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: 7849075
• 负责人: Richard B. Markham
• 金额: $ 56.9万
• 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/7849075
• 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; DNA Binding; Development; Dimerization; Disease Progression; Drug abuse; Drug usage; Drug user; Electrophoretic Mobility Shift Assay; Event; FOS gene; Family; Frequencies; Gene Components; Gene Expression; Genetic; Genetic Polymorphism; Genetic Transcription; Genetic Variation; HIV-1; Heterogeneity; Homo; 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）：与HIV-1感染的非药物使用者相比，该实验室的先前研究表明，在非法吸毒者中，非法药物使用者对蛋白酶抑制剂（PI）的一级抗药性频率更高，并且对蛋白酶抑制剂（PI）的频率更高。为了探索药物使用与更大的病毒复制，多样性和抗逆转录病毒抗性之间的潜在联系，我们建议利用以下发现，即1）可卡因滥用刺激高水平的AP-1转录因子与早期基因相关的高水平的AP-1转录因子，而2）仅约40％受体B感染的个体携带含有AP-1结合位点的HIV-1 LTR长度多态性（MFNLP）。使用大量可卡因使用HIV-1感染的受试者随后在范德比尔特临床护理中心进行，我们将解决以下假设：可卡因使用者感染了带有AP-1结合部位的HIV-1的可卡因用户，携带甲状管甲基-1 LIV-1 LTR多态性将会与可卡因使用者相比，病毒使用者的病毒量更高，遗传多样性更大，对抗逆转录病毒的主要抵抗力更大，可卡因使用者不会携带LTR多态性或非cocaine使用者。为了探讨这一假设，我们将在HIV-1阳性，抗逆转录病毒疗法的队列中确定那些使用可卡因的人，并使用携带MFNLP在LTR中的病毒的个体来识别那些使用可卡因和非卡因。因此，我们将确定四个组以进行其他分析：HIV-1感染了有或没有MFNLP的MFNLP和非卡通用户的可卡因用户。 2）在四个组中，量化了C-FOS和JUNB的外周血CD4 T淋巴细胞和单核细胞中的表达水平，为AP-1的代表性早期基因成分。同样，使用凝胶转移测定或新的高吞吐量试剂盒，我们将在四组中量化这些细胞群体中的AP-1水平。 3）使用454个高通量测序技术，我们将对蛋白酶区域进行测序，以定义宿主遗传多样性和对PI的一级抗性突变。使用多样性，对PI和病毒负荷作为结果的抗性突变，我们将使用多变量分析，可卡因使用的影响，立即的早期基因表达水平，AP-1量以及MFNLP的存在进行检查。公共卫生相关性：HIV-1的某些变体携带遗传改变，可能使它们能够在暴露于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体的存在是否会改变疾病进展的各种参数。