Omega-3 PUFA suppress adipochemokines in human obesity

描述（由申请人提供）：肥胖及其并发症代表了一个主要的公共卫生;进一步了解潜在的病理生理学将有助于预防和治疗方式的发展。作为一名儿科内分泌学家，私家侦探。Shah博士致力于研究肥胖相关代谢失调的早期机制过程，希望探索新的预防和治疗方案。她的最终职业目标是成为内分泌学领域的独立翻译研究人员，专注于儿童和成人年龄范围内的肥胖和脂肪组织生物学。通过她目前的工作与她的导师和在硕士转化研究计划的培训，她已经开发了临床和转化研究的基本技能和经验。这将使她能够执行本申请中提出的工作，该申请将人体临床试验与人体组织和细胞的研究相结合，以探索肥胖相关脂肪炎症的新途径。为了获得更多的专业知识和知识，她过渡到独立，沙阿博士制定了一个全面的职业发展计划，充分利用费城儿童医院和宾夕法尼亚大学丰富的研究，教育和协作环境。Shah博士的职业规划包括在转化研究，遗传学，免疫学和生物信息学的教学课程，重点培训必要的实验室技能和临床研究，科学丰富和专业发展方面通过参加校内和校外，研讨会和会议，并明确界定独立的步骤。她得到了儿科和内分泌科的全力支持，以确保有足够的时间和资源进行培训。此外，她将通过她的导师，Muredach Reilly博士和Karen Teff博士，以及在她提出的研究的关键领域精心挑选的专家咨询委员会获得指导。为了完成她提出的研究，Shah博士将有机会获得宾夕法尼亚大学/CHOP CTSA赞助的临床和转化研究中心（CTRC）的资源，Reilly博士的实验室，核心设施和统计专业知识。通过完成本申请中概述的研究和教育计划，Shah博士将获得必要的经验，以建立一个独立的转化研究计划，探索成人-儿科范围内的肥胖机制。鼠长链ω-3多不饱和脂肪酸（LC n-3 PUFA）在临床上用于治疗血脂异常，并在临床前细胞和啮齿动物模型中证明可减少脂肪细胞和单核细胞炎症，并防止高脂喂养期间脂肪白细胞浸润;然而，缺乏体内人体数据。脂肪炎症和巨噬细胞浸润的关键驱动因素是脂肪趋化因子信号传导，包括CCL 2-CCR 2和CX 3CL 1-CX 3CR 1途径。LC n-3 PUFA在脂肪中的抗炎作用是否是通过这些趋化因子系统介导的，这是由喂食n-3 PUFA的小鼠中CCL 2水平降低所提示的，但尚未具体探索。LC n-3 PUFA在脂肪中发挥抗炎作用的机制也未明确定义。最近，已经鉴定了LC n-3 PUFA受体GPR 120，其在小鼠和细胞模型中似乎调节核因子κ β（NF）信号传导的n-3 PUFA抑制以减少细胞和脂肪组织中的炎症。在人类中，GPR 120功能的丧失会增加肥胖的风险。总之，这些发现提示了LC n-3 PUFA减轻肥胖相关脂肪炎症的潜在机制.我们假设LC n-3 PUFA通过GPR 120抑制NF- B信号传导调节脂肪细胞-巨噬细胞趋化因子信号传导（CCL 2-CCR 2和CX 3CL 1-CX 3CR 1）以减少全身和脂肪炎症。本申请涉及以下假设：1）体外对原代人脂肪细胞的LC n-3 PUFA处理通过GPR 120抑制趋化因子诱导的脂肪细胞炎症和脂肪细胞-单核细胞相互作用2）体外对人单核细胞和巨噬细胞的LC n-3 PUFA处理修饰CX 3CL 1-CX 3CR 1和CCL 2-CCR 2以减少这些白细胞的炎症状态和在GPR 120-CCR 120中与脂肪细胞的所得相互作用。依赖性方式和3）与安慰剂相比，用n-3 PUFA补充剂Lovaza治疗肥胖人类受试者调节CX 3CL 1-CX 3CR 1和CCL 2-CCR 2趋化因子系统以减少炎性白细胞募集到人类脂肪中。在原代人细胞中的体外研究将评估单个n-3 PUFA和GPR 120激动剂对基线和炎症刺激的趋化因子信号传导、CX 3CL 1-CX 3CR 1和CCL 2-CCR 2诱导的单核细胞-脂肪细胞粘附和迁移以及NF活化的影响。GPR 120和NFkB的敲低将用于确定抗炎作用是否由这些途径调节。对于临床试验，肥胖、非糖尿病受试者将接受Lovaza或安慰剂治疗8周;计划在基线和8周时进行访视，以评估胰岛素敏感性、脂肪和血液趋化因子水平、NF信号传导因子以及脂肪和循环中表达CX 3CR 1和CCR 2的巨噬细胞和单核细胞（通过流式细胞术进行定量）。

DESCRIPTION (provided by applicant): Obesity and its resultant complications represent a major public health; further understanding of the underlying pathophysiology will aid the development of preventive and therapeutic modalities. As a pediatric endocrinologist, the P.I. Dr Shah has a committed interest in studying the early mechanistic processes involved in obesity-related metabolic dysregulation, with the hope of exploring novel preventive and treatment options. Her ultimate career goal is to become an independent translational researcher in the field of endocrinology, with a focus in obesity and adipose tissue biology across the pediatric and adult age spectrum. Through her current work with her mentor and training in the Masters in Translational Research program, she has developed essential skills and experience in clinical and translational research. This will allow her to perform the work proposed in this application, which integrates a human clinical trial with studies in human tissues and cells to explore novel pathways in obesity-related adipose inflammation. To acquire further expertise and knowledge as she transitions to independence, Dr Shah has devised a comprehensive career development plan that takes full advantage of the rich research, educational, and collaborative environment at the Children's Hospital of Philadelphia and the University of Pennsylvania. Dr Shah's career plan includes strong mentorship in translational research, didactic coursework in genetics, immunology, and bioinformatics, focused training in necessary laboratory skills and aspects of clinical research, scientific enrichment and professional development through attendance of intra- and extra-mural, seminars and conferences, and clearly delineated steps to independence. She has the full support of the Department of Pediatrics and Division of Endocrinology to ensure adequate time and resources for this training. Additionally, she will receive guidance through her mentors, Dr Muredach Reilly and Dr Karen Teff, and a carefully selected advisory committee of experts in critical areas of her proposed research. In order to complete her proposed studies, Dr Shah will have access to resources in the Penn/CHOP CTSA-sponsored Clinical and Translational Research Center (CTRC), Dr Reilly's laboratory, core facilities, and statistical expertise. Through the completion of the research and educational plans outlined in this application, Dr Shah will gain the experience necessary to establish an independent translational research program exploring mechanisms in obesity across the adult-pediatric spectrum. Murine long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) are used clinically to treat dyslipidemia and are demonstrated in pre-clinical cell and rodent models to decrease adipocyte and monocyte inflammation and prevent adipose leukocyte infiltration during high-fat feeding; in vivo human data, however, is lacking. A key driver of adipose inflammation and macrophage infiltration is adipochemokine signaling, including the CCL2-CCR2 and CX3CL1-CX3CR1 pathways. Whether LC n-3 PUFA anti-inflammatory actions in adipose are mediated via these chemokine systems is suggested by decreased CCL2 levels in mice fed n-3 PUFA but has not specifically been explored, The mechanisms by which LC n-3 PUFA exerts anti-inflammatory effects in adipose are also not clearly defined. Recently a LC n-3 PUFA receptor, GPR120, has been identified that in mouse and cell models appears to modulate n-3 PUFA inhibition of nuclear factor kappa beta (NF signaling to decrease inflammation in cells and adipose tissue. In humans, loss of function of GPR120 confers increased risk of obesity. Together these findings suggest a potential mechanism by which LC n-3 PUFA attenuates obesity- related adipose inflammation. We hypothesize that LC n-3 PUFA modulate adipocyte-macrophage chemokine signaling (CCL2-CCR2 and CX3CL1-CX3CR1) to decrease systemic and adipose inflammation via GPR120 inhibition of NF- B signaling. This application addresses the following hypotheses: 1) LC n-3 PUFA treatment of primary human adipocytes in vitro suppresses chemokine-induced adipocyte inflammation and adipocyte-monocyte interactions via GPR120 2) LC n-3 PUFA treatment of human monocytes and macrophages in vitro modifies CX3CL1-CX3CR1 and CCL2-CCR2 to reduce the inflammatory state of these leukocytes and resultant interactions with adipocytes in a GPR120-dependent manner and 3) treatment of obese human subjects with the n-3 PUFA supplement Lovaza, compared to placebo, regulates the CX3CL1-CX3CR1 and CCL2-CCR2 chemokine systems to reduce inflammatory leukocyte recruitment into human adipose. In vitro studies in primary human cells will assess effects of individual n-3 PUFAs and GPR120 agonists on baseline and inflammatory stimulated chemokine signaling, CX3CL1-CX3CR1 and CCL2-CCR2 induced monocyte-adipocyte adhesion and migration, and.NF activation. Knockdown of GPR120 and NFkB will be used to determine whether anti- inflammatory effects are modulated by these pathways. For the clinical trial, obese, non-diabetic subjects will be treated for 8 weeks with Lovaza or placebo; visits are planned at baseline and 8 weeks for assessment of insulin sensitivity, adipose and blood chemokine levels, NF signaling factors, and adipose and circulating CX3CR1- and CCR2-expressing macrophage and monocyte quantification via flow cytometry.