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: 10258495
• 负责人: Joshua Charles Cyktor
• 金额: $ 72.18万
• 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/10258495
• 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 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; 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; virology

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 尽管通过抗逆转录病毒疗法（ART）长期抑制艾滋病毒，但艾滋病毒的根除或持续缓解 感染尚未实现。外周血单核细胞中仍可检测到低水平的 HIV DNA 来自接受 ART 的 HIV 感染者 (PWH) 的细胞 (PBMC)，以及含有具有反弹能力的病毒的细胞可以 存在于庇护组织部位，包括淋巴结、肠道、生殖道和中枢神经系统 （中枢神经系统）。在艾滋病临床试验组进行的一项研究中，我们最近使用了高度敏感的 对活体捐献者进行病毒学检测，长期检测脑脊液 (CSF) 细胞中高达 50% 的 HIV DNA 艺术。重要的是，那些脑脊液中可检测到 HIV DNA 的人的整体认知功能比那些携带可检测到的脑脊液 HIV DNA 的人要差。 脑脊液中未检测到 HIV DNA。随后我们发现 CD4+ T 中 HIV DNA 浓度更高 来自 CSF 的细胞与同期 PBMC 相比，以及包括骨髓细胞在内的非典型细胞谱系 可能被脑脊液感染。最后，我们成功地使用单细胞转录组学来识别独特且 感染者脑脊液中的稀有细胞类型与 HIV 疾病状态相关，并进一步证明了这种能力 鉴定富含 PWH 与健康对照的细胞转录本。理解中枢神经系统艾滋病毒的关键差距 持久性包括脑脊液中受感染免疫细胞的特征和功能，是否有 HIV 与血液中的原病毒相比，脑脊液中的原病毒是完整的，并且在基因上是分开的，以及如何 这些特征与长期艾滋病毒的神经精神功能有关。由于脑脊液中存在的细胞数量为 ART 抑制的 PWH 低，全面、同时表征免疫学和病毒学 CNS 的景观尚未实现。采用优化的腰椎穿刺程序和新颖的 分子技术，我们克服了这些障碍，严格检查了表型 CSF 细胞并彻底表征持续性 HIV 的大小、稳定性、完整性和序列多样性 脑脊液与血液相比。我们将使用单细胞技术来测量转录和细胞因子谱 CNS 细胞结合完整 HIV DNA 的新颖定量和单基因组测序来发现 中枢神经系统储库内的新相关性。最重要的是，我们建议严格检查认知 使用新 NIMH 的复杂实施来改善艾滋病毒感染者的功能和心理健康 研究领域标准 (RDoC) 框架，以及神经精神结果的差异如何与 在不同的参与者群体中，中枢神经系统的特定免疫学和病毒学特征 神经精神合并症的范围。