Omega-3 PUFA suppress adipochemokines in human obesity

描述（由申请人提供）：肥胖及其并发症是一项重大的公共健康问题；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，P.I. Shah博士一直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转化研究员，专注于儿童和成人年龄段的肥胖和脂肪组织生物学。通过目前与导师的合作以及在转化研究硕士项目中的培训，她在临床和转化研究方面积累了必要的技能和经验。这将使她能够完成本申请中提出的工作，将人体临床试验与人体组织和细胞研究结合起来，探索与肥胖相关的脂肪炎症的新途径。为了在她过渡到独立时获得更多的专业知识和知识，Shah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。Shah博士的职业规划包括在转化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的教学课程，在必要的实验室技能和临床研究方面的重点培训，通过参加校内和校外研讨会和会议来丰富科学和专业发展，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，她将从她的导师Muredach Reilly博士和Karen Teff博士，以及在她拟议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资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项目，探索成人-儿童肥胖症的机制。小鼠长链omega-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被证明可以减少脂肪细胞和单核细胞炎症，防止高脂肪喂养过程中脂肪白细胞的浸润；然而，缺乏体内的人体数据。脂肪趋化因子信号通路是脂肪炎症和巨噬细胞浸润的关键驱动因素，包括CCL2-CCR2和CX3CL1-CX3CR1通路。lcn -3 PUFA在脂肪中的抗炎作用是否通过这些趋化因子系统介导，可以通过喂食n-3 PUFA的小鼠CCL2水平降低来推测，但尚未具体探讨，lcn -3 PUFA在脂肪中发挥抗炎作用的机制也未明确定义。最近，一种LC n-3 PUFA受体GPR120在小鼠和细胞模型中被发现，它似乎可以调节n-3 PUFA对核因子κ β (NF)信号的抑制，从而减少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。总之，这些发现提示了lcn -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信号传导来减少全身和脂肪炎症。该应用程序解决了以下假设：1) LC n-3 PUFA体外治疗原代人脂肪细胞通过GPR120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)LC n-3 PUFA体外治疗人单核细胞和巨噬细胞修饰CX3CL1-CX3CR1和CCL2-CCR2，以GPR120依赖的方式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，使用n-3 PUFA补充剂Lovaza治疗肥胖受试者。调节CX3CL1-CX3CR1和CCL2-CCR2趋化因子系统，以减少炎性白细胞向人类脂肪的募集。在原代人细胞中的体外研究将评估单个n-3 PUFAs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-脂肪细胞粘附和迁移的影响。NF激活。GPR120和NFkB的下调将用于确定抗炎作用是否受这些途径的调节。在临床试验中，肥胖、非糖尿病受试者将使用Lovaza或安慰剂治疗8周；计划在基线和8周进行随访，评估胰岛素敏感性、脂肪和血液趋化因子水平、NF信号因子、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术定量。

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.