Targeting a Novel Pocket on ITGAV

瞄准 ITGAV 上的新颖口袋

• 批准号: 10676274
• 负责人: NICOLE MATTSON
• 金额: $ 0.68万
• 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 2023-09-22
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10676274
• 关键词: Acute Lymphocytic Leukemia; Acute Myelocytic Leukemia; Advanced Malignant Neoplasm; Apoptosis; Award; Binding; Biological; Biological Assay; Biology; Breast; CRISPR screen; Cancer Model; Cause of Death; Cell Cycle; Cell Cycle Regulation; Cell Death Induction; 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; Invaded; Knock-out; Laboratories; Liquid substance; MAPK10 gene; Machine Learning; 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; Technology; Therapeutic; Time; Training; Translational Research; United States; Validation; Western Blotting; Work; beta catenin; biomarker discovery; cancer cell; cancer survival; cell killing; comparison control; 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; statistics; 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。在美国，癌症死亡人数仍然处于历史最高水平，这使得迫切的临床需求 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 目前的科学方法才刚刚开始揭示癌症的潜在复杂性。技术 进步正在迅速改变科学发现的景观;例如， 数学建模与基于实验室的验证相结合，导致更好的组合疗法， 治疗癌症因此，我建议与F99/K 00博士前博士后研究员进行培训 过渡奖。对于F99阶段，论文研究，我将专注于基于实验室的研究技能 来鉴定和提出一种治疗癌症的新疗法。在一个高水平的CRISPR筛选中， 通过对细胞表面基因的分析，我们发现整合素α V（ITGAV）对于实体瘤的存活是必需的 （结肠癌、胰腺癌和乳腺癌）。为了验证ITGAV作为最重要的整合素，我们设计了第二个 针对所有26种整联蛋白进行层筛选，发现ITGAV和整联蛋白β 5（ITGB 5）是唯一必需的 实体瘤中的整合素。有趣的是，整合素必须在α和β之间形成专性异二聚体， 其中ITGAV和ITGB 5是已知的24个亚基之一。作为更重要的一对，ITGAV用 高密度CRISPR平铺扫描，我们发现一个小口袋对ITGAV功能至关重要， 可与小分子结合。基于结构的分析发现ITGAV的β对的环结构 与发现的口袋相互作用，导致我们的假设，口袋是必不可少的异二聚体 ITGAV和它的β对之间的稳定性。事实上，从500个小分子的高通量筛选中， 发现了一种化合物，它似乎与我们的口袋结合，并破坏了ITGAV和ITGB 5之间的异二聚体。 进一步验证这一潜力将是剩余的工作要做的论文研究和 完成后，将填补未满足的临床需求，因为没有FDA批准的靶向整合素的药物 用于癌症适应症。为了进一步提高治疗癌症的潜力，我计划使用数学建模， 通过理解癌症信号传导的复杂性来确定新的治疗策略的方法， K 00阶段，拟从事博士后工作。为了研究复杂的癌症信号，与Dr。 Pirrotte，我们在ITGAV被敲除的细胞中生成激酶活性评分。有了这些数据， 信号的影响，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，其被ITGAV损失抑制。此外，我们可以将已知的抑制剂建模为常见的 信号级联作为新的组合治疗策略。为了确认我们的模型，我将使用实验室 在F99阶段开发的基础技能。总的来说，通过F99/K 00奖励的培训，我将获得以下技能： 能够建立数学模型来研究癌症，并利用实验室技能验证这些模型。 这将使我成为独立的研究和转化研究的领先科学家。