Mitophagy as a regulator of cardiac function in physiological and pathophysiological conditions

7.项目总结 线粒体是一种重要的能量来源，在心脏发育过程中发挥着重要作用 在心力衰竭的发展过程中。累积的证据说明了线粒体质量控制的重要性 在胎儿生命和成人心脏中的心脏功能。此K22应用程序以一项研究提案为中心 为了研究心肌有丝分裂的作用，线粒体质量控制的一个重要方面，在两者中 在职业发展计划范围内的生理和病理生理学条件。这个 该计划旨在促进从博士后培训到独立研究的成功过渡。 这些研究将由我最近描述的监测活体心脏的mt-keima小鼠模型来促进 有丝分裂通量，以及遗传Parkin缺陷(一种阳性有丝分裂调节因子)和USP30的使用 缺陷(负有丝分裂调节)小鼠模型。此外，还将补充详细的心脏 应用小鼠横断主动脉缩窄致心肌病模型进行功能分析。特别是， 我将确定有丝分裂在围产期和心脏线粒体成熟过程中的作用。 病理应激，以及测试一种刺激心脏有丝分裂的特异性USP30抑制剂 延缓心力衰竭进展(目标1)。越来越多的人认识到有丝分裂对于 正常心脏发育和各种疾病中线粒体的可塑性和代谢重编程 条件。我将进一步测试脂肪酸氧化限速酶CPT1a可能 规范有丝分裂。使用心脏特异的CPT1A基因敲除小鼠，我将评估CPT1A是否调节 围产期线粒体代谢转变和成人心脏手术后的有丝分裂反应 压力(目标2)。完成拟议的研究战略将产生对以下方面作用的重要见解 在正常的心血管生理和病理条件下有丝分裂，并将从根本上促进 我们对线粒体代谢和线粒体质量控制之间相互作用的理解 心。这种对有丝分裂素体、USP30和CPT1A在心脏中的作用的理解应该是开放的 利用这些途径实现治疗潜力的可能性。这些研究将在 美国国立卫生研究院的内部计划，在独立的外部阶段完成。 通过这项K22职业发展奖提案，我寻求系统地获得更多 通过详细的职业生涯指导NIH/NHLBI的研究培训和职业发展培训 为补充我目前的技能而设计的发展计划，包括额外的心脏正规培训 生理学和病理生理学。在我的咨询委员会成员的持续支持下，K22 职业发展奖将建立一个培训框架，以启动准备中的研究计划 为了我的独立事业。K22奖项的内部阶段的中心部分将是我的建议 委员会将评估我在拟议的研究和职业发展培训方面的进展情况 在详细的职业发展计划中概述。咨询委员会由校内和 校外成员将提供持续指导。科学培训将支持拟议的具体 旨在通过专业课工作和动手培训相结合的方式来完成拟议的 创新研究战略。重要的是，在资助期间开发的技术和方法 获奖期间不仅将增进我们对心脏有丝分裂的生理作用的了解，而且还将 使拟议的研究战略得以顺利完成。这将为我的第一个 NIH R01和其他独立资金申请。 我还将在这个奖项的内部阶段进行广泛的职业和专业培训，以帮助 在该奖项的校外阶段，预计将面临硕士学位的学术挑战。指导和教学将是 辅以研讨会和讲习班形式的管理和领导能力培训。这个 职业生涯发展培训还包括对初级科学家的指导，参与赠款- 写作研讨会、交流技能的发展、会议上的网络交流、职业咨询和 评估指导，为我向独立的过渡和成为一名 成功的独立调查员。 在美国，心血管疾病是主要的死亡原因，了解其发病机制 调节心脏有丝分裂以保护心脏免受心力衰竭和心肌肥厚的影响可以证明 对公众健康来说是无价的。结构性有丝分裂是维持线粒体的一种动态平衡机制 线粒体的质量和整体功能不仅在心脏，而且在其他组织中也是如此。随着我事业的发展， 我设想我将使用我从研究心脏有丝分裂的功能中获得的洞察力，并应用这一点 了解在其他器官和系统中，特别是在各种情况下吞噬有丝分裂的作用 在大脑中，我已经证明了有丝分裂在神经元退化中的重要作用。总而言之， 拟议的研究将阐明有丝分裂在围产期和成人心脏中的重要性。这些见解 可能为多种心脏疾病的新治疗方法提供基础。此外， 所描述的职业发展计划将显著增强我向学术独立的过渡和机会 继续取得科学上的成功。

7. Project Summary Mitochondria provide an essential source of energy and play an important role during cardiac development and in heart failure progression. Cumulative evidence illustrates the importance of mitochondrial quality control in cardiac function during fetal life and in the adult heart. This K22 application centers on a research proposal to study the role of cardiac mitophagy, an important aspect of mitochondrial quality control, in both physiological and pathophysiological conditions within the context of a career development program. The program is designed to facilitate a successful transition from postdoctoral training to independent research. These studies will be facilitated by my recently described mt-Keima mouse model to monitor in vivo cardiac mitophagic flux, as well as the use of genetic Parkin deficient (a positive mitophagy regulator) and USP30 deficient (a negative mitophagy regulator) mouse models. This will be supplemented by detailed cardiac functional analysis using murine models of transverse aortic constriction induced cardiomyopathy. In particular, I will determine the role of mitophagy in perinatal cardiac mitochondrial maturation and during cardiac pathological stresses, as well as test a specific USP30 inhibitor in stimulating cardiac mitophagy and in attenuating progression of heart failure (Aim 1). It is increasingly recognized that mitophagy is critical for mitochondrial plasticity and metabolic reprogramming during normal heart development and in various disease conditions. I will further test the hypothesis that the rate limiting enzyme in fatty acid oxidation, CPT1a, may regulate mitophagy. Using a cardiac specific CPT1a knockout mouse, I will assess whether CPT1a regulates the mitochondrial perinatal metabolic transition and the adult mitophagic response that occurs following cardiac stress (Aim 2). Completion of the proposed Research Strategy will produce critical insights into the role of mitophagy in normal cardiovascular physiology and pathological conditions, and will fundamentally advance our understanding of the interaction between mitochondrial metabolism and mitochondrial quality control in the heart. This enhanced understanding of the role of mitophagy, USP30 and CPT1a in the heart should open possibilities for harnessing these pathways for therapeutic potential. These studies will be initiated within the NIH intramural program and completed during an extramural, independent phase. Through this K22 Career Development Award proposal, I seek to systematically acquire additional mentored research training and career development training at the NIH/NHLBI through a detailed Career Development Plan designed to complement my current skill set, including additional formal training in cardiac physiology and pathophysiology. With the continued support of members of my Advisory Committee, the K22 Career Development Award will establish a training framework to initiate the research program in preparation for my independent career. A central part of the intramural phase of the K22 award will be my Advisory Committee that will evaluate my progress on the proposed research and career development training as outlined in the detailed Career Development Plan. The advisory committee composed of intramural and extramural members will provide continuous guidance. The scientific training will support the proposed Specific Aims through a combination of specialized course work and hands-on training to complete the proposed innovative Research Strategy. Importantly, the techniques and approaches developed during the funding period of the award will not only advance our understanding of physiological role of cardiac mitophagy, but also allow for the successful completion of the proposed Research Strategy. This will establish the basis of my first NIH R01 and additional independent funding applications. I will also undertake extensive career and professional training in the intramural phase of this award to help master academic challenges anticipated in the extramural phase of the award. Mentoring and teaching will be complemented with training in management and leadership in the form of seminars and workshops. The professional career development training also involves mentoring of junior scientists, participation in grant- writing workshops, development of communication skills, networking at meetings, career counseling and assessment coaching to prepare for my transition to independence and my long term goal of becoming a successful independent investigator. Cardiovascular disease represents the leading cause of death in the USA, understanding the mechanisms regulating cardiac mitophagy that protect the heart from heart failure and cardiac hypertrophy could prove invaluable to public health. Constitutive mitophagy is a homeostatic mechanism for maintaining mitochondrial quality and global mitochondrial function not only in the heart, but also in other tissues. As my career develops, I envision that I will use the insight I have gained from studying the function of cardiac mitophagy and apply this knowledge to investigate the role of mitophagy in a variety of contexts in other organs and systems, particularly in the brain, where I have demonstrated an important role of mitophagy in neuronal degeneration. In summary, the proposed studies will illustrate the importance of mitophagy in the perinatal and adult heart. These insights may provide the basis for novel therapeutic approaches in a wide variety of heart diseases. In addition, the described career development plan will notably enhance my transition to academic independence and chances for continued scientific success.