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(申请人提供)：心血管疾病是美国发病率和死亡率的主要原因。长期以来，有证据表明，心肌梗死的发病率和严重程度有每日变化，高峰在清晨。然而，其分子基础却鲜为人知。此外，最先进的血流恢复疗法无论是在心脏病发作时还是在血运重建期间都无法避免缺血再灌注(I/R)损伤。Kruppel类锌指转录因子在心血管生物学的多个方面发挥着重要作用。我们已经确定了Kruppel-like factor15(KLF15)在调节肝脏和骨骼肌细胞代谢中的作用。申请人的初步观察表明，KLF15在协调心脏分解代谢和清除活性氧(ROS)方面发挥着关键作用。KLF15缺乏的心肌/心肌细胞表现出ROS积聚增加，对I/R损伤的易感性增加。利用下一代测序，我们发现KLF15控制着心脏中75%的振荡转录本，并将它们组织成两个时间功能簇，一个富含分解代谢基因的时间段与活动期一致，另一个富含重塑和修复基因的时间段与静止期一致。KLF15在小鼠心脏中的功能丧失会导致异常振荡。最后，KLF15在 患有缺血性心肌病的人心脏样本。张博士将结合小鼠模型和分子/细胞方法，研究心脏KLF15在燃料利用和氧化还原动态平衡的昼夜节律调节过程中的分子机制。此外，张博士将使用新的诱导多能干细胞来源的心肌细胞(iPS-CM)技术将研究扩展到人类细胞。在K08奖项的支持下，张博士的项目将使人们更好地理解KLF15在I/R损伤易感性的昼夜变化中的作用，揭示这种调节的分子机制，并可能为限制心肌损伤的新疗法的开发提供基础。1.为确定KLF15缺乏对心脏I/R损伤的影响：我们将使用心脏特异的KLF15 KO小鼠，以昼夜节律性的方式评估心脏I/R损伤、脂质代谢、ROS和转录组。2.确定和验证KLF15在心脏昼夜节律基因调控中的关键靶点：我们将利用芯片序列技术来确定KLF15的直接靶点，并在体外和体内模型中对其进行验证。3.评估KLF15在HIPS-CM中的功能及其关键靶点：我们将评估在人iPS-CM小鼠模型中的关键发现。申请者张丽蕾博士是一位对心脏遗传学特别感兴趣的内科科学家，她概述了一份为期5年的指导职业计划，该计划将以她在人类遗传学、分子生物学和临床医学遗传学方面的经验为基础。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异性小鼠模型结合基因组工具来研究KLF15在心脏代谢和还原-氧化(Redox)动态平衡的昼夜调节中的作用。在Anthony Wynshaw-Boris博士的帮助下，她还将使用(iPS-CM)技术将该项目扩展到人类相关细胞类型。张博士将接受一个咨询委员会的指导，该委员会由美国国立卫生研究院资助的凯斯西储大学(CWRU)非常成功的内科科学家组成，他们在她的研究领域拥有专业知识。该计划理想地在凯斯西储大学心血管研究所(CVRI)实施，该研究所在丰富和协作的环境中培训内科科学家方面有着卓越的记录。这个 导师K08奖将直接推动她的科学发展，保护她对研究项目的努力，并帮助她转变为拥有自己的实验室和R01资金的独立研究员。