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测定方法，以鉴定促进人类细胞中过氧化物酶体生物发生的化合物。我们鉴定的化合物可以成为有价值的化学探针，用于模拟过氧化物酶体的生物发生障碍，并评估过氧化物酶体对动物模型中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）的治疗潜力。最初的分析开发将以高含量/基于图像的筛选格式进行，目的是将分析转换为分子文库生产中心网络（MLPCN）。过氧化物酶体是脂肪酸氧化的主要位点，也是细胞中长链脂肪酸（VLCFA）氧化的唯一位点。增加人类细胞过氧化物酶体生物发生的化合物可能为ii型糖尿病和代谢综合征提供新的治疗途径。血浆脂肪酸增加有助于胰岛素抵抗和高血糖，这是代谢综合征和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，特别是肝脏（“脂肪肝”）的异位脂质积累，从而损害其功能。因此，通过增加脂肪酸氧化来防止非脂肪组织的脂质积累和减少血浆中的脂肪酸可能是代谢紊乱的治疗方法。结构上不相关的化合物增加过氧化物酶体的生物发生，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。我们将使用一种高含量的筛选试验，以确定使用独立于直接PPAR激活的机制促进人类细胞中过氧化物酶体生物发生的化合物（非经典过氧化物酶体生物发生）。我们鉴定的化合物将通过多次正交二次试验进行反筛选，以确认细胞培养中过氧化物酶体功能的增加。基因表达谱将与刺激过氧化物酶体生物发生的化合物进行，以确定不同的机制。本研究确定的化学探针可用于模拟过氧化物酶体生物发生障碍，并在动物模型中评估过氧化物酶体对正常代谢和生理的贡献。分析开发将以高含量/基于图像的筛选格式进行，目的是将分析转移到分子库生产中心网络（MLPCN）高含量专业筛选中心（Burnham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).