Brain Structural and Functional Connectome in HIV-Associated Neuroinflammation

母基金RO 1 MH 118020利用新的方法来评估 与健康人相比，HIV感染者（PWH）的大脑结构和功能连接的轨迹 在炎症和凝血病的媒介物（可溶性CD 14和CD 163，D- 二聚体、可溶性组织因子和TF+单核细胞）。此外，父母补助金还旨在评估调解 结构和功能连接以及大脑皮层厚度对特定认知领域的影响。 来自父母资助的初步数据显示，PWH中的非经典单核细胞（NCMs）与 认知障碍脑小血管疾病（CSVD）患者的NCM水平最高 与没有CSVD相比。此外，单核细胞TF表达增加PWH相比，HC。 在这份行政补充文件中，我们建议将我们目前的调查扩展到研究老龄化的影响 和HIV对非经典单核细胞亚群和CSVD背景下TF表达的影响， 连通性和认知。我们的方法是利用基线临床、成像和存储的 入组的220例研究受试者的标本，以解决下列特定目的： 目的1：评估衰老和HIV对非经典单核细胞亚型的影响。我们假设 衰老和HIV将对NCM的细胞状态产生叠加或协同效应， 释放炎症因子，促进单核细胞-内皮细胞相互作用，这是单核细胞 转移到脑实质。将通过细胞索引评估NCM亚型， 转录组和表位测序（CITE-seq）。实验将在威尔斯亲王医院进行， 年龄≤40岁和≥50岁且无CSVD的健康对照研究受试者。 目的2：研究非经典单核细胞状态对CSVD的影响。我们假设 患有CSVD的PWH将具有独特的单核细胞亚型，其将使用CITE-seq进行评估， 使用我们小组和其他人开发的计算工具来表征。具体而言，CITE-seq将 在年龄≥50岁的PWH和健康对照研究参与者中进行，伴和不伴CSVD。 目的3：评估非经典单核细胞的细胞状态是否介导脑内的改变 连接性以及这是否反映在认知表现中。我们假设NCMs亚型 促进神经炎症，导致CSVD和改变大脑连接。临床相关的 CSVD和改变的大脑连接是认知障碍。我们将使用先前描述的 PathCellNet方法沿着加权基因相关网络分析（WGCNA），针对单基因组优化。 细胞分析，以确定可与临床相关的NCM亚型特异性基因特征 测量.我们还将使用结构方程模型（SEM）来理解NCM的因果效应 关于认知障碍

The parent grant RO1 MH118020 utilizes novel methodologies to assess the longitudinal changes in the trajectory of the brain structural and functional connectivity in people with HIV (PWH) compared to healthy controls (HC) in the context of intermediaries of inflammation and coagulopathy (soluble CD14 and CD163, D- dimer, soluble tissue factor and TF+ monocytes). Further, the parent grant also aims to assess the mediation effect of structural and functional connectivity and cerebral cortical thickness on specific cognitive domains. Preliminary data from the parent grant show that non-classical monocytes (NCMs) in PWH correlate with cognitive impairment. NCMs reach the highest levels in those with cerebral small vessel disease (CSVD) compared to no CSVD. Furthermore, monocyte TF expression is increased in PWH compared to HC. In this administrative supplement, we propose to extend our current investigations to study the effect of aging and HIV on non-classical monocyte subpopulations and TF expression in the context of CSVD, brain connectivity, and cognition. Our approach is to take advantage of the baseline clinical, imaging, and stored specimens of the 220 study participants enrolled to address the Specific Aims listed below: Aim 1: To assess the effect of aging and HIV on non-classical monocyte subtypes. We hypothesize that aging and HIV will have additive or synergistic effects on NCMs' cellular states characterized by pathways that release inflammatory factors and promote monocyte-endothelial cell interaction, an important step in monocyte transmigration to the brain parenchyma. NCMs subtypes will be assessed via Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-seq). The experiments will be conducted in PWH and healthy control study participants aged ≤40 and ≥50 without CSVD. Aim 2: To characterize the effect of cellular states of non-classical monocytes in CSVD. We hypothesize that PWH with CSVD will have unique subtypes of monocytes which will be assessed using CITE-seq and will be characterized using computational tools developed by our group and others. Specifically, CITE-seq will be performed on PWH and healthy control study participants aged ≥50, with and without CSVD. Aim 3: To assess whether cellular states of non-classical monocytes mediate alterations in brain connectivity and whether this is reflected in cognitive performance. We hypothesize that NCMs subtypes promote neuroinflammation, contributing to CSVD and altered brain connectivity. The clinical correlate of CSVD and altered brain connectivity is cognitive impairment. We will use our previously described PathCellNet approach along with Weighted Gene Correlation Network Analysis (WGCNA) optimized for single- cell analysis to define NCMs subtype-specific gene signatures that can be correlated with the clinical measurements. We will also use structural equation modeling (SEM) to understand the causal effect of NCMs on cognitive impairment.