Effect of Cocaine and LTR Polymorphism on HIV-1 Pathogenesis

可卡因和 LTR 多态性对 HIV-1 发病机制的影响

• 批准号: 7691302
• 负责人: Richard B. Markham
• 金额: $ 57.56万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7691302
• 关键词: ATF2 gene; Acute; Address; Animal Model; Anti-Retroviral Agents; Antiretroviral resistance; Aquila; Binding; Binding Sites; Biological; Biological Models; Brain; CD4 Positive T Lymphocytes; Cells; Characteristics; Cocaine; Cocaine Abuse; Cocaine Users; Complex; Consensus Sequence; DNA Binding; Development; Dimerization; Disease Progression; Drug abuse; Drug usage; Drug user; Electrophoretic Mobility Shift Assay; Event; FOS gene; Family; Figs - dietary; Frequencies; Gene Components; Gene Expression; Genetic; Genetic Polymorphism; Genetic Transcription; Genetic Variation; HIV; HIV-1; Heterogeneity; Illicit Drugs; Immediate-Early Genes; In Vitro; Individual; Injection of therapeutic agent; JUN gene; Laboratory Study; Length; Leucine Zippers; Link; Long Terminal Repeats; Lymphocyte; Microglia; Molecular; Multivariate Analysis; Mutation; Outcome; Pathogenesis; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Population; Protease Inhibitor; Proteins; Recording of previous events; Regulation; Reporting; Resistance; Sampling; Specimen; Surface; System; Technology; Testing; Time; Transactivation; Transcription Factor AP-1; Variant; Viral; Viral Load result; Virus; Woman; addiction; antiretroviral therapy; base; brain cell; clinical care; cocaine exposure; cocaine use; cohort; cytokine; dimer; drug of abuse; in vivo; macrophage; medical schools; monocyte; non-drug; peripheral blood; promoter; public health relevance; resistance mutation; transcription factor; virus genetics

DESCRIPTION (provided by applicant): Previous studies from this laboratory have demonstrated significantly higher viral genetic diversity and a higher frequency of primary resistance to protease inhibitors (PI) among illicit drug users compared to HIV-1 infected non-drug users. To explore the potential link between drug use and greater viral replication, diversity and antiretroviral resistance, we propose to exploit the findings that 1) cocaine abuse stimulates high levels of the AP-1 transcription factor associated immediate early genes and 2) only about 40% of Clade B infected individuals carry an HIV-1 LTR length polymorphism (MFNLP) containing an AP-1 binding site. Using a large cohort of cocaine using HIV-1 infected subjects followed at the Vanderbilt Clinical Care Center, we will address the hypothesis that cocaine users infected with HIV-1 carrying the Clade B HIV-1 LTR polymorphism with an AP-1 binding site will have higher viral loads, greater genetic diversity, and greater primary resistance to antiretrovirals than either cocaine users whose virus does not carry the LTR polymorphism or non-cocaine users. To explore this hypothesis we will 1) Identify among a cohort of HIV-1 positive, antiretroviral therapy naive individuals those cocaine-using and non-cocaine using individuals with virus carrying the MFNLP within the LTR. We will thereby have identified four groups for additional analyses: HIV-1 infected cocaine users with and without the MFNLP and non-cocaine users with and without the MFNLP. 2) Within the four groups quantify the expression levels within peripheral blood CD4 T lymphocytes and monocytes of c-fos and JunB, as representative immediate early gene components of AP-1. Similarly, using gel-shift assays or a new high throughput kit, we will quantify levels of AP-1 within those cell populations from the four groups. 3) Using 454 high throughput sequencing technology we will sequence the protease region to define within host genetic diversity and primary resistance mutations to PI. Using diversity, resistance mutations to PI and viral load as outcomes, we will examine using a multivariate analysis, the effect of cocaine use, immediate early gene expression levels, AP-1 quantities, and the presence of the MFNLP. PUBLIC HEALTH RELEVANCE: Certain variants of HIV-1 carry genetic alterations that might enable them to replicate more rapidly in cells exposed to cocaine. This study will evaluate whether the presence of those variants in cocaine users alters various parameters of disease progression.

描述（由申请人提供）：该实验室之前的研究表明，与感染 HIV-1 的非吸毒者相比，非法吸毒者的病毒遗传多样性明显更高，对蛋白酶抑制剂 (PI) 的原发耐药率更高。为了探索吸毒与更大的病毒复制、多样性和抗逆转录病毒耐药性之间的潜在联系，我们建议利用以下发现：1) 可卡因滥用会刺激高水平的与立即早期基因相关的 AP-1 转录因子，2) 仅约 40% B 分支感染个体携带含有 AP-1 结合位点的 HIV-1 LTR 长度多态性 (MFNLP)。通过在范德比尔特临床护理中心对大量使用 HIV-1 感染受试者的可卡因进行跟踪，我们将解决这样的假设：感染携带带有 AP-1 结合位点的 Clade B HIV-1 LTR 多态性的 HIV-1 的可卡因使用者将与病毒不携带 LTR 多态性的可卡因使用者或非可卡因使用者相比，具有更高的病毒载量、更大的遗传多样性和更强的抗逆转录病毒药物初级耐药性。为了探索这一假设，我们将 1) 在一组 HIV-1 阳性、未接受过抗逆转录病毒治疗的个体中识别那些在 LTR 内携带 MFNLP 病毒的可卡因使用者和非可卡因使用者。因此，我们将确定四组进行额外分析：感染 HIV-1 的可卡因使用者（有或没有 MFNLP）和非可卡因使用者（有或没有 MFNLP）。 2)在四组中量化外周血CD4 T淋巴细胞和单核细胞中c-fos和JunB的表达水平，作为AP-1的代表性立即早期基因成分。同样，使用凝胶迁移测定或新的高通量试剂盒，我们将量化四组细胞群中 AP-1 的水平。 3) 使用454高通量测序技术，我们将对蛋白酶区域进行测序，以确定宿主遗传多样性和PI的主要抗性突变。使用多样性、PI 抗性突变和病毒载量作为结果，我们将使用多变量分析来检查可卡因使用的影响、早期基因表达水平、AP-1 数量以及 MFNLP 的存在。公共卫生相关性：HIV-1 的某些变体携带基因改变，可能使它们能够在接触可卡因的细胞中更快地复制。这项研究将评估可卡因使用者中这些变异的存在是否会改变疾病进展的各种参数。