High Specificity HIV-1 Markers Predictive of Neuro-AIDS

预测神经艾滋病的高特异性 HIV-1 标记物

• 批准号: 7119495
• 负责人: Brian Wigdahl
• 金额: $ 36.62万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7119495
• 关键词: AIDS dementia complex; HIV infections; biotechnology; clinical research; genetic markers; genetic registry /resource /referral center; genetic techniques; human fetus tissue; human immunodeficiency virus 1; human subject; molecular biology information system; nucleic acid repetitive sequence; single nucleotide polymorphism; technology /technique development; virus DNA; virus genetics

DESCRIPTION (provided by applicant): The progression of human immunodeficiency virus type 1 (HIV-1 )-associated disease in the immune and central nervous systems is associated with the ability of the virus to localize and replicate in specific cell populations within these compartments. Recent studies have suggested the possibility that genetic alterations within the HIV-1 genome introduced during viral replication may be correlated with either the stage of HIV-1 disease and/or neurologic status. We have recently demonstrated that a viral transactivator protein, Vpr, exhibits enhanced affinity for specific HIV-1 LTR C/EBP binding site configurations that can result in enhanced long terminal repeat (LTR) activation and correlate with end stage disease and HIVD. Additional studies have led to the identification of a number of HIV-1 LTR sequence signatures that increase in frequency within the peripheral blood compartment during progressive disease and track with the development of HIVD. These observations form the basis for the proposed investigations. The working hypothesis of these studies is that C/EBP and Sp binding site signatures within the LTR, which are selected for during viral replication over the course of HIV-1 disease, can be used as molecular markers to identify HIV-1-infected individuals that may be more prone to develop HIVD. The specific aims of this application are to (1) establish an HIV-1 LTR clone bank and sequence database derived from HIV-1-infected immune cell populations in the peripheral blood collected longitudinally from patients with defined clinical histories with respect to anti-retroviral therapy, drugs of abuse, and neurologic status, and from CNS-resident cells (microglial cells, perivascular macrophages, and astrocytes) at the time of autopsy; (2) analyze the LTR sequence database for the presence of the 3T C/EBP site I and 5TSp site III markers (and potential markers in other LTR regions), and establish correlations between marker prevalence and disease progression and severity, neurologic status, anti-retroviral therapy, and use of drugs of abuse; (3) develop a single nucleotide polymorphism (SNP) genotype assay for the detection of the 3T C/EBP site I and 5T Sp site III marker (and potential markers in other regions of the LTR) for use as a diagnostic assay indicative and/or predictive of peripheral disease progression and neurologic status; and (4) determine the ability of LTR clones containing the viral marker(s) to support transient expression in cell types of immune, neuroglial and bone marrow origin, and establish correlations between LTR function and clinical parameters established in previous aims. These studies may lead to the identification of additional tools to predict the development of HIVD and, in turn, provide more information to guide the therapeutic management of the HIV-1-infected patient.

描述（由申请人提供）：免疫和中枢神经系统中人类免疫缺陷病毒 1 型（HIV-1）相关疾病的进展与病毒在这些区室内的特定细胞群中定位和复制的能力有关。最近的研究表明，病毒复制过程中引入的 HIV-1 基因组内的遗传改变可能与 HIV-1 疾病的阶段和/或神经系统状态相关。我们最近证明，病毒反式激活蛋白 Vpr 对特定 HIV-1 LTR C/EBP 结合位点构型表现出增强的亲和力，可导致长末端重复 (LTR) 激活增强，并与终末期疾病和 HIVD 相关。其他研究已鉴定出许多 HIV-1 LTR 序列特征，这些特征在疾病进展期间外周血区室内的频率增加，并随着 HIVD 的发展而变化。这些观察结果构成了拟议调查的基础。这些研究的工作假设是，LTR 内的 C/EBP 和 Sp 结合位点特征（在 HIV-1 疾病过程中的病毒复制过程中被选择）可用作分子标记来识别 HIV-1 感染者可能更容易患艾滋病毒。本申请的具体目标是 (1) 建立 HIV-1 LTR 克隆库和序列数据库，这些克隆库和序列数据库源自从具有抗逆转录病毒治疗明确临床史的患者纵向收集的外周血中感染 HIV-1 的免疫细胞群。治疗、滥用药物和神经系统状态，以及尸检时中枢神经系统驻留细胞（小胶质细胞、血管周围巨噬细胞和星形胶质细胞）； (2) 分析 LTR 序列数据库中是否存在 3TC/EBP 位点 I 和 5TSp 位点 III 标记（以及其他 LTR 区域中的潜在标记），并建立标记流行率与疾病进展和严重程度、神经系统状态、抗-逆转录病毒疗法和滥用药物的使用； (3) 开发单核苷酸多态性 (SNP) 基因型测定，用于检测 3T C/EBP 位点 I 和 5T Sp 位点 III 标记（以及 LTR 其他区域的潜在标记），用作诊断测定指示和/或预测周围疾病进展和神经系统状态； (4)确定含有病毒标志物的LTR克隆支持免疫细胞、神经胶质细胞和骨髓来源的细胞类型中瞬时表达的能力，并建立LTR功能和先前目标中建立的临床参数之间的相关性。这些研究可能会导致发现更多工具来预测 HIVD 的发展，进而提供更多信息来指导 HIV-1 感染患者的治疗管理。