High Specificity HIV-1 Markers Predictive of Neuro-AIDS

描述（由申请方提供）：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中特定细胞群中定位和复制的能力相关。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。我们最近已经证明，病毒反式激活蛋白Vpr对特定的HIV-1 LTR C/EBP结合位点构型表现出增强的亲和力，这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。另外的研究已经导致鉴定了许多HIV-1 LTR序列特征，其在进行性疾病期间在外周血隔室内频率增加，并随着HIVD的发展而跟踪。这些意见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期间选择）可用作分子标记物，以识别可能更容易发生HIVD的HIV-1感染个体。本申请的具体目的是（1）建立HIV-1 LTR克隆库和序列数据库，所述克隆库和序列数据库来源于外周血中的HIV-1感染的免疫细胞群，所述外周血纵向收集自具有关于抗逆转录病毒治疗、药物滥用和神经状态的明确临床史的患者，以及来源于CNS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3 TC/EBP位点I和5 Tsp位点III标记的存在（和其他LTR区域的潜在标志物），并建立标志物患病率与疾病进展和严重程度、神经系统状态、抗逆转录病毒治疗和滥用药物之间的相关性;（3）建立3 T C/EBP位点I和5 T Sp位点III标记的单核苷酸多态性（SNP）基因型检测方法（和LTR的其他区域中的潜在标志物），用作诊断测定指示和/或或预测外周疾病进展和神经系统状态;和（4）测定含有病毒标志物的LTR克隆支持在免疫、神经胶质和骨髓来源的细胞类型中瞬时表达的能力，并建立LTR功能与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识别额外的工具来预测HIVD的发展，反过来，提供更多的信息来指导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.