High-Content Screening for Peroxisome Biogenesis for Type-II Diabetes.

描述(由申请人提供)：我们将优化一种高含量的分析方法，以确定促进人类细胞中过氧化物酶生物发生的化合物。我们确定的化合物可以作为有价值的化学探针，用于在动物模型中模拟过氧酶体生物发生障碍和评估过氧酶体对正常代谢和生理的贡献。此外，化合物可能具有治疗代谢综合征(METS)和2型糖尿病(T2D)的潜力。最初的化验开发将以高含量/基于图像的筛选格式进行，目的是将化验转换到分子文库生产中心网络(MLPCN)。过氧体是细胞内脂肪酸氧化的主要部位，也是超长链脂肪酸氧化的唯一部位。增加人类细胞中的过氧化物体生物生成的化合物可能为II型糖尿病和代谢综合征提供一种新的治疗途径。血浆脂肪酸增加会导致胰岛素抵抗和高血糖，这些都是代谢综合征和T2D的危险因素。脂毒性假说认为，血浆脂肪酸升高可能导致非脂肪组织异位脂肪堆积，特别是在肝脏(“脂肪肝”)，这将损害其功能。因此，防止非脂肪组织中的脂肪堆积，通过增加脂肪酸氧化来降低血浆中的脂肪酸，可以为代谢紊乱提供治疗。结构上无关的化合物，增加了过氧化物体的生物发生，确实改善了啮齿动物的代谢综合征和糖尿病症状，验证了这一表型靶点。我们将使用高含量的筛选试验，利用独立于PPAR直接激活的机制(非经典的过氧化物体生物发生)来鉴定促进人类细胞中的过氧化物体生物发生的化合物。我们确定的化合物将用多个正交二次分析进行反筛选，以确认细胞培养中增加的过氧化物体功能。基因表达谱将与刺激过氧化物体生物发生的化合物一起进行，以确定不同的机制。本研究中确定的化学探针可用于模拟过氧酶体生物发生障碍，并在动物模型中评估过氧酶体对正常代谢和生理的贡献。检测开发将以高含量/基于图像的筛选格式进行，目的是将检测转移到分子图书馆生产中心网络(MLPCN)高含量专业筛选中心(伯纳姆研究所)。

DESCRIPTION (provided by applicant): We will optimize a high-content assay to identify compounds that promote peroxisome biogenesis in human cells. Compounds we identify can be valuable chemical probes for modeling peroxisome biogenesis disorders and evaluating the contribution of peroxisomes to normal metabolism and physiology in animal models. In addition, compounds may have therapeutic potential for the treatment of metabolic syndrome (MetS) and type 2 diabetes (T2D). The initial assay development will be performed in a high-content/image based screening format, with the intent of translating the assay to the Molecular Libraries Production Centers Network (MLPCN). Peroxisomes represent a major site of fatty acid -oxidation and the only site of very long-chain fatty acid (VLCFA) -oxidation in the cell. Compounds that increase peroxisome biogenesis in human cells might provide a novel therapeutic avenue for type-II diabetes and metabolic syndrome. Increased plasma fatty acids contribute to insulin resistance and hyperglycemia, which are risk factors for both metabolic syndrome and T2D. The lipotoxicity hypothesis suggests that elevated plasma fatty acids may lead to ectopic lipid accumulation in non- adipose tissue, particularly in the liver ("fatty liver"), which would impair its function. Therefore, preventing lipid accumulation in non-adipose tissue and decreasing fatty acids in plasma by increasing fatty acid oxidation could provide treatments for metabolic disorders. Structurally unrelated compounds that increase peroxisome biogenesis do improve metabolic syndrome and diabetic symptoms in rodents, validating this phenotypic target. We will use a high- content screening assay to identify compounds that promote peroxisome biogenesis in human cells using mechanisms independent of direct PPAR activation (non-classical peroxisomal biogenesis). Compounds we identify will be counter-screened with multiple orthogonal secondary assays to confirm increased peroxisomal functionality in cell culture. Gene expression profiling will be performed with compounds stimulating peroxisome biogenesis to identify distinct mechanisms. Chemical probes identified in this study can be used for modeling peroxisome biogenesis disorders and evaluating the contribution of peroxisomes to normal metabolism and physiology in animal models. The assay development will be performed in a high-content/image based screening format, with the intent of transferring the assay to the Molecular Libraries Production Centers Network (MLPCN) high-content specialized screening center (Burnham Institute).