Mechanisms of Reduced Fatty Acid Oxidation and Cardiac Dysfunction in Sepsis

描述（由申请人提供）：拟议的研究将有助于改善主要研究者（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。PI已为拟议研究的前两年（K99阶段）制定了教育计划。该计划包括有关生物信息学工具，资助写作和负责任的研究行为的课程。此外，PI将接受其导师和其他专家的研究技术培训，这些专家是其咨询委员会的成员。PI将进行实验，以研究可增加心脏脂肪酸氧化的机制，并将提出治疗心脏感染性休克的治疗方法。在指导阶段结束时，PI将建立一个数据库，知识和新技能，这将提高他成功过渡到独立的资格。在K99阶段，PI计划参加一门关于生物医学信息学的课程，一门关于赠款写作的课程，另一门关于负责任的研究行为的课程。此外，他还与他的导师和顾问/合作者一起制定了培训计划，为他提供如何进行超声心动图，在动物模型中诱导心力衰竭，染色质免疫沉淀和高通量分析工具和方法的技术知识。这些技术代表了有用的工具，他将能够转移到他自己的实验室。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，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特定领域具有杰出职业生涯的科学家，这些科学家将提供尽可能高水平的指导。他所有的顾问都受雇于一流的学术部门。他们同意邀请PI在他们的研究所内介绍他的工作，以便 他接受了广泛的科学观众的投入，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R 01级资助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LPS介导的心功能不全？(2)LPS如何导致心脏能量学的变化？(3)在LPS处理的动物中如何防止FAO的减少？为了解决这些问题，PI设计了一个有两个分支的实验计划。第一份福尔斯报告属于K99阶段，以本次划界案的初步数据为基础。这组实验的目的是确定机制，使过氧化物酶体增殖物激活受体？当过氧化物酶体增殖物激活物受体被下调时，它是一种更有效的脂肪酸氧化激活剂，就像败血症一样。此外，PI将研究JNK信号通路的作用，这是由脓毒症激活，在减少表达的过氧化物酶体增殖物激活受体，一种蛋白质，已与脂肪酸氧化密切相关。此外，将产生具有心肌细胞特异性KLF 5缺失的新动物模型，并测试其在脓毒症期间对心功能障碍的抵抗力。此外，这种新型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。他的计划的第二个分支福尔斯落入R 00阶段。在初步结果中已成功使用并预防脓毒症的某些干预措施将探索其在心力衰竭中的治疗潜力。在K99阶段研究的机制的阐明也将确定新的目标，并可能提出治疗脓毒症的新方法。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，将用于预防脓毒症介导的心功能不全，以及治疗其他脂肪酸氧化受损的心功能不全疾病，如压力超负荷心力衰竭。因此，在他接受新技术和心力衰竭模型的培训后，由于他应用预防脓毒症相关的心功能不全和压力超负荷心力衰竭的方法的能力增加，他向独立职位的过渡将得到促进，旨在为这些疾病提供新的治疗方法。总体而言，目前的提案将为PI提供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在哥伦比亚大学医学系的突出环境中完美地生产数据，以及PI对他将转移到他的独立实验室的现代生物医学研究工具的熟悉。批准他的K99 R 00奖申请将导致对新机制的精确定义，这些机制影响心脏脂肪酸氧化， 从而将为治疗败血性休克和心力衰竭提供潜在的新疗法。

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.