Targeting the RNA Exosome for Cancer Therapeutics

RNA外切体是一种RNA降解机制，通过以下方式在RNA质量控制中发挥关键作用 降解错误或错误拼接的RNA。在许多癌症中，例如胰腺癌，剪接 过程受到损害，导致大量异常转录本的产生。我们假设 癌细胞独一无二地依赖于RNA外切体的活性来去除过量的 因此，RNA外切体可能是一个有影响力的新靶点 胰腺癌的治疗干预。我们已经开发出一套一流的小分子 RNA外切体的抑制剂，并将在这项提议中提炼这些化合物并评估它们的肿瘤- 胰腺癌靶向疗效的三个具体目标。在目标1中，一个领先的优化活动将是 我们首创的一流的RNA外切体抑制剂。将对精制化合物的RNA进行评估 使用体外和基于细胞的RNA外切体活性分析的外切体靶向活性，我们将 识别他们的细胞目标。在目标2中，10种最大的RNA外切体抑制剂的效力将通过以下方式进行评估 通过四甲基偶氮唑盐和克隆存活试验选择最适合体内研究的三种化合物。这前三名 然后，化合物将与电离辐射或吉西他滨结合使用，以评估协同作用。 可诱导的shrna和生长素诱导的蛋白质降解技术将被用来灭活rna。 Exosome和这些系统将与排名前三的抑制剂进行比较，以探索基本机制 RNA外切体在转录和转录后事件中的作用。在目标3中，我们将测试3 胰腺癌PDX原位模型中最有希望单独或联合应用的化合物 吉西他滨和那巴紫杉醇联合治疗方案。这是一个探索一种新癌症的非常创新的提议 治疗目标，并高度关注胰腺癌新的有效治疗方法的开发。 在成功完成这一授权期后，我们打算对最好的 候选化合物。

The RNA exosome is a RNA degradation machinery that plays a critical role in RNA quality control by degrading erroneous or incorrectly spliced RNAs. In many cancers, such as pancreatic cancer, the splicing process is compromised, resulting in the generation of large amounts of aberrant transcripts. We hypothesize that cancer cells are uniquely dependent on RNA exosome activity to remove excess amounts of aberrant transcripts and that the RNA exosome therefore may be an impactful new target for therapeutic intervention for pancreatic cancer. We have developed a set of first-in-class small molecule inhibitors of the RNA exosome and will in this proposal refine these compounds and assess their tumor- targeting efficacy for pancreatic cancer in three specific Aims. In Aim 1 a lead optimization campaign will be initiated of our first-in-class RNA exosome inhibitors. The refined compounds will be evaluated for their RNA exosome-targeting activities using both in vitro and cell-based RNA exosome activity assays and we will identify their cellular targets. In Aim 2, the potencies of the 10 top RNA exosome inhibitors will be assessed by MTT and clonogenic survival assays to select the top three compounds for in vivo studies. These top three compounds will then be used in combination with either ionizing radiation or gemcitabine to assess synergy. Inducible shRNA and auxin-inducible protein degradation technologies will be used to inactivate the RNA exosome and these systems will be compared with the top three inhibitors to explore fundamental mechanisms of action of the RNA exosome in transcriptional and post-transcriptional events. In Aim 3, we will test the 3 most promising compounds in a PDX orthotopic model for pancreatic cancer either alone or in combination regimens with gemcitabine and nab-paclitaxel. This is a very innovative proposal exploring a novel cancer therapeutic target and highly focused on the development of new impactful treatments for pancreatic cancer. Upon the successful completion of this grant period, we intend to perform IND-enabling studies of the best candidate compounds.