The role of circulating Slit2 in adipose thermogenesis and diabetes

循环 Slit2 在脂肪产热和糖尿病中的作用

• 批准号: 9349495
• 负责人: Katrin Jennifer Svensson
• 金额: $ 9万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-08 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9349495
• 关键词: Address; Adenoviruses; Adipocytes; Adipose tissue; Adrenergic Receptor; Adrenergic beta-Agonists; Adverse effects; Agonist; Alpha Cell; Animals; Award; Binding; Binding Proteins; Biochemical; Biochemistry; Biological; Blood; Body Weight Changes; Brain; Brown Fat; C-terminal; Cell Line; Cell Surface Receptors; Cell surface; Cells; Chronic; Chronically Ill; Clinical; Collection; Communication; Communities; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Diabetes Mellitus; Diet; Disease; Dose; Energy Metabolism; Engineering; Evaluation; Faculty; Fatty acid glycerol esters; Future; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; Gene Expression; Genetic; Genetic Transcription; Glucose tolerance test; Goals; Health Care Costs; Heart Diseases; Heart Rate; High Fat Diet; Homologous Gene; Hormones; Human; Institution; Insulin Resistance; Knock-out; Knockout Mice; Knowledge; Laboratories; Laboratory Research; Leadership; Leptin; Ligands; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mentors; Metabolic; Metabolic Diseases; Metabolism; Methods; Molecular; Monitor; Mus; Nervous system structure; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Occupations; Organ; Oxygen Consumption; Pathway interactions; Pattern; Peripheral; Phase; Physiological; Physiology; Positioning Attribute; Postdoctoral Fellow; Preparation; Process; Protein Fragment; Proteins; Proteomics; Receptor Signaling; Recombinant Proteins; Recombinants; Regulation; Research; Research Institute; Research Personnel; Research Proposals; Resources; Rodent; Role; Scheme; Signal Pathway; Signal Transduction; Small Interfering RNA; Stroke; Subcutaneous Injections; Supervision; Techniques; Testing; Therapeutic; Thermogenesis; Tissues; Training; United States National Institutes of Health; Weight Gain; Weights and Measures; Work; Writing; adiponectin; autocrine; base; beta-adrenergic receptor; blood glucose regulation; career; career development; citrate carrier; clinical translation; diabetes mellitus therapy; diabetic; effective therapy; experimental study; feeding; genomic data; glucose tolerance; improved; in vivo; interest; loss of function; medical schools; mouse model; novel; novel therapeutics; obesity treatment; overexpression; pandemic disease; paracrine; professor; programs; protein purification; receptor; skill acquisition; skills; therapeutic development; therapeutic protein; tumor; uptake

ABSTRACT The increase in obesity worldwide has led to rising health care costs and the number of chronically ill people suffering from obesity-related disorders such as type 2 diabetes. The overarching goal of this research proposal is to better understand brown fat activation and to develop new therapeutic manipulations targeting brown fat to treat metabolic disease such as type 2 diabetes. This work will test the hypothesis that the secreted protein Slit2 is involved in adipose thermogenesis and if administration of the protein could be a safe and effective treatment for diabetes. We have previously utilized unbiased quantitative proteomics to identify novel secreted proteins involved in browning. Using this method, I have identified new mechanisms by which thermogenesis can be activated in adipose tissue. One of the candidates from this approach identified Slit2 as a secreted factor from thermogenic adipocytes. Slit2 had previously been described for its function in brain, and were not believed to be circulating in blood. In addition, I have found a C-terminal cleavage fragment of Slit2, Slit2-C, which has no known peripheral function and acts through an unknown cell-surface receptor. My preliminary data demonstrate that increasing Slit2-C circulating levels in obese, insulin resistant mice improves whole body glucose homeostasis and energy expenditure. These mice show increased oxygen consumption in the brown fat tissue as well as activation of a thermogenic transcriptional program. At least in part, Slit2-C acts through activation of the canonical PKA pathway. In the mentored phase of the award, Aim 1 will test the hypothesis that Slit2 is required for regulating adipose tissue thermogenesis in vivo by performing metabolic characterization of the adipocyte-specific knockout of Slit2. In aim 2 of the mentored phase, I will test the hypothesis that administration of Slit2-C recombinant protein to diabetic rodents will improve diet-induced insulin resistance. Aim 3 will be conducted in the independent phase and will use multiple approaches to determine the functional receptor and signaling pathways for Slit2-C with further potential for clinical translation. I will use a combination of animal physiology and genetics, biochemical protein purification, and mass spectrometry techniques to address the questions in the proposal. If successful, I anticipate that the findings in this proposal has the potential to contribute with new treatments for type-2 diabetes. My current and long-term career objectives are to identify pathways involved in adipose tissue metabolism and to develop new protein therapeutics that regulates glucose homeostasis and has the possibility to improve diabetes. I have a longstanding interest in studying ligand-receptor interactions, macromolecular uptake and mechanisms of intracellular signaling in tumor development. The findings in this proposal are directly building upon my discovery in my postdoctoral work and would be completed in the defined award period. Therefore, study of secreted factor Slit2-C as a new protein therapy for diabetes is a logical extension of my research. My career trajectory after the mentored phase of the K99 is to become an assistant professor at a leading academic research institute. Dr. Bruce Spiegelman, a well-recognized leader in the field of brown fat and diabetes will mentor my scientific and career development. Dr. Spiegelman has successfully trained numerous postdoctoral fellows now holding faculty positions in academic institutions. The Spiegelman laboratory and Harvard Medical School research community provide an ideal setting for training future independent investigators. My plan for career activities includes a continous progress evaluation with my mentor, training in biochemistry and protein therapies, supervision and leadership training, educational coursework, and mentored job search. These career activities will be imperative in the preparation to establish my own research laboratory. In the independent phase, I plan to leverage my biochemical skills and development of new protein biologics and signaling pathways to scientifically separate myself from my mentor. My long-term scientific goal is to establish myself as an independent researcher to drive the field of metabolism and diabetes forward. My focus will be to study biological pathways of circulating hormones and to further study adrenergic receptor-independent pathways of thermogenesis activation in the context of obesity and diabetes. Together with the outstanding resources at Harvard Medical School, this will maximize my potential to successfully transition to independence. The NIH Pathway to Independence Award will be essential for my transition because it will enable me to gain additional training in mouse models of obesity and protein therapies as well as improving my writing and communication skills required for a successful transition.

抽象的 全球肥胖人数的增加导致医疗保健费用和慢性病患者数量上升 患有与肥胖相关的疾病，例如 2 型糖尿病。本研究的总体目标 建议是为了更好地了解棕色脂肪的激活并开发新的治疗方法 棕色脂肪可治疗代谢性疾病，例如 2 型糖尿病。这项工作将检验以下假设： 分泌蛋白 Slit2 参与脂肪生热作用，如果施用该蛋白可能是安全的 以及糖尿病的有效治疗。我们之前利用无偏定量蛋白质组学来识别 参与褐变的新型分泌蛋白。使用这种方法，我已经确定了新的机制 脂肪组织中的产热作用可以被激活。该方法的一位候选者将 Slit2 确定为 产热脂肪细胞分泌的因子。 Slit2 之前曾被描述过其在大脑中的功能， 并且据信不会在血液中循环。另外，我还发现了一个C端切割片段 Slit2，Slit2-C，没有已知的外周功能，通过未知的细胞表面受体发挥作用。我的 初步数据表明，增加肥胖、胰岛素抵抗小鼠的 Slit2-C 循环水平可改善 全身葡萄糖稳态和能量消耗。这些小鼠的耗氧量增加 棕色脂肪组织以及生热转录程序的激活。 Slit2-C 至少在一定程度上起作用 通过激活经典 PKA 途径。在该奖项的指导阶段，Aim 1 将测试 假设 Slit2 是通过执行代谢来调节体内脂肪组织产热所必需的 Slit2 脂肪细胞特异性敲除的表征。在指导阶段的目标 2 中，我将测试 假设对糖尿病啮齿动物施用 Slit2-C 重组蛋白将改善饮食引起的糖尿病 胰岛素抵抗。目标 3 将在独立阶段进行，并将使用多种方法来实现 确定 Slit2-C 的功能受体和信号通路，具有进一步的临床潜力 翻译。我将结合动物生理学和遗传学、生化蛋白质纯化以及 质谱技术来解决提案中的问题。如果成功的话，我预计 该提案中的发现有可能为 2 型糖尿病的新疗法做出贡献。 我当前和长期的职业目标是确定脂肪组织中涉及的途径 代谢并开发新的蛋白质疗法来调节葡萄糖稳态并具有 改善糖尿病的可能性。我长期以来对研究配体-受体相互作用很感兴趣， 肿瘤发展中的大分子摄取和细胞内信号传导机制。本次调查结果 该提案直接建立在我博士后工作中的发现之上，并将于 明确的奖励期限。因此，分泌因子Slit2-C作为糖尿病新蛋白疗法的研究是一个新的课题。 我的研究的逻辑延伸。在 K99 的指导阶段之后，我的职业轨迹是成为一名 领先学术研究机构的助理教授。布鲁斯·斯皮格曼 (Bruce Spiegelman) 博士是公认的医学领域的领导者 棕色脂肪和糖尿病领域将指导我的科学和职业发展。斯皮格曼博士有 成功培养了众多博士后研究员，目前在学术机构担任教职。这 斯皮格曼实验室和哈佛医学院研究社区为培训提供了理想的环境 未来的独立调查员。我的职业活动计划包括与我的同事进行持续的进展评估 导师、生物化学和蛋白质疗法培训、监督和领导力培训、教育 课程作业和指导求职。这些职业活动对于准备建立 我自己的研究实验室。在独立阶段，我计划利用我的生化技能和 开发新的蛋白质生物制剂和信号通路，以科学地将自己与导师分开。 我的长期科学目标是让自己成为一名独立研究人员，推动新陈代谢领域的发展 和糖尿病向前发展。我的重点是研究循环激素的生物途径并进一步 研究肥胖背景下肾上腺素能受体独立的产热激活途径 糖尿病。再加上哈佛医学院的优秀资源，这将最大限度地发挥我的潜力 成功过渡到独立。 NIH 独立之路奖对于我来说至关重要 过渡，因为它将使我能够获得肥胖小鼠模型和蛋白质疗法的额外培训 以及提高成功过渡所需的写作和沟通技巧。

项目成果

Discovery of Hydrolysis-Resistant Isoindoline N-Acyl Amino Acid Analogues that Stimulate Mitochondrial Respiration.

发现刺激线粒体呼吸的抗水解异吲哚啉 N-酰基氨基酸类似物。

• DOI: 10.1021/acs.jmedchem.8b00029
• 发表时间: 2018-04-12
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Lin H;Long JZ;Roche AM;Svensson KJ;Dou FY;Chang MR;Strutzenberg T;Ruiz C;Cameron MD;Novick SJ;Berdan CA;Louie SM;Nomura DK;Spiegelman BM;Griffin PR;Kamenecka TM
• 通讯作者: Kamenecka TM