Role of acyl-CoA synthetases in mouse pancreatic ??-cell function

描述（由申请人提供）：候选人是一名内分泌学家，在基础科学方面受过良好的培训，特别是在脂质学领域，并致力于将这些方法应用于胰岛脂肪酸代谢的研究。候选人的长期职业目标是成为一名独立资助的医生科学家，将基础科学研究与代谢疾病诊断和治疗的临床实践相结合。候选人的短期职业目标是：1）扩展最新分子生物学，细胞生物学，代谢组学和综合生理学技术（包括动物模型）的技术技能，2）开发强有力的假设驱动的研究计划，继续关注代谢机制，3）开发写作技能，语法和必要的技能，以便在未来的独立资助提案中具有竞争力。拟议的研究计划，职业发展活动，导师团队和机构环境都是唯一适合帮助申请人实现这些目标。我们的假设是，在胰腺β细胞中将脂肪酸活化为长链酰基辅酶A（LC-CoA）的酶，即长链酰基辅酶A合成酶（ACSL），调节在葡萄糖刺激的胰岛素分泌（GSIS）和整体胰腺β细胞功能中发挥功能性作用的特定LC-CoA库。在本提案中，PI将检验特定的假设，即特定酰基辅酶A合成酶产生的LC-CoA被引导至增强GSIS的信号传导途径，另一种特定酰基辅酶A合成酶将LC-CoA引导至其他合成和能量产生途径。在目标1中，PI将使用腺病毒RNAi技术选择性地敲低小鼠胰岛中长链酰基辅酶A合成酶4（ACSL 4）的表达并因此敲低其功能，并检查敲低对GSIS和胰岛脂质介导的信号传导的影响。在目标2中，PI将使用腺病毒RNAi技术选择性地敲低小鼠胰岛中长链酰基辅酶A合成酶5（ACSL 5）的表达，并检查敲低对胰岛脂质合成和β-氧化的影响。这一建议的预期结果是脂肪酸增加GSIS的机制的知识得到了提高。为了支持候选人的职业发展，他将继续学习基础质谱，生物统计分析和研究伦理学。导师团队，其中包括国际公认的，独立资助的研究人员在脂质代谢（科尔曼）和胰岛生物学和代谢（纽加德）的专业知识将指导博士Klett的研究和职业发展。研究环境将提供一个富有成效的，合议和协作的氛围，以追求上述研究和培训目标。 公共卫生相关性：全世界有1.8亿人患有糖尿病，这是一种以血糖升高和胰腺不能分泌胰岛素为特征的慢性疾病。尽管目前的治疗，糖尿病导致显著的发病率，包括心脏病发作，中风，眼病，肾病和肢体丧失。这项研究和后续研究的目标是确定脂肪酸增强胰岛素分泌的机制，这可能导致糖尿病治疗的新策略。

DESCRIPTION (provided by applicant): The candidate is an endocrinologist with strong training in basic science, specifically in the field of lipidology and is committed to applying these methods to the study of fatty acid metabolism in pancreatic islets. The candidate's long-term-career goal is to be an independently funded physician scientist, integrating basic science research with that of clinical practice for the diagnosis and treatment of metabolic disease. The candidate's short-term career goals are to 1) expand technical skills in the latest molecular biology, cell biology, metabolomics, and integrated physiology techniques (including animal models), 2) develop strong hypothesis-driven research program with a continued focus on mechanisms of metabolism, and 3) develop writing skills, grantsmanship and the skills necessary to become competitive in future independent funding proposals. The proposed research plan, career development activities, mentorship team, and institutional environment are all uniquely suited to assist the applicant in achieving these goals. Our hypothesis is that enzymes that activate fatty acids to long-chain acyl-CoAs (LC-CoAs), namely long-chain acyl-CoA synthetases (ACSLs), in pancreatic ss-cells regulate specific pools of LC-CoAs that play a functional role in glucose- stimulated insulin secretion (GSIS) and overall pancreatic ss-cell function. In this proposal, the PI will test the specific hypothesis that a specific acyl-CoA synthetase generates LC-CoAs that are channeled towards a signaling pathway that enhances GSIS and that another specific acyl-CoA synthetase channels LC-CoAs towards other synthetic and energy producing pathways. In Aim 1, the PI will selectively knockdown the expression and therefore function of long-chain acyl-CoA synthetase 4 (ACSL4) in mouse islets using adenoviral RNAi technology and examine the effect of the knockdown on GSIS and islet lipid mediated signaling. In Aim 2, the PI will selectively knockdown the expression of long-chain acyl-CoA synthetase 5 (ACSL5) in mouse islets using adenoviral RNAi technology and examine the effect of the knockdown on islet lipid synthesis and ss-oxidation. The expected outcome of this proposal is an improved knowledge of the mechanisms by which fatty acids augment GSIS. To support the candidate's career development, he will pursue coursework in basic mass spectrometry, biostatistical analysis, and research ethics. The mentorship team, which includes internationally-recognized, independently-funded investigators with expertise in lipid metabolism (Coleman) and pancreatic islet biology and metabolism (Newgard) will guide Dr. Klett's research and career development. The research environment will provide a productive, collegial, and collaborative atmosphere in which to pursue the above research and training goals. PUBLIC HEALTH RELEVANCE: 180 million people worldwide have Diabetes mellitus, a chronic disease characterized by elevated blood glucose and an inability of the pancreas to secrete insulin. Despite current therapies Diabetes mellitus results in significant morbidity, including heart attack, stroke, eye disease, kidney disease, and limb loss. The goal of this research and subsequent research is to determine the mechanisms by which fatty acids enhance insulin secretion that could lead to new strategies for the treatment of diabetes.