High Throughput Assays to Identify Inhibitors of SUMO-mediated Protein-Protein In

生物医学研究中最大的挑战之一是为新的分子靶点设计配体，以便更好地了解危及生命的疾病的分子机制，并开发创新的，更有效的治疗方法。令人遗憾的是，沿着这条路线的进展沿着缓慢，在食品和药物管理局每年批准的大约50种新药中，只有3 - 5种针对新的分子靶点。SUMO（小泛素样修饰物）是新发现的在治疗癌症和病毒感染中具有重要性的靶点之一，迄今为止尚未开发出针对其的抑制剂。SUMO蛋白家族的蛋白质修饰调节基因转录、细胞周期进程和DNA修复等基本功能。我们的初步数据表明，抑制SUMO介导的蛋白质-蛋白质相互作用可以抑制DNA修复，并使癌细胞对化疗和放疗敏感。该提案的总体目标是开发高通量筛选（HTS）测定，以鉴定SUMO依赖性蛋白质-蛋白质相互作用的抑制剂。同源泛素-蛋白酶体途径的抑制剂在泛素依赖性降解的研究中至关重要，并且作为抗癌治疗剂也非常成功。SUMO依赖性蛋白质-蛋白质相互作用的抑制剂具有类似的前景，并可能导致建立治疗人类疾病的新范式。我们建议使用AlphaScreen技术、荧光共振能量转移和基于细胞的高含量成像方法来开发HTS测定以鉴定此类抑制剂。这些检测试剂盒将由分子库生产中心网络（MLPCN）用于自动化高通量筛选（HTS）。我们还将使用细胞和动物模型进行研究，以确定从HTS中鉴定的先导抑制剂的研究和治疗潜力。 公共卫生相关性：该提案的总体目标是开发高通量筛选测定（HTS），以鉴定SUMO介导的蛋白质-蛋白质相互作用的抑制剂。

DESCRIPTION (provided by applicant): One of the biggest challenges in biomedical research is to design ligands for novel molecular targets so as to better understand the molecular mechanisms of life-threatening diseases and develop innovative, more effective treatments. Regrettably, progress along this line has been slow / among the approximately 50 new drugs approved by the Food and Drug Administration each year, only 3 - 5 address a new molecular target. SUMO (Small Ubiquitin-like Modifier) is one of the newly identified targets with importance in treating cancer and viral infection, for which inhibitors have not been developed to date. Protein modifications by the SUMO family of proteins regulate essential functions such as gene transcription, cell cycle progression and DNA repair. Our preliminary data has shown that inhibition of SUMO-mediated protein-protein interactions can inhibit DNA repair and sensitize cancer cells to chemotherapy and radiation. The overall goal of the proposal is to develop high throughput screening (HTS) assays to identify inhibitors for SUMO-dependent protein-protein interactions. Inhibitors of the homologous ubiquitin-proteosome pathway have been critical in studies of ubiquitin- dependent degradation and have also been extremely successful as anti-cancer therapeutics. Inhibitors of SUMO-dependent protein-protein interactions hold similar promise and could lead to the establishment of a new paradigm in the treatment of human diseases. We propose to develop HTS assays using the AlphaScreen technology, fluorescence resonance energy transfer and cell- based high content imaging approaches to identify such inhibitors. These assays will be used in automated high throughput screening (HTS) by the Molecular Libraries Production Centers Network (MLPCN). We will also perform studies using cellular and animal models to determine the research and therapeutic potentials of the lead inhibitors identified from HTS. PUBLIC HEALTH RELEVANCE: The overall goal of this proposal is to develop high throughput screening assays (HTS) to identify inhibitors for SUMO-mediated protein-protein interactions.