Role of miR-6236 in Obesity-Associated Adipose Tissue Dysfunction

项目摘要/摘要 这个拟议的为期两年的项目直接源于与我的 目前的K08(DK116668)，并代表了一个新的研究方向，将促进我的进步 通过生成初步数据和出版物实现独立性，以支持R01级应用程序 脂肪组织巨噬细胞(ATM)如何影响脂肪组织动态平衡和机体代谢。 在我获得K08奖的头两年里，我发表了多篇论文，获得了终身教职助理 在宾夕法尼亚大学(宾夕法尼亚大学)担任教授，并在 费城儿童医院(CHOP)。在米切尔·拉扎尔博士和我的K08的持续指导下 咨询委员会，我的独立研究实验室在第一年就大幅增长，现在排名第六 全职研究人员、研究生和博士后。我已经获得了独立的基金会支持 我的实验室，以及NIH的一份附录，研究自动取款机在艾滋病毒相关代谢功能障碍中的作用。我们有 出版了我们第一个完全独立(资深作者)的研究手稿，我被邀请在 多个国家会议。我的实验室现在准备加快我们在ATM机上的研究计划 用于过渡到R01资金。 我的R03提案侧重于了解一种新型的、由ATM分泌的microRNA(miR-6236)如何影响 肥胖背景下的脂肪细胞功能和机体新陈代谢。MIR-6236既是一种新的miRNA，也是一种新的miRNA 最高表达和最高分泌的ATM miRNA。为了促进我们对这种分子的研究，我们有 开发了两种新的转基因小鼠模型，允许整个生物体或组织特异性功能丧失 体内对miR-6236的抑制作用。MiR-6236的全身缺失会导致体重增加和血糖受损 在肥胖背景下的控制。我们还开发了体外模型，初步表明miR- 6236控制脂肪细胞的线粒体呼吸。总而言之，这些数据支持我们的主要假设 MIR-6236通过影响ATM机的功能和相互作用来预防肥胖及其后遗症 组织环境。我们将通过将已建立的miR-6236 loxP小鼠品系与 LysM-Cre大多数菌株，有效地删除了髓系免疫谱系中的miR-6236。这项提案的目标是 是全面表型髓系特异性miR-6236缺陷对脂肪组织的影响 巨噬细胞、脂肪细胞和哺乳动物的代谢。这项工作是我的K08的自然延伸，并将发挥作用 作为我实验室未来研究自动取款机分泌的miRNA的范例。重要的是，这项提案将支持 为该领域的R01级应用程序生成出版物和初步数据。

PROJECT SUMMARY/ABSTRACT This proposed two-year project stems directly from studies and career development activities related to my current K08 (DK116668), and represents a new research direction that will enhance my advancement towards independence through the generation of preliminary data and publications to support an R01-level application on how adipose tissue macrophages (ATMs) influence adipose tissue homeostasis and organismal metabolism. During the first two years of my K08 award I published multiple papers, obtained a tenure-track assistant professorship at the University of Pennsylvania (Penn), and initiated my independent research program at Children’s Hospital of Philadelphia (CHOP). With the continued mentorship of Dr. Mitchell Lazar and my K08 advisory committee, my independent research lab has grown dramatically in its first year and now numbers six full-time research staff, graduate students, and post-docs. I have obtained independent foundation support for my lab, and an NIH supplement to study the role of ATMs in HIV-associated metabolic dysfunction. We have published our first fully-independent (senior author) research manuscripts, and I have been invited to speak at multiple national conferences. My lab is now ready to accelerate our research program in ATMs in preparation for the transition to R01 funding. My R03 proposal focuses on understanding how a novel, ATM-secreted microRNA (miR-6236) influences adipocyte functions and organismal metabolism in the context of obesity. miR-6236 is both a novel miRNA, and the most highly expressed and highly secreted ATM miRNA. To facilitate our studies of this molecule, we have developed two novel transgenic mouse models that allow for whole organism or tissue-specific loss-of-function of miR-6236 in vivo. Whole body deletion of miR-6236 leads to increased weight gain, and impaired glucose control in the context of obesity. We have also developed in vitro models that preliminarily suggest that miR- 6236 controls mitochondrial respiration in adipocytes. Together, these data support our primary hypothesis that miR-6236 protects against obesity and it’s sequela by influencing how ATMs function in, and interact with, their tissue environment. We will test this hypothesis by crossing an established miR-6236 LoxP mouse line to the LysM-Cre most strain, effectively deleting miR-6236 in the myeloid immune lineages. The goal of this proposal is to comprehensively phenotype the effects of myeloid-specific miR-6236 deficiency on the adipose tissue macrophages, adipocytes, and mammalian metabolism. This work is a natural extension of my K08, and will act as a paradigm for future studies of ATM-secreted miRNAs in my lab. Importantly, this proposal will support the generation of publications and preliminary data for an R01-level application in this field.