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

摘要 应聘者是一名内分泌学家，在基础科学方面有很强的训练，特别是在脂类领域。 并致力于将这些方法应用于胰岛脂肪酸代谢的研究。这个 候选人的长期职业目标是成为一名独立资助的内科科学家，整合基础 科学研究与临床实践相结合，诊断和治疗代谢性疾病。这个 应聘者的短期职业目标是：1)拓展最新的分子生物学、细胞 生物学、代谢组学和综合生理学技术(包括动物模型)，2)发展强大 假设驱动的研究计划，继续关注新陈代谢机制，以及3)开发 写作技能、资助金和在未来独立资助中变得有竞争力所需的技能 提案。拟议的研究计划、职业发展活动、指导团队和机构 所有这些环境都特别适合帮助申请者实现这些目标。我们的假设是 将脂肪酸激活为长链酰基-辅酶A(LC-COA)的酶，即长链酰基-辅酶A合成酶 (ACSLS)，胰腺SS细胞调节特定的LC-COAs池，这些池在葡萄糖-COAs中发挥作用。 刺激胰岛素分泌(GSIS)和整体胰腺SS细胞功能。在本提案中，PI将测试 一种特定的假设，即特定的酰基-辅酶A合成酶产生LC-COA，并将其导向 增强GSIS的信号通路和另一个特异的酰基辅酶A合成酶通道LC-COAS 转向其他合成和能源生产途径。在目标1中，PI将有选择地击倒 长链酰辅酶A合成酶4(ACSL4)在小鼠胰岛中的表达及其功能 腺病毒RNAi技术和检测基因敲除对GSIS和胰岛脂质介导的影响 发信号。在目标2中，PI将选择性地敲除长链酰辅酶A合成酶5的表达 用腺病毒RNAi技术在小鼠胰岛中表达ACSL5，并检测基因敲除对胰岛的影响 脂肪合成和ss-氧化。这项建议的预期结果是提高对 脂肪酸增强GSIS的机制。为了支持候选人的职业发展，他将 攻读基础质谱学、生物统计分析和研究伦理方面的课程。导师制 团队，其中包括国际公认的、独立资助的具有血脂专业知识的调查人员 新陈代谢(Coleman)和胰岛生物学和新陈代谢(Newgard)将指导Klett博士的研究 和职业发展。研究环境将提供一个高效、合作和协作的环境 实现上述研究和培训目标的氛围。

SUMMARY 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.