Decoding Viral Control of Host Kinase Signaling to Design Combination Therapy

解码病毒对宿主激酶信号传导的控制以设计联合疗法

• 批准号: 10449933
• 负责人: Mehdi Bouhaddou
• 金额: $ 12.5万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-12 至 2023-02-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10449933
• 关键词: 2019-nCoV; Advisory Committees; African Green Monkey; Antiviral Agents; Antiviral Therapy; Area; Biochemical; Bioinformatics; Biological Assay; Biology; COVID-19; COVID-19 treatment; CRISPR interference; Cell Culture Techniques; Cell Survival; Cell physiology; Cells; Clinical Trials; Collecting Cell; Combined Modality Therapy; Common Cold; Communicable Diseases; Coronavirus; Coronavirus Infections; Cyclin-Dependent Kinases; Data; Data Set; Dependence; Development; Disease; Drug Compounding; Drug Targeting; Economics; Family; Genetic; Genetic Screening; Global Change; Health Hazards; Hour; Human; Human Cell Line; Immune response; Infection; Infectious Agent; Interruption; Investigational Therapies; Knowledge; Life Cycle Stages; Light; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Mediator of activation protein; Mentors; Methods; Mission; Modeling; Pathway Analysis; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Phenotype; Phosphorylation; Phosphotransferases; Post-Translational Protein Processing; Process; Production; Proteins; Proteomics; Public Health; RNA; Regulation; Research; Research Personnel; Research Priority; Resistance; Resistance development; Respiratory Disease; SARS-CoV-2 infection; Signal Transduction; Testing; Therapeutic; Time; Training; Translating; United States National Institutes of Health; Vaccines; Viral; Viral Proteins; Virion; Virus; Virus Replication; Work; anti-viral efficacy; base; betacoronavirus; bioinformatics tool; casein kinase II; cell type; combat; combinatorial; coronavirus disease; curative treatments; design; druggable target; fitness; global health; insight; mathematical model; network models; novel; novel therapeutic intervention; p38 Mitogen Activated Protein Kinase; pandemic coronavirus; pandemic disease; pathogen; phosphoproteomics; public health emergency; skills; theories; therapeutic development; transcriptome sequencing; treatment strategy; virology

PROJECT SUMMARY Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a single-stranded RNA beta-coronavirus of the family Coronaviridae that emerged in late 2019 and causes the infectious disease COVID-19 (Coronavirus Disease-2019). The emergence of the global pandemic has resulted in a public health emergency with detrimental societal and global economic effects. At present, there are no vaccines or curative treatments available. The search for antiviral therapies constitutes a global research priority. Combinatorial host-directed antiviral treatments present a low propensity for resistance and a high potential for potent efficacy. Pathogens often provoke dramatic changes to host cellular signaling to enhance their fitness. Proteomic approaches that globally quantify changes in protein abundance and phosphorylation represent an unbiased way to elucidate how host pathways are rewired during infection. We previously observed that SARS-CoV-2 provokes dramatic changes to host phosphorylation, suggesting the regulation of host kinases to be an important mediator of coronavirus infection and disease. We additionally found many SARS-CoV-2 proteins to physically interact with several human kinases. Furthermore, we and others have shown that targeting host kinases hijacked by SARS-CoV-2 is an effective means of discovering antiviral therapies. We identified 49 kinases to be dysregulated during infection—including p38, casein kinase II, cyclin dependent kinases, AXL, and PIKFYVE— finding 38/68 (>50%) drugs targeting these kinases to be antiviral in cell culture assays (CC50/IC50>10). Here, I propose to expand upon my prior work by (Aim 1) mapping the global regulation of host phosphorylation signaling by SARS-CoV-2 and OC43, a non-lethal coronavirus and cause of common cold, infection in human cells using mass spectrometry-based proteomics to identify novel kinases and pathways dysregulated during coronavirus infection. Next, I will (Aim 2) validate coronavirus dependency on host responses and use this information to design and test novel combination therapy to treat infection in human cell culture models. Lastly, I will (Aim 3) decode how kinases impinge upon the coronavirus life cycle as well as how kinases control phosphorylation of viral/host proteins during infection. This systematic approach will shed light on coronavirus biology and therapeutic strategies, with general applicability to other infectious agents and diseases. My request for additional mentored training is based on a need to master skills in (1) mass spectrometry-based proteomics, (2) virology theory and methods, and (3) functional genetics. My background is in mathematical modeling, bioinformatics, and cancer, which I plan to integrate with skills in proteomics, genetics, and virology to gain independence from my advisor and start a dual wet/dry independent research lab studying post-translational modifications and signaling rewiring in infectious disease and cancer. My mentor, co-mentor, and advisory committee will train and mentor me as I progress towards becoming an independent investigator.

项目摘要 严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是一种单链RNA β-冠状病毒， 2019年底出现的冠状病毒科，导致传染病COVID-19（冠状病毒 疾病（2019年）。全球大流行的出现导致了公共卫生紧急情况， 有害的社会和全球经济影响。目前还没有疫苗或治愈性治疗方法 available.寻找抗病毒疗法是全球研究的优先事项。组合主机导向 抗病毒治疗呈现出低的耐药性倾向和高的强效潜力。 病原体通常会引起宿主细胞信号的剧烈变化，以增强其适应性。蛋白组 全球量化蛋白质丰度和磷酸化变化的方法代表了一种无偏的 阐明宿主在感染过程中如何重新连接途径的方法。我们之前观察到SARS-CoV-2 引起宿主磷酸化的显著变化，表明宿主激酶的调节是一个重要的 冠状病毒感染和疾病的介质。我们还发现了许多SARS-CoV-2蛋白， 与几种人类激酶相互作用。此外，我们和其他人已经表明，靶向宿主激酶， 被SARS-CoV-2劫持是发现抗病毒疗法的有效手段。我们鉴定了49种激酶， 感染期间失调-包括p38、酪蛋白激酶II、细胞周期蛋白依赖性激酶、AXL和PIKFYVE- 在细胞培养试验中发现38/68（>50%）靶向这些激酶的药物是抗病毒的（CC 50/IC 50>10）。 在这里，我建议通过（目标1）绘制宿主磷酸化的全局调节来扩展我先前的工作 SARS-CoV-2和OC 43（一种非致死性冠状病毒和普通感冒的原因）在人类感染中的信号传导 细胞使用基于质谱的蛋白质组学来识别新的激酶和途径， 冠状病毒感染。接下来，我将（目标2）验证冠状病毒对宿主反应的依赖性，并使用 设计和测试新的联合疗法以治疗人类细胞培养模型中的感染。最后， 我将（目标3）解码激酶如何影响冠状病毒的生命周期，以及激酶如何控制 在感染期间病毒/宿主蛋白的磷酸化。这种系统的方法将揭示冠状病毒 生物学和治疗策略，一般适用于其他传染源和疾病。 我要求额外的指导培训是基于需要掌握以下技能：（1）基于质谱的 蛋白质组学，（2）病毒学理论和方法，（3）功能遗传学。我的背景是数学 建模，生物信息学和癌症，我计划将其与蛋白质组学，遗传学和病毒学的技能相结合， 从我的导师那里获得独立，并开始了一个双湿/干独立研究实验室，研究翻译后 感染性疾病和癌症中的修饰和信号重连。我的导师，共同导师，和顾问 委员会将在我成为独立调查员的过程中对我进行培训和指导。