Targeting a Novel Pocket on ITGAV

瞄准 ITGAV 上的新颖口袋

• 批准号: 10529026
• 负责人: NICOLE MATTSON
• 金额: $ 3.23万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-03 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10529026
• 关键词: Acute Lymphocytic Leukemia; Acute Myelocytic Leukemia; Apoptosis; Award; Binding; Biological; Biological Assay; Biology; Breast; CRISPR screen; Cancer Model; Cause of Death; Cell Cycle; Cell Cycle Regulation; Cell Death; Cell surface; Cells; Cessation of life; Cities; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Collaborations; Colon; Colon Carcinoma; Combined Modality Therapy; Complex; Computer Models; Cutaneous T-cell lymphoma; Data; Docking; Drug Targeting; Energy Transfer; Essential Genes; FDA approved; Fellowship; Genes; Goals; Impairment; In Vitro; Integrin alphaV; Integrin beta Chains; Integrins; Knock-in; Knock-out; Laboratories; Lead; Liquid substance; Malignant Neoplasms; Malignant neoplasm of pancreas; Mass Spectrum Analysis; Measurable; Methods; Mitogen-Activated Protein Kinases; Modeling; Monitor; Mutate; Mutation; Oncogenic; Pancreas; Patients; Pharmaceutical Preparations; Phase; Phosphotransferases; Play; Postdoctoral Fellow; Proteomics; Research; Research Project Grants; Role; Scanning; Scientist; Signal Pathway; Signal Transduction; Solid; Solid Neoplasm; Structure; System; Techniques; Therapeutic; Time; Training; Translational Research; United States; Validation; Western Blotting; Work; base; beta catenin; biomarker discovery; cancer cell; cancer survival; cell killing; density; design; effective therapy; high throughput screening; improved; in silico; inhibitor; lead candidate; malignant breast neoplasm; mathematical model; migration; nemo-like kinase; new therapeutic target; novel; novel therapeutic intervention; novel therapeutics; pre-doctoral; predictive modeling; prevent; skills; small molecule; triple-negative invasive breast carcinoma; tumor

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.

项目摘要。美国癌症死亡人数仍处于历史最高水平，临床需求迫切 开发新的治疗策略来帮助患者。缺乏有效治疗的部分原因是 当前的科学方法刚刚开始揭示癌症的潜在复杂性。技术性 进步正在迅速改变科学发现的面貌；例如，组合 数学建模与基于实验室的验证相结合，可带来更好的组合疗法 治疗癌症。因此，我建议对 F99/K00 博士前到博士后进行这两种培训 过渡奖。对于F99阶段的论文研究，我将重点关注基于实验室的研究技能 确定并提出一种治疗癌症的新疗法。在针对约 580 的高水平 CRISPR 筛选中 我们发现细胞表面的基因整合素αV（ITGAV）对于实体瘤的生存至关重要 （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 针对所有 26 个整合素的层筛选，发现 ITGAV 和整合素 Beta 5 (ITGB5) 是唯一必需的 实体瘤中的整合素。有趣的是，整合素必须形成介于 α 和 β 之间的专性异二聚体 其中ITGAV和ITGB5是已知的24个亚基之一。作为更重要的一对，ITGAV被探测 高密度 CRISPR 平铺扫描，我们发现一个小口袋对于 ITGAV 功能至关重要，它是 可修正小分子结合。基于结构的分析发现了 ITGAV β 对的环结构 与发现的口袋相互作用，导致我们假设该口袋对于异二聚体至关重要 ITGAV 及其 beta 对之间的稳定性。事实上，通过 500 个小分子的高通量筛选，我们 发现一种化合物似乎与我们的口袋结合并破坏ITGAV和ITGB5之间的异二聚体。 进一步验证这种潜力将是论文研究和后续工作要做的剩余工作 完成后，将填补未满足的临床需求，因为 FDA 尚未批准针对整合素的药物 用于癌症适应症。为了进一步提高治疗癌症的潜力，我计划使用数学模型 通过了解癌症信号传导的复杂性来确定新的治疗策略的方法 K00阶段，建议博士后工作。与博士合作研究复杂的癌症信号传导。 Pirrotte，我们在 ITGAV 被敲除的细胞中生成了激酶活性评分。利用这些数据我们可以建模 信号传导的影响，因为它与癌细胞中可测量的变化有关。具体来说，我们将研究细胞 循环控制，其因 ITGAV 丢失而受到抑制。此外，我们可以将已知的抑制剂建模为常见的 信号级联作为新型组合治疗策略。为了确认我们的模型，我将使用实验室 F99 阶段开发的基础技能。总的来说，通过 F99/K00 奖项的培训，我将获得以下技能： 能够建立数学模型来研究癌症并使用基于实验室的技能验证这些模型。 这将使我成为转化研究领域的独立研究和领先科学家。