Burkholderia-mediated defective killing mechanisms in macrophages from cystic fibrosis (CF) patients

 描述（由申请人提供）：作为一名儿科肺科医生，我的长期职业目标是成为一名医生科学家，他是囊性纤维化（CF）微生物感染免疫控制方面的专家。我的短期教育和研究计划，这个K 08奖的重点是获得科学的独立性，通过详细了解巨噬细胞控制伯克霍尔德菌感染CF。我的住院医师和研究员研究培训使我能够发现小鼠和人类CF模型中巨噬细胞自噬的缺陷，这些缺陷在CF中的伯克霍尔德氏菌毒力中发挥关键作用。新获得的数据表明，伯克霍尔德菌属影响CF巨噬细胞的氧化爆发，但尚不清楚这与CF中的自噬信号传导有何联系。对这些潜在相互交织的途径的追求将产生必要的基础知识，用于开发R 01资助提案，最终目标是未来的新型宿主导向疗法的临床试验，以改善CF中急性和慢性感染的治疗。K 08机制将使我能够将至少75%的精力集中在发展独立的翻译研究事业上。我的职业发展计划将采用一种结构化的方法，具有特定的目标，使我能够过渡到一个独立的研究生涯。我的培训目标包括：1）获得与病原体清除相关的自噬和NADPH氧化酶介导的途径研究的专业知识; 2）操纵巨噬细胞对病原体的信号反应的培训;和3）提高临床前和临床试验开发的转化研究设计技能。我将通过我的研究导师和指导咨询委员会的实践科学培训，相关课程，专业人员公共卫生硕士课程的参与和科学研讨会来实现这些目标。通过这种方法，我将广泛和具体地提高我的研究技能，以充分发展我的独立研究实验室和研究计划，以促进我通过完成K 08奖获得独立资助。该研究计划利用CF中伯克霍尔德氏菌感染的人巨噬细胞模型来研究宿主免疫应答的逃避。我的初步数据表明，伯克霍尔德菌能够抑制CF中已经功能失调的氧化爆发，以避免宿主死亡。这也导致下游自噬体形成减少，进一步减少病原体杀伤。我们假设伯克霍尔德菌在CF中的存活是基于NADPH氧化酶的破坏产生活性氧和自噬体的缺陷组装靶向细胞内细菌杀伤。我们将分析氧化爆发的组装和产生，以及ROS和自噬启动之间的信号相互作用。这项工作的预期结果是，它将促进我们对潜在抗生素的理解- 用于药物递送和/或发现CF中的耐药性伯克霍尔德氏菌感染的独立靶标。这些研究的重要性反映在我们目前缺乏对CF中这些致命感染的金标准治疗。全国儿童医院研究所（RINCH）的微生物发病机制中心和俄亥俄州州立大学的微生物界面生物学中心的环境非常出色，非常适合实现本研究计划和培训目标。我将利用一个熟练的指导团队提供全面的研究指导和培训。Larry Schlesinger博士是巨噬细胞生物学和巨噬细胞-病原体相互作用的世界领导者，拥有长期而杰出的培训记录。他的领导能力与共同导师Amal Amer博士在伯克霍尔德菌发病机制方面的专业知识相结合，包括自噬缺陷的新机制。此外，除了CF特异性细菌发病机制之外，我的导师咨询委员会还在临床、转化和基础CF研究方面拥有更多专业知识。我还拥有在两个最先进的设施中工作的独特优势，并获得临床和基础科学研究核心专家的支持。RINCH的机构支持将使我能够继续将75%的时间投入到拟议的培训和研究中。通过这种跨学科的指导方法，我将大大提高我的科学进步和成功竞争的独立R 01级资金在这个K 08奖的过程中。

 DESCRIPTION (provided by applicant): As a pediatric pulmonologist, my long-term career goal is to become a physician-scientist who is an expert in the immunologic control of microbial infections in cystic fibrosis (CF). My short-term educational and research plans for this K08 award are focused on gaining scientific independence through a detailed understanding of the macrophage control of Burkholderia infections in CF. My resident and fellowship research training has allowed me to discover deficiencies in macrophage autophagy in both murine and human CF models that play a key role in Burkholderia virulence in CF. Newly acquired data suggest that Burkholderia species affect the CF macrophage oxidative burst, but it is unknown how this ties in with autophagy signaling in CF. The pursuit of these potentially intertwined pathways will generate the necessary fundamental knowledge for developing R01 grant proposals with the ultimate goal of future clinical trials of novel host-directed therapies for improving treatment of acute and chronic infections in CF. The K08 mechanism will allow me to focus at least 75% of my efforts on developing an independent translational research career. My career development plan will utilize a structured approach with specific objectives to allow me to transition to an independent research career. My training objectives include: 1) Gaining expertise in the study of autophagy and NADPH oxidase-mediated pathways relating to pathogen clearance; 2) Training in the manipulation of macrophage signaling responses to pathogens; and 3) Enhancing translational research design skills for the development of pre-clinical and clinical trials. I will achieve these objectives through hands-on scientific training ia my research mentors and mentoring advisory committee, relevant coursework, participation in a Master's in Public Health for Professionals program, and scientific seminars. Through this approach I will both broadly and specifically enhance my research skills in order to fully develop my independent research lab and research program to facilitate my achievement of independent funding by the completion of a K08 award. The research plan utilizes a human macrophage model of Burkholderia infection in CF to study evasion of host immune responses. My preliminary data suggests that Burkholderia are able to suppress an already dysfunctional oxidative burst in CF to avoid host killing. This also leads to a downstream reduction in autophagosome formation, further decreasing pathogen killing. We hypothesize that Burkholderia survival in CF is predicated upon both the disruption of the NADPH oxidase for generation of reactive oxygen species and the deficient assembly of autophagosomes for targeted intracellular bacterial killing. We will analyze both the assembly and generation of the oxidative burst, as well as signaling interactions between ROS and autophagy initiation. The anticipated outcome of this work is that it will advance our understanding of potential antibiotic- independent targets for drug delivery and/or discovery for antibiotic-resistant Burkholderia infections in CF. The significance of these studies is reflected in our current lack of a gold standard treatment for these deadly infections in CF. The environments of the Center for Microbial Pathogenesis at the Research Institute at Nationwide Children's Hospital (RINCH) and the Center for Microbial Interface Biology at The Ohio State University are outstanding and ideally suited for the achievement of this research plan and training objectives. I will utilize a skilled mentoring team to provide comprehensive research guidance and training. Dr. Larry Schlesinger is a world leader in macrophage biology and macrophage-pathogen interactions with a long and distinguished training record. His leadership is coupled with co-mentor Dr. Amal Amer's expertise in Burkholderia pathogenesis, including novel mechanisms of autophagy deficiency. Additionally, my mentor advisory committee features further expertise in clinical, translational, and basic CF research in addition to CF-specific bacterial pathogenesis. I also have the unique benefit of working in two state-of-the-art facilities and receiving support from experts in both clinical and basic science research cores. Institutional support from the RINCH will allow me to continue to devote 75% of my time towards the proposed training and research. Through this interdisciplinary mentoring approach I will greatly enhance my scientific progression and successful competition for independent R01 level funding during the course of this K08 award.