CNS Viral Persistence and Neuropsychiatric Perturbations in HIV: Single cell and Molecular Interrogation

HIV 中的中枢神经系统病毒持久性和神经精神扰动：单细胞和分子审讯

• 批准号: 10395614
• 负责人: Joshua Charles Cyktor
• 金额: $ 73.15万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10395614
• 关键词: AIDS clinical trial group; Attention; Autopsy; Behavior; Biological; Biological Assay; Blood; CD4 Positive T Lymphocytes; Cell Lineage; Cells; Central Nervous System Diseases; Cerebrospinal Fluid; Characteristics; Clinical; Clonality; Cognitive; Complex; Coupled; DNA; Data; Detection; Disease; Disease associated microglia; Disease remission; Emotions; Frequencies; Functional disorder; Gene Expression Profile; Genetic Transcription; Genetic Variation; Goals; HIV; HIV Infections; HIV Seronegativity; HIV-1; Immune; Immunologic Factors; Immunologics; Infection; Inflammatory; Lead; Length; Link; Living Donors; Measures; Mental Health; Methods; Microglia; Modeling; Molecular; Motor; Myeloid Cells; National Institute of Mental Health; Nature; Neuraxis; Neuropsychology; Outcome; Participant; Peripheral Blood Mononuclear Cell; Persons; Phenotype; Plasma; Proviruses; Puncture procedure; RNA; Research Domain Criteria; Site; Spinal Puncture; System; Techniques; Time; Tissues; Transcript; Viral; Virus; antiretroviral therapy; brain tissue; cell type; clinically significant; cognitive ability; cognitive control; cognitive function; cognitive neuroscience; cohort; combinatorial; comorbidity; cytokine; executive function; genome sequencing; improved; inflammatory milieu; innovation; insight; integration site; lymph nodes; neuropsychiatry; novel; reproductive tract; single cell analysis; single cell technology; targeted treatment; transcriptomics

Project Abstract Despite prolonged suppression of HIV on antiretroviral therapy (ART), eradication or sustained remission of the infection has not been achieved. Low levels of HIV DNA are still detectable in peripheral blood mononuclear cells (PBMC) from people living with HIV (PWH) taking ART, and cells containing rebound-competent virus can reside in sanctuary tissue sites, including lymph nodes, gut, genital tract and the central nervous system (CNS). In a study conducted within the AIDS Clinical Trials Group, we have recently used highly sensitive virologic assays in living donors to detect HIV DNA in cerebrospinal fluid (CSF) cells in up to 50% on long-term ART. Importantly, those with detectable CSF HIV DNA had poorer global cognitive function that those in whom CSF HIV DNA was not detected. We have subsequently shown higher concentrations of HIV DNA in CD4+ T cells from CSF compared to contemporaneous PBMC, and that atypical cell lineages including myeloid cells may be infected in CSF. Finally, we have successfully used single cell transcriptomics to identify unique and rare cell types in the CSF in PWH associated with HIV disease status and have further demonstrated the ability to identify cellular transcripts enriched in PWH versus healthy controls. Critical gaps in understanding CNS HIV persistence include what the characteristics and function of infected immune cells are in CSF, whether HIV proviruses in CSF are intact and genetically compartmentalized compared to proviruses in blood, and how these features relate to neuropsychiatric function of long-term HIV. Since the number of cells present in CSF is low in PWH suppressed on ART, thorough, simultaneous characterization of the immunologic and virologic landscape of the CNS has not been achieved. Using optimized lumbar puncture procedures and novel molecular techniques, we have overcome these obstacles to both rigorously examine the phenotype of CSF cells and thoroughly characterize the size, stability, intactness, and sequence diversity of persistent HIV in CSF compared to blood. We will use single cell technology to measure the transcriptional and cytokine profile of CNS cells combined with novel quantification of intact HIV DNA and single genome sequencing to discover new correlations within the CNS reservoir. Most importantly, we propose to rigorously examine the cognitive function and mental health of people living with HIV using sophisticated implementation of the new NIMH Research Domain Criteria (RDoC) framework, and how differences in neuropsychiatric outcomes relate to specific immunological and virological characteristics of the CNS in a diverse cohort of participants with a range of neuropsychiatric comorbidity.

项目摘要 尽管长期抑制了抗逆转录病毒疗法（ART）的HIV，但消除或持续缓解 尚未实现感染。低水平的HIV DNA在外周血单核中仍可检测到 来自艾滋病毒（PWH）服用ART的患者的细胞（PBMC）和含有反弹性病毒的细胞可以 居住在庇护组织部位，包括淋巴结，肠道，生殖道和中枢神经系统 （CNS）。在AIDS临床试验组中进行的一项研究中，我们最近使用了高度敏感的 在活体供体中的病毒学测定法检测脑脊液（CSF）细胞中高达50％的HIV DNA。 艺术。重要的是，患有CSF HIV DNA的人的全球认知功能较差 未检测到CSF HIV DNA。随后，我们在CD4+ T中显示了较高浓度的HIV DNA 与同时PBMC相比，CSF的细胞以及包括髓样细胞在内的非典型细胞谱系 可以在CSF中感染。最后，我们成功使用了单细胞转录组学来识别独特的和 与HIV疾病状况相关的PWH中，CSF中的罕见细胞类型，并进一步证明了能力 确定富含PWH与健康对照的细胞转录本。了解中枢神经系统艾滋病毒的关键差距 持久性包括感染免疫细胞在CSF中的特征和功能，无论是HIV是否 与血液中的病毒相比 这些特征与长期HIV的神经精神功能有关。由于CSF中存在的细胞数为 抑制ART，彻底的，同时表征免疫学和病毒学的PWH中的PWH低。 中枢神经系统的景观尚未实现。使用优化的腰椎穿刺程序和新颖 分子技术，我们克服了这些障碍，以严格检查 CSF细胞并彻底表征了持续HIV的大小，稳定性，完整性和序列多样性 与血液相比，CSF。我们将使用单细胞技术来测量转录和细胞因子谱 中枢神经系统细胞与完整的HIV DNA和单个基因组测序的新型定量结合在一起发现 中枢神经系统储层内的新相关性。最重要的是，我们建议严格检查认知 使用新NIMH的复杂实施艾滋病毒的人的功能和心理健康 研究领域标准（RDOC）框架，以及神经精神诊断的差异与 中枢神经系统的特定免疫学和病毒学特征在不同的参与者中 神经精神病合并症的范围。