Follistatin regulation of energy and lipid metabolism during progression of atherosclerosis

摘要 动脉粥样硬化是大多数心血管疾病（CVD）的根本原因， 对公众健康的威胁。产热脂肪细胞的激活降低胆固醇水平， 动脉粥样硬化的动物模型。血脂异常患者的产热脂肪细胞活性较低， 特征为高血浆甘油三酯（TG）、低血浆高密度脂蛋白胆固醇（HDL-C） 浓度和较低的解偶联蛋白1（UCP 1）表达。这些在老鼠和人类身上的发现 强调了产热脂肪细胞程序诱导剂作为潜在治疗靶点的重要性， 血脂异常和心血管疾病的治疗。我们发表的和初步的数据表明，卵泡抑素（Fst） 不仅在脂肪细胞布朗宁中起重要作用，而且在脂蛋白代谢中起重要作用，以调节TG、总胆固醇、甘油三酯、胆固醇和甘油三酯的水平。 胆固醇（TC）和HDL-C水平。在这个建议中，我们将测试我们的中心假设，即Fst诱导 脂肪布朗宁防止动脉粥样硬化的发展。我们将在这些条件下测试我们的假设。 具体目的：目的1：我们将确定Fst在脂肪布朗宁、脂蛋白代谢和 动脉粥样硬化的发展。我们将使用以下方法确定Fst组织和系统性过表达的影响： 小鼠低密度脂蛋白受体（LDLR）缺陷WT和脂肪特异性转基因（FstAdQTg）小鼠， 和腺相关病毒1（AAV 1）-FST 344分别注射LDLR-/-小鼠，以确定Fst对LDLR-/-小鼠的作用。 i）脂肪布朗宁、血浆脂蛋白、TG和TC水平， 代谢物和iii）饮食诱导的血脂异常和动脉粥样硬化发展过程中的基因表达。效果 将分析Fst在布朗宁、脂蛋白代谢和动脉粥样硬化进展中的瞬时损失。 注射mPCSK 9的Fstfl/fl / CreAdQ小鼠。目的2：我们将确定Fst赋予 促褐变和抗动脉粥样硬化作用。我们将检测是否有精氨酸酶1（Arg 1）/β-肾上腺素能受体 （β-AR）/p38 MAPK/FGF 21信号通路赋予Fst诱导的促褐变和抗动脉粥样硬化作用 三种动物模型我们将利用基因主动脉病变和基因代谢物相关性数据集从 动脉粥样硬化杂交小鼠多样性小组（HMDP）系统遗传学方法鉴定关键Fst调节 基因和代谢物。我们将进行全面的RNA序列分析， 代谢物分析与药理学抑制研究相结合，以鉴定新的基因和代谢物 参与Fst诱导的脂肪布朗宁及其抗动脉粥样硬化作用。 我们的研究将为Fst的促褐变和抗动脉粥样硬化作用提供重要的新见解 并确定参与有利地改变脂蛋白代谢和发展的关键分子靶点， 动脉粥样硬化通过使用Fst蛋白/肽抑制动脉粥样硬化发展是一种有吸引力的治疗方法 大道成功完成该项目后获得的数据为治疗药物设计提供了依据 用于治疗动脉粥样硬化，使我们能够提交极具竞争力的RO 1类资助申请。

Abstract Atherosclerosis is the underlying cause of great majority of cardiovascular diseases (CVD) and poses a great threat to public health. Activation of thermogenic adipocytes reduces cholesterol levels and protects against atherosclerosis in animal models. Lower thermogenic adipocyte activity in dyslipidemic patients has been characterized by high plasma triglycerides (TG), low plasma high density lipoprotein cholesterol (HDL-C) concentration and lower uncoupling protein 1 (UCP1) expression. These findings in mice and humans underscore the importance of inducers of the thermogenic adipocyte program as potential therapeutic targets for the treatment of dyslipidemia and CVD. Our published and preliminary data demonstrate that follistatin (Fst) plays an important role not only in adipocyte browning but also in lipoprotein metabolism to regulate TG, total cholesterol (TC), and HDL-C levels. In this proposal, we will test our central hypothesis that Fst-induced adipose browning protects against the development of atherosclerosis. We will test our hypothesis under these Specific Aims: Aim 1: We will determine the role of Fst in adipose browning, lipoprotein metabolism and development of atherosclerosis. We will determine the effects of Fst tissue and systemic overexpression using mouse low density lipoprotein receptor (LDLR)-deficient WT and adipose-specific transgenic (FstAdQTg ) mice, and adeno-associated virus 1 (AAV1)-FST344 injected LDLR-/- mice respectively to determine the role of Fst on i) adipose browning, plasma lipoprotein, TG, and TC levels, ii) tissue (adipose, aorta, and liver), plasma metabolites and iii) gene expression during diet-induced development of dyslipidemia and atherosclerosis. Effect of transient loss of Fst on browning, lipoprotein metabolism and atherosclerosis progression will be analyzed in mPCSK9 injected Fstfl/fl / CreAdQ mice. Aim 2: We will determine the molecular mechanisms by which Fst confer the pro-browning and anti-atherogenic effects. We will test whether arginase 1 (Arg 1)/beta-adrenergic receptor (β-AR)/ p38MAPK/FGF21 signaling pathway confer Fst-induced pro-browning and antiatherogenic action in the three animal models. We will utilize gene aortic lesion and gene metabolite correlation dataset obtained from system genetics approach of atherosclerosis hybrid mouse diversity panel (HMDP) to identify key Fst-regulated genes and metabolites. We will perform comprehensive RNA-sequence analysis, and tissue and plasma metabolite analysis combined with pharmacological inhibition studies to identify novel genes and metabolites involved during Fst-induced adipose browning and its anti-atherogenic actions. Our proposed studies will provide significant novel insights into the pro-browning and anti-atherogenic role of Fst and identify key molecular targets involved in favorably altering lipoprotein metabolism and the development of atherosclerosis. Inhibition of atherosclerosis development by using Fst protein/peptide is an attractive therapeutic avenue. Data obtained after successful completion of this project provide rationale for therapeutic drug design for the treatment of atherosclerosis and enable us to submit highly competitive RO1-like grant applications.