Determining the Role of APOBEC3 Proteins in HIV-1 Drug Resistance

确定 APOBEC3 蛋白在 HIV-1 耐药性中的作用

• 批准号: 7623283
• 负责人: John S. Albin
• 金额: $ 3.92万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-25 至 2013-05-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7623283
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Affect; Amino Acid Sequence; Anti-HIV Therapy; Anti-Retroviral Agents; Antiretroviral drug resistance; Antiviral Agents; Area; CD4 Positive T Lymphocytes; Caring; Cell Culture Techniques; Cells; Complementary DNA; Cytosine deaminase; DNA; Drug Addiction; Drug abuse; Drug resistance; Drug usage; Funding; Growth; HIV; HIV Infections; HIV-1; Human; Infection; Lead; Mediating; Modeling; Mutate; Mutation; National Institute of Drug Abuse; Outcome; Patients; Peptide Sequence Determination; Pharmaceutical Preparations; Protein Family; Proteins; Proviruses; Research; Resistance; Resistance development; Reverse Transcription; Role; T Virus; Testing; Treatment Failure; Treatment Protocols; Viral; Virion; Virus; apolipoprotein B mRNA editing enzyme; base; design; human apolipoprotein B mRNA editing enzyme; improved; killings; polypeptide; programs; public health relevance; research study; success

DESCRIPTION (provided by applicant): Human APOBEC3 proteins inhibit the replication of human immunodeficiency virus type 1 (HIV-1) via the mutation of viral cDNA during reverse transcription. In sufficient numbers, these mutations lead to the inactivation of proviruses. In patients, APOBECS-mediated restriction is incapable of controlling viral replication because the most potent APOBEC3 proteins are neutralized by the viral accessory protein virion infectivity factor (Vif). Nevertheless, there may be an intermediate scenario in which some APOBEC3 proteins escape degradation and mutate HIV-1 without affecting viral growth. Interestingly, many HIV-1point mutations conferring resistance to antiretroviral drugs occur inAPOBEC3 mutational hotspots, raising the possibility that the virus actually subverts APOBEC3-mediated mutation to enhance its ability to resist treatment. Thus, we hypothesize that sublethal levels of mutation introduced by APOBEC3 proteins contribute to the acquisition of HIV-1 drug resistance. If proven, this will provide a basis for the rational design of antiretroviral regimens that minimize treatment failure by avoiding drugs susceptible to APOBECS-mediated resistance. Such strategies carry the potential to greatly improve the care of people for whom drug abuse complicates treatment and are consistent with "Area of Emphasis 3: Drug Use and HIV/AIDS" of the 2008 funding priorities of the AIDS Research Program at the National Institute on Drug Abuse. To test our hypothesis, we propose two specific aims. Specific Aim 1 - Define the APOBEC3 Proteins Capable of Mutating Vif-proficient HIV-1: These experiments will quantitatively determine which APOBEC3 proteins are present in activated CD4+ T cells and to what extent these proteins are suppressed early in infection. Further experiments will quantify the encapsidation and mutational activity of APOBEC3 proteins in Vif-proficient virions. Specific Aim 2 - Determine Whether APOBEC3 Proteins Impact Drug Resistance: This aim focuses on whether growth of HIV-1in the presence of APOBEC3 proteins affects the acquisition of drug resistance in a cell culture model of infection. Additional experiments will determine whether the depletion of APOBEC3 proteins decreases the ability of HIV-1to develop resistance. PUBLIC HEALTH RELEVANCE: The success of antiviral drugs is tempered by the ability of viruses to acquire mutations conferring resistance to those drugs. These experiments will determine whether natural human proteins known as APOBEC3 enhance the ability of HIV to evolve resistance to the medications used to treat HIV infection. This will have implications for the optimization of anti-HIV therapy.

描述（由申请人提供）：人类APOBEC3蛋白在逆转录过程中通过病毒cDNA突变抑制人类免疫缺陷病毒1型（HIV-1）的复制。在足够数量的情况下，这些突变导致原病毒失活。在患者中，apobecs介导的限制不能控制病毒复制，因为最有效的APOBEC3蛋白被病毒附属蛋白病毒粒子感染因子（Vif）中和。然而，可能存在一种中间情况，其中一些APOBEC3蛋白逃避降解并使HIV-1突变而不影响病毒生长。有趣的是，许多赋予抗逆转录病毒药物耐药性的hiv -1点突变发生在apobec3突变热点，这增加了病毒实际上破坏apobec3介导的突变以增强其抵抗治疗能力的可能性。因此，我们假设由APOBEC3蛋白引入的亚致死水平的突变有助于HIV-1耐药性的获得。如果得到证实，这将为合理设计抗逆转录病毒治疗方案提供基础，通过避免易受apobecs介导的耐药性影响的药物，最大限度地减少治疗失败。这种策略有可能极大地改善对吸毒使治疗复杂化的人的护理，并且与国家药物滥用研究所艾滋病研究计划2008年资助优先事项中的“重点领域3：吸毒和艾滋病毒/艾滋病”相一致。为了验证我们的假设，我们提出了两个具体目标。特异性目标1 -确定APOBEC3蛋白能够突变精通vif的HIV-1：这些实验将定量确定哪些APOBEC3蛋白存在于活化的CD4+ T细胞中，以及这些蛋白在感染早期被抑制的程度。进一步的实验将量化APOBEC3蛋白在vif精通病毒粒子中的衣壳化和突变活性。特异性目标2 -确定APOBEC3蛋白是否影响耐药性：该目标关注在感染细胞培养模型中，APOBEC3蛋白存在下hiv -1的生长是否影响耐药性的获得。进一步的实验将确定APOBEC3蛋白的缺失是否会降低hiv -1产生耐药性的能力。公共卫生相关性：抗病毒药物的成功受到病毒获得对这些药物产生耐药性的突变能力的影响。这些实验将确定被称为APOBEC3的天然人类蛋白是否会增强HIV进化出对用于治疗HIV感染的药物的耐药性的能力。这将对优化抗hiv治疗产生影响。