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: 10563136
• 负责人: Joshua Charles Cyktor
• 金额: $ 72.32万
• 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/10563136
• 关键词: AIDS clinical trial group; Attention; Autopsy; Behavior; Binding; Biological; Biological Assay; Blood; CD4 Positive T Lymphocytes; Cell Lineage; Cells; Central Nervous System; 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; Length; Link; Living Donors; Measures; Mental Health; Methods; Microglia; Modeling; Molecular; Motor; Myeloid Cells; National Institute of Mental Health; Nature; Neuropsychology; Outcome; Participant; Peripheral Blood Mononuclear Cell; Persons; Phenotype; Plasma; Proviruses; Puncture procedure; RNA; Research Domain Criteria; Site; Sorting; 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; immunoregulation; 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 尽管艾滋病毒长期被抑制在抗逆转录病毒疗法(ART)中，但根除或持续缓解 尚未实现感染。在外周血单核细胞中仍可检测到低水平的艾滋病毒DNA 接受抗逆转录病毒治疗的艾滋病毒携带者(PWH)的细胞(PBMC)和含有反弹能力病毒的细胞可以 栖息在避难所组织部位，包括淋巴结、肠道、生殖道和中枢神经系统 (CNS)。在艾滋病临床试验小组内进行的一项研究中，我们最近使用了高度敏感的 活体献血者病毒学检测脑脊液细胞中HIV DNA含量高达50% 艺术。重要的是，那些可检测到脑脊液艾滋病毒DNA的人的整体认知功能比那些 脑脊液中未检出HIV DNA。我们随后发现，在CD4+T细胞中，HIV DNA浓度较高 来自脑脊液的细胞与同时代的PBMC以及包括髓系细胞在内的非典型细胞谱系的比较 可能在脑脊液中感染。最后，我们已经成功地使用单细胞转录切割法来鉴定独特的和 PWH患者脑脊液中罕见的细胞类型与HIV疾病状态相关，并进一步证明了 以确定富含PWH的细胞转录本与健康对照。对CNS HIV认识上的重大差距 持续性包括脑脊液中被感染的免疫细胞的特征和功能，是否为艾滋病毒 与血液中的前病毒相比，脑脊液中的前病毒是完整的，并且在基因上是区隔的，以及如何 这些特征与长期感染HIV的神经精神功能有关。由于脑脊液中存在的细胞数量是 低PWH抑制了ART，彻底的，同时描述了免疫学和病毒学 尚未实现中枢神经系统的景观。采用优化的腰椎穿刺术和新颖的 分子技术，我们克服了这些障碍，既严格地检测了细胞的表型 并对持续存在的HIV病毒的大小、稳定性、完整性和序列多样性进行了全面的表征 脑脊液与血液的比较。我们将使用单细胞技术来测量转录和细胞因子谱 结合新的完整HIV DNA定量和单基因组测序发现 CNS储集层内的新对比。最重要的是，我们建议严格审查认知 使用新的NIMH的复杂实施对艾滋病毒感染者的功能和心理健康的影响 研究领域标准(RDoC)框架，以及神经精神病学结果的差异如何与 不同人群中中枢神经系统的特异性免疫学和病毒学特征 神经精神共病的范围。