Leveraging Directly Reprogrammed Human Neurons to Investigate the Molecular Impact of Age and Distinct Antiretroviral Therapies in Individuals Living with HIV-1

利用直接重编程的人类神经元研究年龄和独特抗逆转录病毒疗法对 HIV-1 感染者的分子影响

• 批准号: 10257736
• 负责人: Teresa Evering
• 金额: $ 27.43万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10257736
• 关键词: Adverse effects; Age; Age-Years; Aging; Anti-Retroviral Agents; Autophagocytosis; Behavioral; Biological; Biological Assay; Biopsy; Blood - brain barrier anatomy; Carrier Proteins; Cell model; Characteristics; Clinical; Cognition; Cognitive; Collaborations; Complex; Confounding Factors (Epidemiology); Control Groups; Data; Defect; Dermal; Development; Disease; Down-Regulation; Elderly; Enrollment; Fibroblasts; Flow Cytometry; Functional disorder; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic Transcription; HIV; HIV Seronegativity; HIV Seropositivity; HIV-1; HIV-associated neurocognitive disorder; Health; Human; Impaired cognition; Impairment; In Vitro; Incidence; Individual; Inflammation; Injury; Integrase; Memorial Sloan-Kettering Cancer Center; Methods; Molecular; Morphology; Neuraxis; Neurocognitive; Neurocognitive Deficit; Neuroglia; Neuronal Injury; Neurons; Neuropsychological Tests; Nuclear Pore; Ontology; Oxidative Stress; Participant; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Physiological; Population; Prevalence; Protease Inhibitor; Rattus; Regimen; Reporter; Reporting; Sampling; Skin; Stem Cell Research; System; Technology; Testing; Tissue-Specific Gene Expression; Validation; age effect; age related; antiretroviral therapy; base; clinical care; cohort; comorbidity; human disease; immunocytochemistry; improved; induced pluripotent stem cell; inhibitor/antagonist; interest; mitochondrial dysfunction; neurotrophic factor; novel; research facility; secondary analysis; sex; stem cells; therapeutic target; tool; transcriptome sequencing; translational study

PROJECT SUMMARY/ABSTRACT Despite advances in clinical care, people living with HIV-1 (PLWH) on long-term antiretroviral therapy (ART) still present with cognitive and behavioral deficits and the prevalence of HIV-1 associated neurocognitive disorders (HANDs) is 30%-50%. The mechanisms underlying HIV-induced central nervous system (CNS) dysfunction in these patients are complex, multifactorial and difficult to study using current methods. In this proposal, we will investigate modifiable mechanistic pathways of neuronal aging in HIV-1 using physiologically relevant, participant-derived cell models. In recent years, the ability to generate directly induced neurons (iNs) from patient-derived fibroblasts has been demonstrated. Unlike the immature neuronal populations generated from induced pluripotent stem cells (iPSCs), iNs retain neuron-specific, aging-associated gene-expression characteristics of the donor. As a result, these iNs represent a major technological advance. Our preliminary data demonstrates our ability to generate iNs from the skin biopsies of PLWH. To our knowledge, this makes us the first group to apply this technology to study of neuronal health in HIV-1. We now propose to use these tools to determine if differences in gene expression reflective of accelerated age- and/or additional HIV disease- associated transcriptional signatures are evident through the transcriptional phenotyping of iNs derived from age-matched young and older HIV-negative and HIV-positive cohorts. Functional analyses of the iNs will test hypothesized pathomechanisms of neuronal aging. A growing body of scientific evidence also suggests that antiretroviral drugs (ARVs) widely used in HIV-1 therapy and known to cross the blood-brain barrier to varying degrees may have class- and/or drug-specific deleterious effects on neuronal health. We therefore, in exploratory secondary analyses, hypothesize that there are potentially modifiable, mechanistic pathways of neuronal injury in HIV-1 that are also ARV class dependent. We will investigate our hypotheses in two specific aims. In AIM 1, we will generate patient-derived iNs from clinically well-characterized participants. We will generate iNs as per Mertens et al. (Cell Stem Cell, 2015) from the skin biopsies of 6 clinically well-defined cohorts stratified by age, HIV-1 status and use of protease-inhibitor (PI)- versus Integrase strand transfer inhibitor (INSTI)-based ART. In AIM 2, we will determine neuronal-associated gene-expression pathways modulated by age, HIV-1 status and cART usage through transcriptional profiling of iNs from well- defined clinical cohorts. Differential gene expression analysis will be performed across cohorts in attempts to determine, age (old vs. young), and in secondary exploratory analyses, HIV-1 (positive vs. negative) and ARV (PI vs. INSTI) effects on gene transcription. We will then determine if age-dependent nucleocytoplasmic compartmentalization (NCC) impairments characterize iNs from older, HIV-1 positive cohorts. Our novel application of this iNs technology to the translational study of PLWH allows us the opportunity to fill a gap in our understanding of neuronal aging in HIV-1 and identify therapeutic targets for improved cognition.

项目概要/摘要 尽管临床护理取得了进步，但接受长期抗逆转录病毒治疗 (ART) 的 HIV-1 感染者 (PLWH) 仍然 存在认知和行为缺陷以及 HIV-1 相关神经认知障碍的流行 （手）是30%-50%。 HIV引起的中枢神经系统（CNS）功能障碍的机制 这些患者是复杂的、多因素的，并且难以使用现有方法进行研究。在本提案中，我们将 利用生理相关性研究 HIV-1 中神经元衰老的可修改机制途径， 参与者衍生的细胞模型。近年来，直接产生诱导神经元（iN）的能力 患者来源的成纤维细胞已被证实。与产生的未成熟神经元群体不同 诱导多能干细胞 (iPSC)、iNs 保留神经元特异性、与衰老相关的基因表达 捐赠者的特征。因此，这些 iN 代表了一项重大技术进步。我们的初步 数据证明了我们从感染者皮肤活检中生成 iN 的能力。据我们所知，这使我们 第一个应用该技术来研究 HIV-1 神经元健康的小组。我们现在建议使用这些工具 确定基因表达差异是否反映了年龄增长和/或其他艾滋病毒疾病 相关转录特征通过源自 iN 的转录表型显而易见 年龄匹配的年轻和老年艾滋病毒阴性和艾滋病毒阳性队列。 iN 的功能分析将测试 假设的神经元衰老的病理机制。越来越多的科学证据也表明 抗逆转录病毒药物 (ARV) 广泛用于 HIV-1 治疗，已知可穿过血脑屏障， 学位可能会对神经元健康产生类别和/或药物特异性的有害影响。因此，我们在 探索性二次分析，假设存在潜在可修改的机械途径 HIV-1 中的神经元损伤也与 ARV 类别相关。我们将在两个具体方面研究我们的假设 目标。在 AIM 1 中，我们将从临床特征良好的参与者中生成源自患者的 iN。我们将 根据 Mertens 等人生成 iN。 （Cell Stem Cell，2015）来自 6 个临床明确的队列的皮肤活检 按年龄、HIV-1 状态和蛋白酶抑制剂 (PI) 与整合酶链的使用进行分层 基于转移抑制剂（INSTI）的 ART。在 AIM 2 中，我们将确定神经元相关基因表达 通过对来自井的 iN 进行转录分析，发现年龄、HIV-1 状态和 cART 使用情况调节的途径 定义的临床队列。将跨队列进行差异基因表达分析，试图 确定年龄（年长与年轻），并在二次探索性分析中确定 HIV-1（阳性与阴性）和 ARV （PI 与 INSTI）对基因转录的影响。然后我们将确定年龄依赖性核细胞质是否 区室化（NCC）损伤是老年 HIV-1 阳性人群 iN 的特征。我们的小说 将这种 iNs 技术应用于 PLWH 的转化研究使我们有机会填补我们的空白 了解 HIV-1 中的神经元衰老并确定改善认知的治疗靶点。