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

描述（由申请人提供）：我们将优化高含量测定，以鉴定促进人类细胞中过氧化物酶体生物合成的化合物。我们确定的化合物可以是有价值的化学探针，用于模拟过氧化物酶体生物合成障碍和评估过氧化物酶体对动物模型中正常代谢和生理学的贡献。此外，化合物可具有治疗代谢综合征（MetS）和2型糖尿病（T2 D）的治疗潜力。将以基于高内容/图像的筛选形式进行初始检测开发，目的是将检测转化为分子库生产中心网络（MLPCN）。过氧化物酶体代表脂肪酸氧化的主要位点，并且是细胞中极长链脂肪酸（VLCFA）氧化的唯一位点。增加人类细胞中过氧化物酶体生物合成的化合物可能为II型糖尿病和代谢综合征提供新的治疗途径。增加的血浆脂肪酸有助于胰岛素抵抗和高血糖症，这是代谢综合征和T2 D的风险因素。脂毒性假说表明，血浆脂肪酸升高可能导致非脂肪组织，特别是肝脏（“脂肪肝”）中的异位脂质积聚，这将损害其功能。因此，通过增加脂肪酸氧化来防止非脂肪组织中的脂质积聚和减少血浆中的脂肪酸可以提供代谢紊乱的治疗。增加过氧化物酶体生物合成的结构无关化合物确实可以改善啮齿动物的代谢综合征和糖尿病症状，从而验证了这一表型目标。我们将使用高含量筛选测定来鉴定使用独立于直接PPAR活化的机制（非经典过氧化物酶体生物发生）促进人细胞中过氧化物酶体生物发生的化合物。我们鉴定的化合物将用多个正交二级测定进行反筛选，以确认细胞培养物中过氧化物酶体功能的增加。将使用刺激过氧化物酶体生物发生的化合物进行基因表达谱分析，以确定不同的机制。在这项研究中确定的化学探针可用于建模过氧化物酶体生物合成障碍和评估的贡献过氧化物酶体的正常代谢和生理的动物模型。将以高含量/基于图像的筛选形式进行检测开发，目的是将检测转移至分子库生产中心网络（MLPCN）高含量专业筛选中心（Burnham Institute）。

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).