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

基本信息

批准号：
10414084
负责人：
Teresa Evering
金额：
$ 21.45万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2024-05-31
项目状态：
已结题

项目摘要

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.

项目摘要/摘要尽管临床护理取得了进步，但长期抗逆转录病毒疗法（ART）患有HIV-1（PLWH）的人仍在存在认知和行为缺陷以及HIV-1相关神经认知疾病的流行率（手）为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，并且难以使用当前方法研究。在此提案中，我们将使用生理上的相关，研究HIV-1中神经元衰老的可修改机械途径，参与者衍生的细胞模型。近年来，从已经证明了患者来源的成纤维细胞。与未成熟的神经元种群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。结果，这些INS代表了主要的技术进步。我们的初步数据证明了我们从PLWH的皮肤活检中产生INS的能力。据我们所知，这使我们第一个将该技术应用于HIV-1中神经元健康的小组。我们现在建议使用这些工具确定基因表达的差异是否反映了加速的年龄和/或其他HIV疾病 - 通过源自源自的INS的转录表型可以明显看出相关的转录特征年龄匹配的年龄较大和年龄较大的HIV阴性和HIV阳性队列。 INS的功能分析将测试神经元衰老的假设病理机理。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，已知越过血脑屏障学位可能对神经元健康具有类和/或药物特异性的有害影响。因此，我们在探索性二级分析，假设存在可能修改的机械途径 HIV-1的神经元损伤也依赖于ARV类。我们将以两个特定的方式研究我们的假设目标。在AIM 1中，我们将从临床表征良好的参与者中产生患者来源的INS。我们将根据Mertens等人的生成INS。（细胞干细胞，2015年），来自6个临床定义良好的队列的皮肤活检按年龄，HIV-1状态和使用蛋白酶抑制剂（PI）与整合酶链分层基于转移抑制剂（Insti）的艺术。在AIM 2中，我们将确定与神经元相关的基因表达通过年龄，HIV-1状态和手推车使用的途径通过从良好的转录进行转录分析定义的临床队列。差异基因表达分析将在跨人群中进行确定，年龄（旧与年轻），以及在次要探索性分析中，HIV-1（正面与负）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的核细胞质隔室化（NCC）的损害表征了来自较老的HIV-1阳性队列的INS。我们的小说将此INS技术应用于PLWH的翻译研究，使我们有机会填补我们的空白了解HIV-1中神经元衰老并确定改善认知的治疗靶标。