Targeting a Novel Pocket on ITGAV

项目摘要。美国癌症死亡人数保持在历史最高水平，临床需求迫切 开发新的治疗策略来帮助患者。缺乏有效的治疗方法的部分原因是 目前的科学方法才刚刚开始揭示癌症的潜在复杂性。工艺性 进步正在迅速改变科学发现的格局；例如， 数学建模与基于实验室的验证相结合，可实现更好的组合疗法 治疗癌症。为此，我建议对F99/K00博士后和F99/K00博士后进行培训 过渡奖。对于F99阶段的论文研究，我将重点放在基于实验室的研究技巧上 确定并提出一种治疗癌症的新疗法。在以约580为目标的高级CRISPR屏幕中 我们发现整合素αV(ITGAV)对实体瘤的生存是必不可少的 (结肠癌、胰腺癌和乳腺癌)。为了验证ITGAV是最重要的整合素，我们设计了第二个整合素 对所有26个整合素进行分层筛选，发现ITGAV和整合素Beta 5(ITGB5)是唯一必需的 实体肿瘤中的整合素。有趣的是，整合素必须在α和β之间形成专有的异二聚体。 其中ITGAV和ITGB5是已知的24个亚基之一。作为更基本的配对，ITGAV被探索 高密度CRISPR平铺扫描，我们发现一个小口袋对于ITGAV功能是必不可少的，它是 可修改为小分子结合。基于结构的分析发现了ITGAV的β对的环状结构 与发现的口袋相互作用，导致我们假设口袋对杂二聚体是必不可少的 ITGAV和它的Beta对之间的稳定性。事实上，从500个小分子的高通量筛选中，我们 发现一种化合物似乎结合在我们的口袋里，破坏了ITGAV和ITGB5之间的异源二聚体。 对这一潜力的进一步验证将是论文研究和 完成，将填补未满足的临床需求，因为没有FDA批准的针对整合素的药物获得批准 用于癌症的适应症。为了进一步提高治疗癌症的潜力，我计划使用数学建模 通过了解癌症信号的复杂性来确定新的治疗策略的方法 K00阶段，拟开展博士后工作。研究复杂的癌症信号，与Dr。 Pirrotte，我们在ITGAV被敲除的细胞中产生了激酶活性分数。有了这些数据，我们就可以建模 信号的影响，因为它与癌细胞中可测量的变化有关。具体地说，我们将研究细胞 周期控制，这与ITGAV损耗一起被抑制。此外，我们可以将已知的抑制剂模拟为常见的 信号级联作为新的组合治疗策略。为了确认我们的模型，我将使用实验室 基于在F99阶段发展的技能。总的来说，通过F99/K00奖的培训，我将获得以下技能 能够建立研究癌症的数学模型，并用基于实验室的技能验证这些模型。 这将使我成为独立研究和翻译研究的领先科学家.

Project Summary. Cancer deaths remain at an all-time high in the United States leaving an urgent clinical need to develop novel therapeutic strategies to help patients. The lack of effective treatments is in part due to underlying complexities in cancer that current scientific approaches are just beginning to uncover. Technological advances are rapidly changing the landscape of scientific discovery; for example, the combination of mathematical modeling in tandem with laboratory based validation leading to better combinational therapies to treat cancer. For this reason, I propose training in both with the F99/K00 Predoctoral to Postdoctoral Fellow Transition Award. For the F99 phase, the dissertation research, I will focus on laboratory based research skills to identify and propose a novel therapeutic to treat cancers. In a high level CRISPR screen targeting about 580 genes on the cell surface we found that Integrin Alpha V (ITGAV) is essential for the survival of solid tumors (colon, pancreatic, and breast cancer). To validate ITGAV as the most essential integrin we designed a second layer screen targeting all 26 integrins and found that ITGAV and Integrin Beta 5 (ITGB5) are the only essential integrins in solid tumors. Interestingly, integrins must for an obligate heterodimer between an alpha and a beta subunit of which ITGAV and ITGB5 are one of the known 24. As the more essential pair, ITGAV was probed with a high-density CRISPR tiling scan and we found a small pocket to be essential for ITGAV function and it was amendable to small molecule binding. A structure based analysis found a loop structure of the beta pair of ITGAV interacts with the discovered pocket, leading to our hypothesis that the pocket is essential for the heterodimer stability between ITGAV and its beta pair. Indeed, from a high-throughput screen of 500 small molecules we found one compound that appears to bind in our pocket and disrupt the heterodimer between ITGAV and ITGB5. Further validation of this potential will be the remaining work to be done for the dissertation research and upon completion, will fill an unmet clinical need since no there no FDA approved drugs targeting integrins approved for cancer indications. To further advance the potential to treat cancer I plan to use mathematical modeling approaches to identify novel therapeutic strategies by understanding the complexities of cancer signaling during the K00 phase, the proposed postdoctoral work. To study complex cancer signaling, in collaboration with Dr. Pirrotte, we generated kinase activity scores in cells where ITGAV was knocked out. With this data we can model the effects of signaling as it relates to measurable changes in the cancer cells. Specifically, we will study cell cycle control, which is inhibited with ITGAV loss. Additionally, we can model known inhibitors to common signaling cascades as novel combinational therapeutic strategies. To confirm our model, I will use laboratory based skill developed during the F99 phase. Overall with the training with the F99/K00 award I will gain skills to be able to build mathematical models to study cancer and validate those models with laboratory based skills. This will allow me to become and independent research and leading scientist in translational research.