NEUROBIOLOGY

这是Masliah项目的修订申请，已被广泛修改以解决审查者的担忧：1)扩展我们的分析以包括体视学和定量实时聚合酶链式反应(qRT-PCR)，2)扩大我们对星形胶质细胞的神经病理学和体外研究，以及3)纳入新的初步数据，以支持更具创新性的概念，即由艾滋病毒和丙型肝炎病毒感染的细胞产生的干扰素(IFN)与甲氨蝶呤一起，可能增强可能促进神经变性的星形胶质细胞反应，并阻断神经营养因子，如成纤维细胞生长因子(FGFs)对Calbindin(CB)和 多巴胺能神经元。具体地说，我们假设新皮质和海马区CB-IR的丢失将与认知障碍有关，而黑质间质系统标志物的丢失将与运动和认知障碍相关。为了更新，并与PPG的其他组成部分合作，我们的目标是：1)阐明Meth+、HIV+病例脑内局部神经元损伤和星形胶质细胞的模式及其与HIVE的神经病理标志以及对皮质中间神经元和NS细胞选择性的干扰素和营养因子的水平的关系。2)探讨不同丙型肝炎病毒状态、不同脑脊液生物标志物和行为改变的吸毒HIV感染者脑内神经元损伤与星形胶质细胞增生症的关系。3)通过星形胶质细胞干扰素-α的变化，研究冰毒与HIV的协同神经毒性作用 IFI基因表达和神经元的成纤维细胞生长因子/SHH信号转导。分子机制的剖析将在神经细胞系、原代神经元培养和暴露于HIV蛋白的人胎脑聚集体中进行，无论存在或不存在Meth和FGF2，或FGFs和SHH。这些研究将有助于促进对艾滋病毒感染冰毒使用者认知和运动改变的细胞和分子机制的理解，并为这些复杂的疾病开发新的治疗策略。

This is a revised application for Project by Masliah that has been extensively modified to address the reviewers' concerns by: 1) expanding our analysis to include stereology and quantitative real time PCR (qRT-PCR), 2) broadening our neuropathological and in vitro studies of astroglial cells, and 3) incorporating new preliminary data in support of the more innovative concept that interferons (IFNs) produced by HIV and HCV-infected cells might, in conjunction with METH, enhance astroglial responses that might promote neurodegeneration and block the effects of neurotrophic factors such as fibroblast growth factors (FGFs) on calbindin (CB) and dopaminergic neurons. Specifically, we hypothesize that loss of CB-IR in the neocortex and hippocampus will be associated with cognitive deficits, while loss of markers in the nigrostiatal system will correlate with motor and cognitive deficits. For the renewal and in collaboration with other components of the PPG our aims are: 1) To elucidate patterns of regional neuronal damage and astrogliosis in the brains of METH+, HIV+ cases and their relationship with neuropathological markers of HIVE and levels of IFN expression and trophic factors selective for cortical interneurons and NS cells. 2} To determine the relationship between regional patterns of neuronal injury and astrogliosis in the brains of METH-using HIV infected cases who vary in HCV status, specific cerebrospinal fluid (CSF) biomarkers and behavioral alterations. 3) To investigate the synergistic neurotoxic effects of METH and HIV in vitro via alterations in astroglial IFN-alpha and IFI gene expression and neuronal FGF/SHH signaling. Dissection of the molecular mechanisms will be performed in neuronal cell lines, primary neuronal cultures, and human fetal brain aggregates exposed to HIV proteins in the presence or absence of METH and FGF2, or FGFS and SHH. These studies will help advance understanding of the cellular and molecular mechanisms of cognitive and motor alterations in HIV infected METH users and in developing novel treatment strategies for these complex disorders.