A Method to Stop HIV Replication:Inhibition of Human Purine Utilizing Proteins

阻止HIV复制的方法：抑制人嘌呤利用蛋白

基本信息

批准号：
8075457
负责人：
JESSE J KWIEK
金额：
$ 50.26万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-01 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8075457
关键词：
Affinity Amino Acids Antineoplastic Agents Antiviral Agents Attenuated Bacterial Infections Binding Binding Proteins Biology CD4 Positive T Lymphocytes Cell Culture Techniques Cells Clinical Trials Communicable Diseases Coupled DNA Databases Development Down-Regulation Drug Delivery Systems Drug resistance Enzymes Erectile dysfunction Event Funding Generations Generic Drugs Goals HIV HIV Infections HIV drug resistance HIV-1 Heat shock proteins Human Hypertension Infection Jurkat Cells Lead Life Cycle Stages Link Malignant Neoplasms Mass Spectrum Analysis Methods Modeling Monomeric GTP-Binding Proteins NADH Non-Insulin-Dependent Diabetes Mellitus Online Mendelian Inheritance In Man Oxidoreductase Pathway interactions Pharmaceutical Preparations Phase Plant Resins Predisposition Process Protein Binding Protein Kinase Proteins Proteome Proteomics Purines RNA chemical synthesis Regulation Research Resistance development Retroviridae Reverse Transcription Role Small Interfering RNA Stable Isotope Labeling System TRIM Gene Technology Therapeutic Time Transcription Process Vaccines Validation Vertebral column Viral Viral Proteins Virus Wolves Work base conventional therapy design drug discovery experience functional genomics helicase human disease hypercholesterolemia innovation innovative technologies macrophage microbicide monocyte novel novel strategies pandemic disease protein activation protein expression protein folding purine small molecule therapeutic target tissue culture tissue/cell culture

项目摘要

DESCRIPTION (provided by applicant): Owing to the recent setbacks in vaccine and microbicide trails, novel strategies to slow the HIV-pandemic are desperately needed. Development of a small molecule that blocked a host process required for HIV-1 replication could provide such a strategy. Like all retroviruses, HIV-1 requires host proteins to complete its life cycle, and these intracellular host molecules represent an undeveloped pool of novel anti-HIV therapeutics. The goal of this application is to use functional proteomics to identify host proteins that are dispensable to the host but essential for viral replication. A generic proteomic screen would likely identify numerous host proteins induced by HIV-infection, most of which would be poor therapeutic candidates. A functional proteomics screen, which queries a subset of proteins with strong therapeutic potential, would greatly simplify the search for host targets; the host purinome has this potential. The purinome comprises any protein that binds purine- containing molecules (e.g. ATP, NADH), and it includes heat shock proteins, dehydrogenases, and protein kinases. Inhibition of purinome proteins forms the basis of many current therapies, including those targeting cancer, hypertension, and bacterial infections. The central hypothesis of this application states that inhibition of purinome proteins, which are induced or regulated by HIV-infection, will block HIV-1 replication. Our research team has designed a protein affinity media that captures the entire purinome through its purine-binding pocket. The proposed research will proceed in four broad steps: use SILAC (stable isotope labeling with amino acids in cell culture) to quantify purinome proteins induced by HIV-1 infection of THP1 and Jurkat cells (target identification); determine which purinome proteins are essential for HIV-1 replication (target validation); identify drugs that selectively compete validated targets from the purinome-binding resin (drug discovery); confirm that these compounds block HIV-replication in primary cells (drug validation). During this process, we will also identify host proteins that interact with HIV-1, and in the event that a viable purinome target and/or drug is not identified, viral biology will be advanced by a greater understanding of how HIV-1 interacts with its host. The proposed research uses innovative proteomics technology (SILAC) and an unconventional, yet validated, approach. First, conventional anti-HIV therapeutics inhibit virally-encoded proteins, which makes them more susceptible to the development of resistance. By targeting a host enzyme that is required for HIV-1 replication, it is likely that the virus will have to undergo more radical changes to offset the changes in the host milieu. Second, conventional drug discovery uses a reductionist approach to separately identify a target and a drug, while our approach combines the target/drug identification process, which makes discovery more efficient. At the completion of this project, our combination of experience and technology will enable us to characterize host purine-binding proteins that are essential for HIV-1 replication and identify several lead compounds that will inhibit host proteins necessary for HIV-1 replication.

描述（由申请人提供）：由于最近在疫苗和杀生型踪迹中遇到的挫折，迫切需要减慢艾滋病毒大流行的新型策略。开发封闭HIV-1复制所需的宿主过程的小分子可以提供这样的策略。像所有逆转录病毒一样，HIV-1需要宿主蛋白来完成其生命周期，这些细胞内宿主分子代表了未开发的新型抗HIV疗法池。该应用的目的是使用功能蛋白质组学来识别宿主可分配但对于病毒复制至关重要的宿主蛋白。通用的蛋白质组学筛查可能会鉴定出由HIV感染引起的许多宿主蛋白，其中大多数是治疗性较差的候选者。一个功能性蛋白质组学筛选，它查询具有强大治疗潜力的蛋白质的子集，将极大地简化了对宿主目标的搜索。宿主嘌呤组具有这种潜力。尖嘌呤组包括任何结合含嘌呤的分子（例如ATP，NADH）的蛋白质，并包括热休克蛋白，脱氢酶和蛋白激酶。嘌呤组蛋白的抑制是许多当前疗法的基础，包括针对癌症，高血压和细菌感染的疗法。该应用的中心假设指出，受HIV感染诱导或调节的尖嘌呤蛋白的抑制作用将阻止HIV-1复制。我们的研究团队设计了一种蛋白质亲和力媒体，该媒体通过其嘌呤结合口袋捕获了整个Purinome。拟议的研究将以四个广泛的步骤进行：使用SILAC（在细胞培养中使用氨基酸稳定的同位素标记）来量化由THP1和Jurkat细胞HIV-1感染诱导的嘌呤组蛋白（靶标识别）；确定哪种嘌呤组蛋白对于HIV-1复制至关重要（目标验证）；确定从果嘌呤结合树脂（药物发现）中有选择性竞争验证靶标的验证靶标的药物；确认这些化合物阻止了原代细胞中的HIV复制（药物验证）。在此过程中，我们还将确定与HIV-1相互作用的宿主蛋白，如果未鉴定出可行的嘌呤组靶标和/或药物，则通过对HIV-1如何与HIV与HIV相互作用的方式进行更深入的了解，病毒生物学将得到推进。它的主人。拟议的研究使用创新的蛋白质组学技术（SILAC）和非常规但经过验证的方法。首先，常规的抗HIV疗法抑制了病毒编码的蛋白质，这使它们更容易受到抗性的发展。通过瞄准HIV-1复制所需的宿主酶，该病毒可能必须进行更多的根本变化以抵消宿主环境中的变化。其次，常规药物发现采用还原论的方法来单独识别靶标和药物，而我们的方法结合了目标/药物识别过程，这使发现更加有效。该项目完成后，我们的经验和技术组合将使我们能够表征宿主的嘌呤结合蛋白，这些蛋白质对于HIV-1复制至关重要，并识别几种将抑制HIV-1复制所需的宿主蛋白的铅化合物。