High Specificity HIV-1 Markers Predictive of Neuro-AIDS

描述(申请人提供)：人类免疫缺陷病毒1型(HIV-1)相关疾病在免疫和中枢神经系统的进展与病毒在这些间隔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，在病毒复制过程中引入的HIV-1基因组内的基因改变可能与HIV-1疾病的阶段和/或神经状态有关。我们最近已经证明，病毒反式激活蛋白VPR与特定的HIV-1 LTRC/EBP结合位点配置具有增强的亲和力，可以导致增强的长末端重复序列(LTR)激活，并与终末期疾病和HIVD相关。更多的研究已经确定了一些HIV-1 LTR序列特征，这些特征在进展性疾病期间外周血腔内的频率增加，并跟踪HIVD的发展。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，在HIV-1疾病过程中，在病毒复制过程中选择的LTR内的C/EBP和Sp结合位点签名，可以用作分子标记来识别可能更容易发生HIVD的HIV-1感染者。本申请的具体目的是：(1)建立HIV-1LTR克隆库和序列数据库，来源于HIV-1感染的外周血液中的免疫细胞群，纵向收集自有明确的抗逆转录病毒治疗、滥用药物和神经状况的患者，以及尸检时从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3T C/EBP位点I和5TSp位点III标记(以及其他LTR区域中的潜在标记)的存在，并建立标记患病率与疾病进展和严重程度、神经状态、抗逆转录病毒治疗和滥用药物使用之间的关联；(3)建立单核苷酸多态(SNP)基因分析，用于检测3T C/EBP位点I和5T Sp位点III标记(以及LTR其他区域中的潜在标记)，以用作指示和/或预测周围疾病进展和神经状态的诊断分析；以及(4)检测含有病毒标志物(S)的LTR克隆在免疫、神经胶质和骨髓来源的细胞类型中支持瞬时表达的能力，并建立LTR功能与先前目的建立的临床参数之间的相关性。这些研究可能导致确定其他工具来预测HIVD的发展，并反过来提供更多信息来指导HIV-1感染患者的治疗管理。

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.