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: 8263411
• 负责人: John S. Albin
• 金额: $ 5.12万
• 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/8263411
• 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; Health; Human; Infection; Lead; Mediating; Modeling; Mutate; Mutation; National Institute of Drug Abuse; Outcome; Patients; Peptide Sequence Determination; Pharmaceutical Preparations; Protein Family; Proteins; Proviruses; Regimen; Research; Resistance; Resistance development; Reverse Transcription; Role; Testing; Treatment Failure; Viral; Virion; Virus; apolipoprotein B mRNA editing enzyme; base; design; improved; killings; polypeptide; programs; 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.

性状（由申请方提供）：人APOBEC 3蛋白通过逆转录过程中病毒cDNA的突变抑制人免疫缺陷病毒1型（HIV-1）的复制。在足够数量的情况下，这些突变导致前病毒失活。在患者中，APOBECS介导的限制不能控制病毒复制，因为最有效的APOBEC 3蛋白被病毒辅助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APOBEC 3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抗性的HIV-1点突变发生在APOBEC 3突变热点，这增加了病毒实际上颠覆APOBEC 3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APOBEC 3蛋白引入的亚致死水平的突变有助于获得HIV-1耐药性。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，通过避免使用易受APOBECS介导的耐药性影响的药物来最大限度地减少治疗失败。这些战略有可能极大地改善对药物滥用使治疗复杂化的人的护理，并与国家药物滥用研究所艾滋病研究方案2008年供资优先事项的“重点领域3：药物使用与艾滋病毒/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。具体目标1 -定义能够突变Vif-proficient HIV-1的APOBEC 3蛋白：这些实验将定量确定哪些APOBEC 3蛋白存在于活化的CD 4 + T细胞中，以及这些蛋白在感染早期受到抑制的程度。进一步的实验将定量Vif-proficient病毒体中APOBEC 3蛋白的酸化和突变活性。具体目标2 -确定APOBEC 3蛋白是否影响耐药性：该目标集中于HIV-1在APOBEC 3蛋白存在下的生长是否影响感染细胞培养模型中耐药性的获得。进一步的实验将确定APOBEC 3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突变从而对这些药物产生耐药性的能力的影响。这些实验将确定被称为APOBEC 3的天然人类蛋白质是否能增强HIV对用于治疗HIV感染的药物的耐药性。这将对优化抗HIV治疗产生影响。