Omega-3 PUFA suppress adipochemokines in human obesity

描述(由申请人提供)：肥胖及其引起的并发症是一个重大的公共卫生问题；进一步了解其潜在的病理生理学将有助于制定预防和治疗方法。作为一名儿科内分泌学家，P.I.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一名独立翻译研究员，专注于儿童和成人年龄段的肥胖和脂肪组织生物学。通过她目前与导师的工作和翻译研究硕士项目的培训，她在临床和翻译研究方面发展了基本的技能和经验。这将使她能够执行这项申请中提出的工作，该申请将人类临床试验与人类组织和细胞研究相结合，以探索肥胖相关脂肪炎症的新途径。为了在过渡到独立的过程中获得更多的专业知识，沙阿博士设计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、教育和合作环境。沙阿博士的职业计划包括在翻译研究方面提供强有力的指导，在遗传学、免疫学和生物信息学方面进行教学课程，在必要的实验室技能和临床研究方面进行重点培训，通过参加校内外研讨会和会议来丰富科学知识和专业发展，以及明确描述独立的步骤。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这次培训。此外，她将通过她的导师Muredach Reilly博士和Karen Tef博士，以及在她提议的研究的关键领域精心挑选的专家咨询委员会获得指导。为了完成她提议的研究，Shah博士将获得宾夕法尼亚大学/CHOP CTSA赞助的临床和翻译研究中心(CTRC)、Reilly博士的实验室、核心设施和统计专业知识的资源。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换研究计划，探索成人-儿科肥胖的机制。小鼠长链omega-3多不饱和脂肪酸(Lcn-3 PUFA)被临床用于治疗血脂异常，并在临床前细胞和啮齿动物模型中证明在高脂喂养期间可以减少脂肪细胞和单核细胞炎症，防止脂肪白细胞渗透；然而，体内人类数据缺乏。脂肪细胞炎症和巨噬细胞浸润的一个关键驱动因素是脂肪趋化因子信号，包括CCL2-CCR2和CX3CL1-CX3CR1通路。Lcn-3PUFA是否通过这些趋化因子系统介导脂肪中的抗炎作用可能是通过降低摄入n-3PUFA的小鼠的CCL2水平来实现的，但尚未有专门的研究，Lcn-3PUFA发挥脂肪抗炎作用的机制也不清楚。最近，LCn-3PUFA受体GPR120被发现，在小鼠和细胞模型中，似乎调节n-3PUFA抑制核因子kappaβ(NF)信号，从而减轻细胞和脂肪组织的炎症。在人类中，GPR120功能的丧失会增加肥胖的风险。综上所述，这些发现提示了LC n-3多不饱和脂肪酸减轻肥胖相关脂肪炎症的潜在机制。我们推测，LCn-3多不饱和脂肪酸通过调节脂肪细胞-巨噬细胞趋化因子信号(CCL2-CCR2和CX3CL1-CX3CR1)，通过GPR120抑制NF-B信号来减轻全身和脂肪炎症。本应用解决了下列假设：1)通过GPR120体外处理原代人类脂肪细胞的LC n-3PUFA抑制趋化因子诱导的脂肪细胞炎症和脂肪细胞与单核细胞的相互作用2)体外处理人单核细胞和巨噬细胞的LC n-3PUFA通过GPR120依赖的方式修改CX3CL1-CX3CR1和CCL2-CCR2以降低这些白细胞的炎症状态和由此产生的与脂肪细胞的相互作用；3)与安慰剂相比，服用n-3PUFA补充洛瓦扎的肥胖受试者调节CX3CL1-CX3CR1和CCL2-CCR2趋化因子系统，以减少炎性白细胞募集到人体脂肪中。在原代人体细胞中进行的体外研究将评估单独的n-3PUFAs和GPR120激动剂对基线和炎症刺激的趋化因子信号、CX3CL1-CX3CR1和CCL2-CCR2诱导的单核细胞-脂肪细胞黏附和迁移以及NF激活的影响。GPR120和NFkB基因的敲除将被用来确定抗炎作用是否受到这些途径的调节。在临床试验中，肥胖的非糖尿病患者将接受为期8周的Lovaza或安慰剂治疗；访问计划在基线和8周进行，以评估胰岛素敏感性、脂肪和血液趋化因子水平、核因子信号因子以及脂肪和循环中表达CX3CR1和CCR2的巨噬细胞和单核细胞通过流式细胞仪进行定量。

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.