Role of KLF15 in Circadian Regulation of Cardiac Ischemia

KLF15 在心脏缺血昼夜节律调节中的作用

• 批准号: 9032864
• 负责人: Lilei Zhang
• 金额: $ 16.42万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-15 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9032864
• 关键词: Accounting; Advisory Committees; Affect; Animals; Area; Award; Behavioral; Biology; Blood flow; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Catabolism; Cell model; Cells; ChIP-seq; Chronic; Circadian Rhythms; Clinical; Data; Development; Ectopic Expression; Event; Exhibits; Feedback; Foundations; Funding; Future; Gene Expression Regulation; Gene Targeting; Genes; Genetic Transcription; Genetically Engineered Mouse; Grant; Heart; Heart Diseases; Homeostasis; Hour; Human; Human Genetics; In Vitro; Incidence; Individual; Infarction; Injury; Knockout Mice; Kruppel-like transcription factors; Laboratories; Lead; Link; Liver; Mediating; Medical Genetics; Mentors; Metabolic; Metabolism; Methods; Modeling; Molecular; Molecular Biology; Morbidity - disease rate; Mus; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardium; Oxidation-Reduction; Oxidative Stress; Patients; Periodicity; Peripheral; Phase; Physicians; Physiological; Play; Predisposition; Primary Cell Cultures; Process; Production; Proteins; Reactive Oxygen Species; Regulation; Reperfusion Injury; Reperfusion Therapy; Research; Research Institute; Research Personnel; Research Project Grants; Resting Phase; Rodent; Role; Sampling; Scientific Advances and Accomplishments; Scientist; Severities; Signal Transduction; Skeletal Muscle; Technology; Testing; Therapeutic Agents; Time; Tissues; Training; Transcript; United States National Institutes of Health; Universities; Zinc Fingers; base; cardiac repair; career; cell type; circadian pacemaker; collaborative environment; experience; gene repair; genomic tools; heart function; heart metabolism; high risk; in vivo; in vivo Model; induced pluripotent stem cell; injury and repair; interest; ischemic cardiomyopathy; lipid metabolism; loss of function; metabolic profile; mortality; mouse model; new therapeutic target; next generation sequencing; novel; novel therapeutics; prevent; programs; public health relevance; repaired; response; restoration; sudden cardiac death; therapy development; transcription factor; transcriptome; transcriptomics

 DESCRIPTION (provided by applicant): Cardiovascular diseases are the leading cause of morbidity and mortality in the U.S. It has long been documented that the incidence and severity of myocardial infarction has a diurnal variation, with the peak in the early morning. The molecular basis, however, is poorly understood. Also, state-of-the-art flow restoring therapies cannot avoid the ischemic reperfusion (I/R) injury either at the time of heart attack or during revascularization. Kruppel-like zinc finger transcription factors play important roles in multiple aspects of cardiovascular biology. We have characterized the role of Kruppel-like factor 15 (KLF15) in regulating cellular metabolism in the liver and the skeletal muscle. Nascent observations by the applicant suggest a critical role of KLF15 in coordinating catabolism and reactive oxygen species (ROS) clearance in the heart. KLF15 deficient myocardium/cardiomyocytes exhibits increased accumulation of ROS and exaggerated susceptibility to I/R injury. Using next generation sequencing, we discovered that KLF15 governs 75% of the oscillating transcripts in the heart and organize them into two temporal functional clusters, one enriched for catabolic genes coincides with the active phase, another enriched for remodeling and repair genes coincides with the resting phase. Loss of function of KLF15 in the murine heart leads to aberrant oscillation. Finally, KLF15 is significantly reduced in human heart samples with ischemic cardiomyopathy. Using a combination of murine models and molecular/cellular approaches, Dr. Zhang will study the molecular mechanisms of cardiac KLF15 in the dynamic process circadian regulation of fuel utilization and redox homeostasis. In addition, Dr. Zhang will use novel induced pluripotent stem cell derived cardiomyocyte (iPS-CM) technology to extend the studies to human cells. With the support provided by the K08 award, Dr. Zhang's project will lead to a better understanding of the role of KLF15 on the circadian variability of the susceptibility to I/R injury, reveal the molecular mechanism of such regulation and may provide a foundation for the development of novel therapies to limit myocardial injury. The specific aims of the proposal include: 1. To determine the effect of KLF15 deficiency on cardiac I/R injury: We will use cardiac-specific KLF15 KO mice to assess the cardiac I/R injury, lipid metabolism, ROS and transcriptome in a circadian fashion. 2. To identify and validate key targets of KLF15 in the cardiac circadian gene regulation: We will ChIP-seq to identify direct KLF15 targets and validate them in murine models both in vitro and in vivo. 3. To evaluate the function of KLF15 and its key targets in hiPS-CM: We will assess key findings in murine models in human iPS-CM. The applicant, Dr. Lilei Zhang, a physician-scientist with special interest in cardiogenetics, outlines a 5-year mentored career plan that will build upon her experience in human genetics, molecular biology and clinical medical genetics. Under the guidance of Dr. Mukesh Jain, a recognized leader in transcriptional and cardiovascular biology, Dr. Zhang will use tissue specific murine model combined with genomic tools to study the role of KLF15 in circadian regulation of cardiac metabolism and reduction-oxidation (redox) homeostasis. With assistance from Dr. Anthony Wynshaw-Boris, she will also use (iPS-CM) technology to extend the project to the relevant cell types in human. Dr. Zhang will be mentored by an Advisory Committee composed of highly successful, NIH-funded physician scientists at Case Western Reserve University (CWRU) with expertise in her area of research. This plan is ideally carried out in the Cardiovascular Research Institute (CVRI) at Case Western Reserve University with its distinguished record for training physician-scientists in a rich and collaborative environment. The mentored K08 award will directly advance her scientific development by protecting her effort toward her research project and help her transition into an independent investigator with her own laboratory and R01 funding.

 描述（由适用提供）：心血管疾病是美国发病率和死亡率的主要原因，长期以来一直记录在心肌梗死的事件和严重程度有差异性变化，并且在清晨的峰值。但是，分子基础知之甚少。同样，在心脏病发作或血运重建期间，最先进的恢复疗法无法避免缺血性再灌注（I/R）损伤。 Kruppel样锌指转录因子在心血管生物学的多个方面起着重要作用。我们已经表征了Kruppel样因子15（KLF15）在控制肝脏和骨骼肌中细胞代谢中的作用。适用的新生观察表明，KLF15在心脏中的配位分解代谢和活性氧（ROS）清除率中的关键作用。 KLF15缺乏的心肌/心肌细胞表现出ROS的积累增加，并且夸大了I/R损伤的敏感性。使用下一代测序，我们发现KLF15控制着心脏中的75％的振荡转录本，并将它们组织成两个临时功能簇，一个富含分解代谢基因与活性相一致，另一个富含重塑和修复基因与休息阶段相结合。 KLF15在鼠心脏中的功能丧失会导致异常振荡。最后，KLF15显着降低 人心脏样本患有缺血性心肌病。张博士利用鼠模型和分子/细胞方法的组合，将研究心脏KLF15的分子机制在动态过程中，昼夜节律调节燃料利用率和氧化还原稳态。此外，张博士将使用新型诱导的多能干细胞衍生的心肌细胞（IPS-CM）技术将研究扩展到人类细胞。在K08奖励提供的支持下，张博士的项目将更好地理解KLF15对I/R损伤易感性的昼夜节律可变性的作用，揭示了这种调节的分子机制，并可能为开发新疗法的基础提供了限制心肌损伤的基础。该提案的具体目的包括：1。确定KLF15缺乏对心脏I/R损伤的影响：我们将使用心脏特异性的KLF15 KO小鼠评估心脏I/R损伤，脂质代谢，ROS和转录组的昼夜节律方式。 2。在心脏昼夜节律调节中识别和验证KLF15的关键靶标：我们将chip-seq识别直接的KLF15靶标，并在体外和体内在鼠模型中验证它们。 3。为了评估KLF15及其在HIPS-CM中的关键目标的功能：我们将评估人IPS-CM中鼠模型中的关键发现。申请人Lilei Zhang博士是一位对心脏遗传学特别感兴趣的身体科学家Lilei Zhang博士，概述了一项为期5年的职业计划，该计划将基于她在人类遗传学，分子生物学和临床医学遗传学方面的经验。在转录和心血管生物学领域公认的领导者Mukesh Jain博士的指导下，张博士将使用组织特定的鼠模型与基因组工具相结合来研究KLF15在昼夜节律调节心脏代谢和还原氧化（Redox）稳态中的作用。在Anthony Wynshaw-Boris博士的协助下，她还将使用（IPS-CM）技术将项目扩展到人类相关的细胞类型。张博士将由一个咨询委员会（Case Western Reserve University（CWRU），由NIH资助的非常成功，资助的物理科学家组成，并在其研究领域具有专业知识。理想情况下，该计划是在Case Western Reserve University的心血管研究学院（CVRI）中进行的，其杰出的记录是在丰富且协作的环境中培训物理科学家。这 指导的K08奖将通过保护她在研究项目上的努力并帮助她通过自己的实验室和R01资金过渡到独立调查员，直接推动她的科学发展。