Developing the first TPI Df therapeutics

摘要/项目摘要： TPI DF是一种毁灭性的、无法治疗的儿童代谢性疾病，会导致贫血、瘫痪、不可逆转 脑损伤和过早死亡。磷酸丙糖中的多种单一氨基酸取代 异构酶(TPI)是一种致病物质，可导致快速发展的多系统疾病。重要的是，所有 已知的致病TPI DF突变导致一种保留功能的蛋白质，其发病机制已知 这是由于蛋白质质量控制途径(PQC)增加了功能蛋白质的周转。我们 我开发了一种基于E104D突变的细胞模型的人类细胞TPI DF检测方法 并将其应用于高内容、高通量成像。我们已经在试点屏幕中使用了该模型 并验证了其在识别调节人类细胞中突变TPI蛋白水平的新化合物方面的有效性。 我们建议将化验方法发展为完全的HTS标准，对几种相关化合物进行筛选 图书馆，并确定一流的TPI DF小分子疗法。我们将验证辅助数据库中的命中率 TPI DF患者细胞中TPI稳定性和活性的测定，在体外毒理学小组中优先考虑 和代谢稳定性分析，检查先导化合物的结构活性关系(SARS) 使用最近开发的小鼠TPI DF模型在体内实质上验证了它们。总体而言，该项目将 发现并验证有史以来第一种治疗TPI DF的方法，这将为临床试验提供基础。

Abstract/Project summary: TPI Df is a devastating untreatable childhood metabolic disease resulting in anemia, paralysis, irreversible brain damage and premature death. Numerous single amino acid substitutions in Triosephosphate Isomerase (TPI) are pathogenic and result in rapidly progressing multisystem disease. Importantly, all known pathogenic TPI Df mutations result in a protein that retains function and pathogenesis is known to result from increased turnover of the functioning protein by Protein Quality Control pathways (PQC). We have developed a human cellular TPI Df assay based on a cellular model of the “common” E104D mutation and implemented it for high-content, high-throughput imaging. We have used this model in a pilot screen and validated its utility to identify novel compounds that modulate mutant TPI protein levels in human cells. We propose to develop the assay to full HTS standards, conduct a screen of several relevant compound libraries, and identify first-in-class TPI Df small molecule therapies. We will validate hits in secondary assays for TPI stability and activity in TPI Df patient cells, prioritize them in a panel of in vitro toxicology and metabolic stability assays, examine structure activity relationships (SARs) of the lead compounds and substantially validate them in vivo using a recently developed mouse TPI Df model. Overall this project will discover and validate the first ever treatments for TPI Df that will provide the basis for clinical trials.