Systematic Targeting of Oncogene Interacting Proteins to Reveal New Therapeutic Strategies

系统靶向癌基因相互作用蛋白以揭示新的治疗策略

基本信息

批准号：
9756188
负责人：
Mehdi Bouhaddou
金额：
$ 6.09万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9756188
关键词：
Adaptor Signaling Protein Aerobic Affinity Chromatography Binding Biochemical Biological Process CRISPR/Cas technology Cancer cell line Cell Proliferation Cell physiology Cells Cessation of life Clustered Regularly Interspaced Short Palindromic Repeats Complementary DNA Complex Computing Methodologies Data Development Differential Equation Disease Enzyme-Linked Immunosorbent Assay FRAP1 gene GAB1 gene Genes Genetic Glucose Transporter Goals Growth Head and Neck Cancer Human Hyperglycemia Interruption Knock-out Knowledge Link Lipids Malignant - descriptor Malignant Neoplasms Mass Spectrum Analysis Measurement Mediating Mediator of activation protein Metabolic Metabolism Mission Modeling Mutate Oncogenes Oncogenic Outcome PIK3CA gene Pathway interactions Pharmacology Phenotype Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols Phosphotransferases Problem Solving Proteins Proteomics Proto-Oncogene Proteins c-akt Public Health Regulation Research Research Proposals Role Signal Pathway Signal Transduction Specificity Techniques Technology Testing Therapeutic Toxic effect United States National Institutes of Health Work base blood glucose regulation cancer initiation cancer therapy cell growth cell growth regulation cell motility clinical development computer framework design experimental study fighting fitness insight kinase inhibitor knockout gene live cell microscopy mathematical model mutant network models new therapeutic target novel novel therapeutic intervention novel therapeutics overexpression phosphatidylinositol 3,4,5-triphosphate phosphoproteomics programs protein protein interaction recruit side effect systemic toxicity therapeutic target tumor progression

项目摘要

PROJECT SUMMARY/ABSTRACT Cancer is an intimate part of us, recruiting a complex array of endogenous cellular processes to drive its fitness. Cancer initiation and progression is frequently the result of coordinated dysregulation of multiple signaling pathways by key oncogenes that strategically coopt the cancer signaling network. These oncogenes rarely work in isolation but instead form intricate higher-order complexes and participate in multifaceted networks of protein- protein interactions. One of the most commonly mutated proteins across all of human cancers is the phosphoinositide 3-kinase (PI3K) oncogene, a lipid kinase that can exploit diverse cellular programs to drive disease, including increased proliferation, survival, motility, cell growth and metabolic activity. Broad inhibition of PI3K is known to generate systemic toxicities, especially metabolic, which limit its clinical development. However, targeting PI3K interacting proteins may enable a safer alternative, by interrupting PI3K oncogenic activity while minimizing metabolic dysregulation. The long-term goal of this proposal is to deepen and refine our understanding of oncogene regulation of cancer signaling networks using systematic genetic, proteomic, and mathematical modeling approaches. The overall objective of this proposal is to identify the role of PI3K interacting proteins in modulating PI3K activity and recruitment of downstream cellular processes and to use this understanding to identify alternative therapeutic targets. This objective will be reached by testing the central hypothesis that PI3K interacting proteins, or downstream signaling pathways, can be modulated to tune PI3K activity and specificity, with the potential to simultaneously reduce malignancy and systemic toxicity. To test this hypothesis, the following three aims will be pursued. (Aim 1) Reveal Regulation of PI3K Activity and Cancer Phenotypes by PIK3CA Interacting Proteins. This aim will use CRISPR/Cas9 gene knockout technology to systematically delete and overexpress genes corresponding to PIK3CA interacting proteins and use live cell microscopy, combined with biochemical measurements, to assess the resulting impact on cell proliferation, survival, growth, motility, and metabolism. (Aim 2) Elucidate Proteomic Exploitation by PI3K as Mediated by PIK3CA Interacting Proteins. Here, PIK3CA interactors that preferentially bind the common H1047R mutant, as well as hits identified from Aim 1, will be subject to global proteomics and phosphoproteomics profiling to identify signaling pathways and biological processes regulated by each PIK3CA interacting protein. (Aim 3) Delineate Mechanisms of PI3K-mediated Manipulation of Pro-Cancer Signaling. Here, a novel computational framework will be developed by uniting data-driven network propagation techniques with mechanistic ordinary differential equation (ODE) modeling to delineate mechanistic pathways linking each PIK3CA interacting protein to its downstream effect. Successful completion of the proposed research will greatly enhance our mechanistic understanding of oncogene regulation in cancer. This will be a significant contribution as it will reveal novel therapeutic strategies to fight cancer while minimizing systemic toxicities.

项目概要/摘要癌症是我们亲密的一部分，它招募一系列复杂的内源性细胞过程来驱动其健康。癌症的发生和进展通常是多种信号传导协调失调的结果战略性地利用癌症信号网络的关键癌基因的通路。这些致癌基因很少起作用孤立地，而是形成复杂的高阶复合物并参与多方面的蛋白质网络蛋白质相互作用。所有人类癌症中最常见的突变蛋白之一是磷酸肌醇 3 激酶 (PI3K) 癌基因，一种脂质激酶，可以利用多种细胞程序来驱动疾病，包括增殖、存活、运动、细胞生长和代谢活动的增加。广泛抑制已知 PI3K 会产生全身毒性，尤其是代谢毒性，这限制了其临床开发。然而，靶向 PI3K 相互作用蛋白可以通过中断 PI3K 致癌活性来实现更安全的替代方案最大限度地减少代谢失调。该提案的长期目标是深化和完善我们的利用系统遗传学、蛋白质组学和方法了解癌症信号网络的癌基因调控数学建模方法。该提案的总体目标是确定 PI3K 相互作用的作用调节 PI3K 活性和招募下游细胞过程的蛋白质并使用它了解以确定替代治疗靶点。这一目标将通过测试中央假设可以通过调节 PI3K 相互作用蛋白或下游信号通路来调整 PI3K 活性和特异性，有可能同时减少恶性肿瘤和全身毒性。为了测试这个假设，将追求以下三个目标。（目标 1）揭示 PI3K 活性与癌症的调控 PIK3CA 相互作用蛋白的表型。该目标将利用CRISPR/Cas9基因敲除技术系统地删除和过度表达与 PIK3CA 相互作用蛋白相对应的基因并使用活细胞显微镜结合生化测量，评估对细胞增殖的影响，生存、生长、运动和新陈代谢。（目标 2）阐明 PI3K 介导的蛋白质组开发 PIK3CA 相互作用蛋白。此处，PIK3CA 相互作用子优先结合常见的 H1047R 突变体，如以及从目标 1 中识别出的命中，将接受全局蛋白质组学和磷酸化蛋白质组学分析，以识别每个 PIK3CA 相互作用蛋白调节的信号通路和生物过程。（目标 3）描绘 PI3K 介导的促癌信号转导操纵机制。这里，一个新颖的计算框架将通过将数据驱动的网络传播技术与机械常微分相结合来开发方程 (ODE) 建模，描绘将每个 PIK3CA 相互作用蛋白与其自身连接的机制途径下游效应。成功完成拟议的研究将大大增强我们的机制了解癌症中癌基因的调控。这将是一个重大贡献，因为它将揭示新颖的抗癌的治疗策略，同时最大限度地减少全身毒性。