Motor Dysfunction in cART-era HIV: Neural Circuitry and Pathogenesis

摘要 运动功能障碍在联合抗逆转录病毒治疗(CART)时代的HIV+人群中很普遍，然而，其 病因尚不清楚，因为脑血管疾病可能是其病因之一。我们建议研究 HIV相关马达的光谱、潜在神经回路和细胞类型特定的分子特征 功能障碍，有以下目的和假设：目的1.识别与运动相关的神经区域 伴有和不伴有脑血管疾病的HIV+患者的损害。160名接受CART治疗的受试者将 被招募到结构和功能磁共振成像研究，以测试假设 运动任务表现的神经解剖学基础将因运动状态(正常/异常)和存在或 无脑血管合并症。在这项2x2分析中，曼哈顿艾滋病毒脑库的参与者 (MHBB)将形成招募的核心，对运动功能进行多模式评估。这些分析将 在第一年和第二年进行。然后，在确定了与运动功能障碍关系最密切的区域后， 在第三年到第五年，我们将检查MHBB队列中的尸检大脑：目标2。细胞类型特定 背外侧纹状体、腹侧中脑和选定的灰色和白色的转录组和表观基因组图谱 目标1中定义的感兴趣物质区域(ROI)，以识别运动功能障碍的分子特征。我们的 假设运动功能障碍的分子起源可以通过区域和细胞类型来阐明。 转录组和开放染色质相关的组蛋白乙酰化和甲基化的特异性分析 风景。具体地说，HIV+脑区神经元、星形胶质细胞和少突胶质细胞特征 运动功能受累会受到HIV炎症负担和局部脑血管疾病的影响； 而运动功能将通过这些区域变化来预测。荧光激活的原子核分选 涉及运动功能障碍的神经解剖区域将用于细胞类型的特定输入部分 基因组规模的RNA-Seq和组蛋白芯片-Seq的材料。在连续的组织中，艾滋病毒DNA的检测通过 巢式聚合酶链式反应，单核细胞/小胶质细胞CD68和CD163免疫组织化学激活，以及脑 对小血管病变(CSVD)进行小动脉壁增厚的形态计量学分析。100 MHBB HIV+大脑将根据认知特征和血管风险进行选择，以获得最接近的生存 目标1中的受试者，以及通过随后的器官捐赠获得的受试者的大脑。50 人口统计学上相似的艾滋病毒大脑也将被研究。马达分子签名中的候选基因 有无血管疾病的功能障碍将通过qPCR进行测试。带着这些目标，我们将阐明 CART治疗的HIV患者运动功能障碍的神经回路，脑血管对其发生的贡献，以及 描述它的区域神经基因组学。对分子变化的洞察和改进的目标将是 与艾滋病毒人群中具有血管作用的更广泛的神经退行性疾病有关。

Summary Motor dysfunction is prevalent in combination antiretroviral therapy (cART) era HIV+ populations, however, its genesis is unclear, as cerebrovascular disease is likely to contribute to its etiology. We propose to study the spectrum, underlying neural circuitry, and cell type-specific molecular signatures of HIV-associated motor dysfunction, with the following aims and hypotheses: Aim 1. Identify neural regions associated with motor impairment in HIV+ patients with and without cerebrovascular disease. 160 cART-treated subjects will be recruited to a structural and functional magnetic resonance imaging study, to test the hypothesis that the neuroanatomical basis of motor task performance will vary by motor status (normal/abnormal) and presence or absence of cerebrovascular comorbidity. For this 2x2 analysis, participants in the Manhattan HIV Brain Bank (MHBB) will form a nidus for recruitment, with multimodal assessment of motor function. These analyses will be conducted in years 1 and 2. Then, having identified regions most strongly implicated in motor dysfunction, in years 3 through 5 we will examine autopsy brains from the MHBB cohort for: Aim 2. Cell-type specific transcriptome and epigenome mapping in dorsolateral striatum, ventral midbrain, and selected gray and white matter regions of interest (ROI) as defined in aim 1 to identify molecular signatures of motor dysfunction. Our hypothesis is that the molecular genesis of motor dysfunction can be elucidated through regional and cell-type specific analysis of transcriptome and open chromatin-associated histone acetylation and methylation landscape. Specifically, that neuronal, astrocyte, and oligodendrocyte signatures in HIV+ brain regions implicated in motor function will be affected by HIV-inflammatory burden and regional cerebrovascular disease; and that motor function will be predicted by these regional changes. Fluorescence-activated nuclei sorting in neuroanatomic regions implicated in motor dysfunction will be used for cell-type specific fractions of input material for genome-scale RNA-Seq and histone ChIP-seq. In contiguous tissue, assays of HIV DNA by nested PCR, monocyte/microglial cell activation by CD68 and CD163 immunohistochemistry, and cerebral small vessel disease (CSVD) by morphometric analysis of arteriolar wall thickening, will be done. 100 MHBB HIV+ brains will be selected on the basis of cognitive profiles and vascular risk to best approximate living subjects in aim #1, along with brains accrued from imaged individuals through subsequent organ donation. 50 demographically similar HIV- brains will also be studied. Candidate genes in molecular signatures of motor dysfunction with and without vascular disease will be tested via qPCR. With these aims, we will elucidate the neural circuitry of motor dysfunction in cART-treated HIV, cerebrovascular contributions to its genesis, and characterize its regional neurogenomics. Insight into molecular alterations and targets for amelioration will be relevant to a wider spectrum of neurodegenerative disorders with vascular contributions in HIV- populations.