HTS for Modulators of MitoNEET Proteins' Function

描述（由申请人提供）：全球肥胖症的增加与肥胖相关疾病的急剧增加相平行，最明显的是II型糖尿病。据预测，这种疾病在今后几十年内将达到流行病的程度。因此，了解II型糖尿病的生化过程并确定治疗干预的新目标对国家和世界健康至关重要。治疗II型糖尿病的一些最广泛处方的胰岛素增敏药物属于噻唑烷二酮（TZD）类分子。发现这些药物还减少许多与代谢综合征相关的病理，包括高血压、腹部肥胖、冠状动脉炎症、多发性硬化、阿尔茨海默病和肌萎缩侧索硬化。虽然TZDs最初被认为仅通过激活核转录因子PPAR发挥其作用，但目前已知许多有益作用是以不依赖PPAR的方式介导的。TZDs最近被证明与一种名为mitoNEET的新型线粒体蛋白质靶点相互作用。我们报道了蛋白mitoNEET是一个氧化还原活性，pH不稳定的2Fe-2S簇包含在线粒体外膜蛋白。这是线粒体外膜中唯一已知的Fe-S蛋白。Miner 1是mitoNEET的内质网同源物，在维持健康和长寿方面很重要，并与癌症和神经退行性疾病相关的蛋白质相互作用。这些蛋白质已经成为从糖尿病到阿尔茨海默氏症等疾病的重要新治疗靶点。该提案的重点是实施高通量筛选方法，以首先鉴定调节该新型蛋白质家族活性的小分子，其次验证二级筛选方法中鉴定的靶标，使用从这些研究中获得的知识开发潜在的治疗剂并测试鉴定的分子在细胞测定中的作用。

DESCRIPTION (provided by applicant): The rise in obesity worldwide parallels a dramatic increase in obesity-associated diseases, most notably type-II diabetes. This disease is predicted to reach epidemic proportions in the next several decades. Thus, understanding the biochemical processes underlying type-II diabetes and identifying new targets for therapeutic intervention are critical for national and world health. Some of the most widely prescribed insulin-sensitizing drugs to treat type-II diabetes belong to the thiazolidinedione (TZD) class of molecules. These drugs were found to also reduce many of the pathologies related to metabolic syndrome including hypertension, abdominal obesity, coronary artery inflammation, multiple sclerosis, Alzheimer's disease and Amyotrophic lateral sclerosis. While the TZDs were originally thought to exert their effects solely through activation of the nuclear transcription factor PPAR¿, it is nw known that many of the beneficial effects are mediated in a PPAR¿-independent manner. The TZDs were recently shown to interact with a novel mitochondrial protein target called mitoNEET. We reported that the protein mitoNEET is a redox-active, pH-labile 2Fe-2S cluster containing protein in the outer mitochondrial membrane. This is the only known Fe-S protein in the outer- mitochondrial membrane. Miner1, an endoplasmic reticulum homolog of mitoNEET, is important in maintaining health and longevity and interacts with proteins associated with cancer as well as neurodegenerative diseases. These proteins have emerged as important new therapeutic targets in diseases ranging from diabetes to Alzheimer's. The focus of this proposal is to implement high through-put screening methods to first identify small molecules that modulate the activity of this novel protein family, second validate the identified targets in secondary screening methods, use the knowledge gained from these studies to develop potential therapeutics and test the effects of identified molecules in cellular assays.