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携带者(PLWH)仍存在认知和 行为缺陷和HIV-1相关神经认知障碍(手)的患病率为30%-50%。这个 这些患者中HIV引起的中枢神经系统(CNS)功能障碍的机制很复杂 使用现有方法进行研究具有挑战性。在这个提案中，我们将研究可修改的机械论 采用个性化、生理学相关的参与者--PLWH中神经认知障碍的途径 派生的细胞模型。近年来，由患者来源的直接诱导神经元(INS)的能力 成纤维细胞已经被证实。不同于由诱导产生的未成熟神经元群体 多能干细胞(IPSCs)，INS保留了神经元特异性的，衰老相关的基因表达特征 捐赠者。因此，这些INS代表着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INS的能力。据我们所知，这使我们成为第一个 将这项技术应用于HIV-1神经元健康的研究。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 从年龄、性别和种族匹配的PLWH队列中获得的INS的表型为60岁的个体 年纪大了。对INS的功能分析将检验神经元功能障碍的假想病理机制。我们会 从两个特定的目标来考察我们的假设。在目标1中，我们将从临床上生成患者派生的INS 很有特点的参与者。我们将按照Merten等人的要求生成INS。(细胞干细胞，2015)来自皮肤 根据神经认知程度分层的3个低共病的临床明确队列的活组织检查 由综合神经心理检查和HIV-1确定的损害(NCI)。在AIM 2中，我们 将确定神经认知障碍调节的神经元相关基因表达途径 通过对明确的临床队列中的INS进行转录分析，了解HIV-1的状况。差异基因表达 将在队列中进行分析，试图确定神经认知(正常与受损)和 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 衍生性INS核质区域化损伤与细胞凋亡呈正相关 供体NCI程度。我们将这种INS技术应用到PLWH的翻译研究中，使得 美国有机会填补我们对HIV-1神经元功能障碍的理解空白并确定治疗方法 改善认知的目标。

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.