Mechanisms of Reduced Fatty Acid Oxidation and Cardiac Dysfunction in Sepsis

摘要 拟议的研究性研究将有助于改善校长的教育和职业目标。 调查员(PI)。此外，PI将研究心脏脂肪酸减少的机制。 脓毒症时的氧化作用。该协会已制定了首两年(K99阶段)的教育计划。 建议的研究。该计划包括关于生物信息学工具的课程，拨款申请和负责任的行为 研究。此外，PI将由他的导师和其他专家进行研究技术培训，这些专家是 他的顾问委员会的成员。PI将进行实验，以研究能够 增加心脏脂肪酸氧化，并将为心脏败血症的治疗提供建议 令人震惊。在指导阶段结束后，PI将建立一个数据、知识和新的 这些技能将增强他成功过渡到独立的资历。 在K99阶段，PI计划参加一门关于生物医学信息学的课程，一门关于补助金撰写的课程 另一个是负责任的研究行为。此外，他还与他的导师制定了培训计划 和他的顾问/合作者，为他提供如何表演的技术知识 超声心动图，动物模型心力衰竭的诱导，染色质免疫沉淀和高密度脂蛋白 吞吐量分析工具和方法。这些技术代表着有用的工具，他将能够转移 送到他自己的实验室。除了他将获得的新技术外，拟议的项目还需要了解 他最近才被与之联系在一起的领域，如炎症和Kr�pPEL样因子生物学。 因此，他在他的顾问委员会中包括了一些在这些领域有杰出成就的科学家。 将提供最高级别指导的特定领域。他所有的顾问都受雇于TOP- 攻克学术院系。他们已经同意邀请PI在他们的研究所内展示他的工作，所以 他接受了广泛的科学受众的意见，并扩大了他的科学网络。这将促进他的 过渡到独立研究职位，并为他提供坚实的科学基础，从 申请R01级资助。 该提案的核心问题是：(1)刺激能量产生能否阻止内毒素介导的心脏 功能障碍？(2)内毒素如何导致心脏能量学变化？(3)粮农组织如何减少 在内毒素治疗的动物中预防？为了解决这些问题，国际和平研究所设计了一项实验计划 有两个分支。第一个阶段进入K99阶段，基于当前的初步数据 呈件。这组实验旨在确定使PPAR�成为更有效的激活剂的机制 当PPAR�下调时，脂肪酸氧化的影响，就像在脓毒症中发生的那样。此外，公安局将调查 脓毒症激活的jnk信号通路在降低PPAR�、a的表达中的作用 与脂肪酸氧化密切相关的蛋白质。此外，一种新的动物模型 将产生心肌细胞特异性的KLF5缺失，并测试其对心功能障碍的抵抗力 在脓毒症期间。此外，这种新的动物模型将具有高通量分析方法的特点 而且可能会出现新的目标。他计划的第二个分支进入了R00阶段。某些干预措施 已经在初步结果中成功应用并预防脓毒症的药物将被探索用于其 心力衰竭的治疗潜力。将在K99中研究的机制的阐明 阶段还将确定新的靶点，并可能为脓毒症的治疗提出新的方法。应用 这些成功的干预措施，以及高通量分析正在进行的新因素的目标 表明将用于预防脓毒症所致的心功能不全，以及用于治疗其他疾病 伴有脂肪酸氧化受损的心功能障碍，如压力超负荷心力衰竭。因此，在 他在新技术和新一代心力衰竭模型方面的训练，他向独立职位的过渡 将被促进，因为他应用预防败血症相关心脏疾病的方法的能力增加了 功能障碍和压力超负荷心力衰竭，旨在为这些疾病提供新的治疗方法。 总体而言，目前的提议将使PI具备新的知识和有用的工具，以便继续开展工作 在分子心脏病学和压力信号方面独立工作。他的训练计划被设计成 在联合国医学部突出的环境中促进数据的无懈可击的产生 哥伦比亚大学，以及私家侦探对现代生物医学研究工具的熟悉，他将 转到他的独立实验室。他的K99R00奖申请获得批准将导致 精确定义影响心脏脂肪酸氧化的新机制，从而提供 治疗感染性休克和心力衰竭的潜在新疗法。

SUMMARY The proposed research study will facilitate the improvement of the education and career goals of the principal investigator (PI). Moreover, the PI will investigate the mechanisms that underlie reduced cardiac fatty acid oxidation during sepsis. The PI has set up an educative plan for the first two years (K99 phase) of the proposed study. This plan includes courses about bioinformatics tools, grant writing and responsible conduct of research. In addition, the PI will be trained in research techniques by his mentor and other experts, who are members of his advisory committee. The PI will perform experiments to investigate mechanisms that can increase cardiac fatty acid oxidation and will suggest therapeutic approaches for the treatment of cardiac septic shock. Upon conclusion of the mentored phase the PI will have established a pool of data, knowledge and new skills that will enhance his credentials for successful transition to independence. During the K99 phase the PI has planned to attend one course on Biomedical Informatics, one on grant writing and another in the responsible conduct of research. In addition, he has set up a training plan with his mentor and his advisors/collaborators that will provide him with technical knowledge on how to perform echocardiography, induction of heart failure in animal models, chromatin immunoprecipitation and high throughput analysis tools and methods. These techniques represent useful tools that he will be able to transfer to his own lab. Besides the new techniques that he will acquire, the proposed project requires knowledge in areas that he has only recently been associated with, such as inflammation and Kr�ppel-like factor biology. Therefore, he has included in his advisory board a number of scientists with outstanding careers in these particular fields that will provide guidance of the highest possible level. All his advisors are employed by top- notch academic departments. They have agreed to invite the PI to present his work within their institutes, so that he receives input by a broad scientific audience and expands his scientific network. This will promote his transition to an independent research position and provide him a robust scientific foundation from which to apply for R01-level funding. The proposal core questions are: (1) Can stimulation of energy production prevent LPS-mediated cardiac dysfunction? (2) How does LPS lead to changes in cardiac energetics? (3) How can the reduction in FAO be prevented in LPS treated animals? To address these questions the PI has designed an experimental plan with two branches. The first falls into the K99 phase and is based on preliminary data of the current submission. This set of experiments aims to identify the mechanism that makes PPAR� a more potent activator of fatty acid oxidation when PPAR� is downregulated, like it happens in sepsis. Also, the PI will investigate the role of the JNK signaling pathway, which is activated by sepsis, in the reduction of the expression of PPAR�, a protein that has been strongly associated with fatty acid oxidation. Furthermore, a new animal model with cardiomyocyte specific deletion of KLF5 will be generated and tested for resistance to cardiac dysfunction during sepsis. Besides, this novel animal model will be characterized with high throughput analysis methods and new targets may come up. The second branch of his plan falls into the R00 phase. Certain interventions that have been used successfully in the preliminary results and prevent sepsis will be explored for their therapeutic potential in heart failure. The elucidation of the mechanisms that will be investigated in the K99 phase will also identify new targets and may suggest novel approaches for the treatment of sepsis. Application of these successful interventions, as well as targeting of new factors that the high throughput analysis is going to indicate will be used to prevent sepsis-mediated cardiac dysfunction, as well as to treat other conditions of cardiac dysfunction with impaired fatty acid oxidation, such as pressure overload heart failure. Therefore, after his training in new technologies and generation of heart failure models, his transition to independent position will be facilitated due to his increased capacity to apply methods that prevent sepsis-associated cardiac dysfunction and pressure overload heart failure, aiming to provide novel treatments for these diseases. Overall, the current proposal will equip the PI with novel knowledge and useful tools to continue for independent career in molecular cardiology and stress signaling. His training plan has been designed to facilitate flawless production of data in the prominent environment of the Department of Medicine of the Columbia University, as well as the PI's acquaintance with modern tools of biomedical research that he will transfer to his independent laboratory. Approval of his application for the K99R00 award will result in the precise definition of novel mechanisms, which affect cardiac fatty acid oxidation and will thereby provide potential new therapeutics for the treatment of septic shock and heart failure.