A Precision Medicine Approach to Investigating the Molecular Impact of Age and Neurocognitive Impairment in People Living with HIV (PLWH)

项目摘要 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 行为缺陷和HIV-1相关神经认知障碍（HANDs）的患病率为30%-50%。的 这些患者中HIV诱导的中枢神经系统（CNS）功能障碍的潜在机制是复杂的 用现有的方法进行研究具有挑战性。在这个建议中，我们将研究可修改的机制， 使用个性化的，生理相关的，参与者的PLWH神经认知障碍的途径， 衍生细胞模型。近年来，从患者源性神经元产生直接诱导神经元（iN）的能力受到了广泛关注。 成纤维细胞已被证实。与诱导产生的未成熟神经元群体不同， 多能干细胞（iPSC），iNs保留了神经元特异性，衰老相关基因表达特征， 捐赠者。因此，这些iN代表了一项重大的技术进步。我们的初步数据显示 我们从PLWH的皮肤活检中产生iN的能力。据我们所知，这使我们成为第一批 将这项技术应用于研究HIV-1的神经健康。我们现在建议使用这些工具来确定， 反映神经认知障碍的基因表达的差异通过转录调控是明显的。 来源于年龄、性别和种族匹配的PLWH队列的iN的表型分析（富集> 60岁的个体） 年龄。iN的功能分析将测试假设的神经元功能障碍的病理机制。我们将 在两个特定的目标中研究我们的假设。在AIM 1中，我们将从临床上生成患者来源的iN。 良好的参与者。我们将根据Mertens等人（Cell Stem Cell，2015）从皮肤中生成iN 根据神经认知程度分层的3个临床明确定义的低共病队列的活检 通过综合神经心理学检查和HIV-1确定的NCI。在AIM 2中，我们 将确定由神经认知障碍调节的神经元相关基因表达途径， 通过明确定义的临床队列的iN的转录谱分析的HIV-1状态。差异基因表达 将在队列中进行分析，以确定神经认知（正常vs.受损）， HIV-1（阳性与阴性）对基因转录的影响。体外实验将检验这一假设， 核质区室化损伤和衍生iNs中的凋亡与 供体NCI的程度。我们将这种iNs技术应用于PLWH的翻译研究， 我们有机会填补我们对HIV-1神经元功能障碍的理解的空白，并确定治疗方法。 提高认知能力的目标。

PROJECT ABSTRACT 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 and challenging to study using current methods. In this proposal, we will investigate modifiable mechanistic pathways of neurocognitive impairment in PLWH using personalized, 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 neurocognitive impairment are evident through the transcriptional phenotyping of iNs derived from age, sex and race-matched cohorts of PLWH enriched for individuals > 60 years of age. Functional analyses of the iNs will test hypothesized pathomechanisms of neuronal dysfunction. 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 3 clinically well-defined cohorts with low comorbid conditions stratified by degree of neurocognitive impairment (NCI) as determined by comprehensive neuropsychological examination and HIV-1. In AIM 2, we will determine neuronal-associated gene-expression pathways modulated by neurocognitive impairment and HIV-1 status through transcriptional profiling of iNs from well-defined clinical cohorts. Differential gene expression analysis will be performed across cohorts in attempts to determine, neurocognitive (normal vs. impaired), and HIV-1 (positive vs. negative) effects on gene transcription. In vitro experiments will test the hypothesis that impairments in nucleocytoplasmic compartmentalization and apoptosis in derived iNs positively correlate with the degree of donor NCI. 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 dysfunction in HIV-1 and identify therapeutic targets for improved cognition.