结构域缺失映射、chirp-seq和体内GapmeR基因敲除。完成 有望为研究适应不良的转录调控提供新的见解 心脏重塑和发现新的方法，可以转化为治疗心力衰竭患者的好处。 。

Abstract This proposal outlines a comprehensive 4-year training program to develop Veli K. Topkara, MD, MSc, an Assistant Professor of Medicine at Columbia University, into an independent translational investigator. Dr. Topkara is an advanced heart failure (HF) cardiologist whose long-term goal is to understand basic mechanisms responsible for maladaptive cardiac remodeling with an emphasis on long non-coding RNAs (lncRNAs), in order to improve outcomes of patients with heart failure. The career development plan and mentorship structure proposed in this application are designed to fill specific educational and experiential gaps in Dr. Topkara’s training, ensuring his maturation into an independent physician-scientist. His short-term career goals include: 1) to develop skills in design and conduct of mechanistic translational research with a specific focus on lncRNA biology, 2) to acquire expertise in generation of induced-pluripotent stem cell derived cardiomyocytes (iPS-CMs) and gene editing technology, 3) to develop experience in computational and bioinformatics analysis of lncRNAs, 4) acquire skills in development and application of in vivo lncRNA knockdown strategies, and 5) to develop grant- writing skills. The mentorship team represents a multidisciplinary group of individuals specifically chosen to achieve PI's particular scientific and career development goals. At an institutional level, Columbia provides an incredibly rich and supportive intellectual and collaborative research environment with ample resources available for Dr. Topkara to successfully accomplish his research and career development goals. Through RNA sequencing of failing human heart samples, Dr. Topkara identified 81 candidate lncRNAs that are significantly dysregulated in HF. Based on (i) cardiac tissue enrichment in Genotype-Tissue Expression (GTEx) dataset, (ii) reversibility with left ventricular assist device support, (iii) locus conservation in mouse, (iv) modulation during cardiomyocyte hypertrophy; lnc-TECRL (lnc-Trans-2,3-Enoyl-CoA Reductase Like) was selected for structural and functional characterization. Based on preliminary data, the main hypothesis of this proposal is that lnc- TECRL mediates pathological cardiac hypertrophy through its interaction with chromatin modifying proteins and activation of the hypertrophic gene program. The specific aims are to: 1) Determine mechanisms by which lnc- TECRL mediates cardiomyocyte hypertrophy in vitro and 2) Investigate the therapeutic potential of lnc-TECRL in prevention of maladaptive cardiac remodeling in vivo. To achieve these goals, the PI will utilize adult human cardiomyocytes, human iPS-CMs, mature engineered human cardiac tissues, and mouse models which will establish functional and therapeutic relevance of lnc-TECRL in cardiac remodeling. To understand mechanisms by which lnc-TECRL mediates cardiac hypertrophy, state-of-art techniques including RNA pull-down, RNA-IP, CHIP, domain mapping by deletion, ChiRP-seq, and in vivo GapmeR knockdown will be employed. Completion of the proposed work is expected to provide novel insights into the transcriptional regulation of maladaptive cardiac remodeling and uncover new approaches that could be translated for therapeutic benefit in HF patients. .