Mechanisms of Reduced Fatty Acid Oxidation and Cardiac Dysfunction in Sepsis

描述(由申请人提供)：拟议的研究性研究将有助于改善首席调查员(PI)的教育和职业目标。此外，PI将研究脓毒症期间心脏脂肪酸氧化减少的机制。该协会已为拟议研究的头两年(K99阶段)制定了教育计划。该计划包括有关生物信息学工具、拨款申请和负责任的研究行为的课程。此外，他的导师和其他专家将对他的研究技术进行培训，这些专家是他的咨询委员会的成员。PI将进行实验，以研究可以增加心脏脂肪酸氧化的机制，并将为心脏感染性休克的治疗提供治疗方法。在指导阶段结束后，PI将建立一个数据、知识和新技能池，这将增强他成功过渡到独立的资历。在K99阶段，PI计划参加一个关于生物医学信息学的课程，一个关于拨款撰写的课程，另一个关于负责任的研究行为的课程。此外，他还与他的导师和他的顾问/合作者制定了一项培训计划，为他提供如何进行超声心动图、动物模型心力衰竭诱导、染色质免疫沉淀和高通量分析工具和方法的技术知识。这些技术代表着有用的工具，他将能够转移到自己的实验室。除了他将获得的新技术外，拟议中的项目还需要他最近才接触到的领域的知识，例如炎症和类似Krppel的因子生物学。因此，他在他的顾问委员会中包括了一些在这些特定领域具有杰出职业生涯的科学家，他们将提供尽可能高水平的指导。他的所有顾问都受雇于一流的学术部门。他们同意邀请PI在他们的研究所内展示他的工作，以便 他接受了广泛的科学受众的意见，并扩大了他的科学网络。这将促进他向独立研究职位的过渡，并为他申请R01级资金提供坚实的科学基础。该提案的核心问题是：(1)刺激能量产生能否预防内毒素介导的心功能障碍？(2)内毒素如何导致心脏能量学的变化？(3)如何在内毒素治疗的动物中防止粮农组织的减少？为了解决这些问题，国际和平研究所设计了一个由两个分支机构组成的实验计划。第一个阶段为K99阶段，以目前提交的初步数据为基础。这组实验旨在确定PPAR？当PPAR下调时，一种更有效的脂肪酸氧化激活剂，就像脓毒症一样。此外，PI还将研究脓毒症激活的JNK信号通路在减少PPAR？表达方面的作用，PPAR？是一种与脂肪酸氧化密切相关的蛋白质。此外，将建立一种新的具有心肌细胞特异性KLF5缺失的动物模型，并测试其对脓毒症期间心功能障碍的抵抗力。此外，这种新的动物模型将具有高通量分析方法的特点，并可能出现新的靶点。他计划的第二个分支进入了R00阶段。已在初步结果中成功使用并预防败血症的某些干预措施将被探索其在心力衰竭中的治疗潜力。将在K99阶段研究的机制的阐明也将确定新的靶点，并可能为脓毒症的治疗提供新的方法。这些成功干预措施的应用，以及高通量分析将表明的新因素的靶向，将被用于预防脓毒症引起的心功能障碍，以及治疗其他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，在他接受了新技术和心力衰竭模型的培训后，他将更容易过渡到独立的位置，因为他应用预防脓毒症相关性心功能障碍和压力超负荷心力衰竭的方法的能力增强，旨在为这些疾病提供新的治疗方法。总体而言，目前的提议将为PI配备新的知识和有用的工具，以继续在分子心脏病学和应激信号方面的独立职业生涯。他的培训计划旨在促进哥伦比亚大学医学系突出环境中完美的数据生产，以及PI对现代生物医学研究工具的熟悉，他将把这些工具转移到他的独立实验室。对他的K99R00奖申请的批准将导致对新机制的准确定义，这些机制影响心脏脂肪酸氧化和 从而为感染性休克和心力衰竭的治疗提供潜在的新疗法。

DESCRIPTION (provided by applicant): 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.