Routes to enhanced HIV neuropathogenesis through expression of subclinical levels of endogenous amyloid-beta

这是艾滋病毒和阿尔茨海默病（AD）实验室之间的合作项目的重新提交， 研究HIV感染和β淀粉样蛋白（A β）（AD的标志）在促进记忆方面的潜在协同作用 损伤接受抗逆转录病毒治疗的艾滋病毒感染者中，有一半会出现神经认知障碍（NCI）， 艾滋病毒的大脑负担。年龄是影响NCI进展的最一致因素。我们复制了HIV-NCI 感染EcoHIV（一种小鼠嗜性HIV）的常规小鼠的发病机制。MPI项目阿兰西奥博士 报道了非突触毒性剂量的外源性寡聚A β（oA β）和Tau，另一种AD标志， 在小鼠的突触功能障碍和记忆障碍中协同作用，表明AD蛋白具有致病性 在亚临床浓度下具有潜力。我们修订后的初步结果显示， 多种形式的EcoHIV和oA β，包括在AD小鼠模型的EcoHIV感染后， 下降模型包括家族性AD（FAD）转基因APP/PS1小鼠模型和两种迟发性AD 在一些实施方案中，使用表达人源化AD蛋白的LOAD小鼠模型。我们认为HIV和亚临床oA β过程 协同破坏突触可塑性，使HIV-NCI恶化，超过单独使用每种药物。具体目标 1）在FAD和LOAD小鼠模型中，在其发病之前，表征增强的HIV-NCI发病机制， AD症状。(A)优化临床前APP/PS1小鼠中EcoHIV感染的记忆缺陷的发作;（B） 通过海马LTP、突触-树突状突起、突触-树突状突起和突触-树突状突起来确定感染APP/PS1小鼠的疾病发作时的HIV-NCI严重程度。 损伤、凋亡、APP加工和淀粉样蛋白病理学（C）通过EcoHIV感染进行类似的研究， 表达各种人源化AD基因的LOAD模型小鼠，并选择一种LOAD模型用于进一步研究2） 使用APP/PS1小鼠和一种LOAD毒株，在EcoHIV中进行下一代RNA测序（RNA-seq） 感染的小鼠在达到HIV疾病状态后（A）产生来自海马（HPC）、前额叶（PC）、 皮质（PFC）和纹状体（STR），并定义与记忆缺陷特别相关的生物过程 B）通过HPC和STR提取物中的RT-QPCR或通过 在脑切片中进行免疫染色;以及C）在HPC中进行有限的核-RNA-seq分析以分配改变的HIV-1基因。 oA β加工/转录到特定细胞类型。3)研究负责促进艾滋病毒的分子机制- 小鼠中自然产生的亚临床水平人oA β背景下的NCI发病机制。(A)由于两 HIV和A β干扰wnt信号传导并导致树突修剪，研究HIV感染骨髓细胞的工作原理 与A β在树突完整性中的Wnt功能失调有关。B）由于EcoHIV下调CaMK II和CREB 在小鼠中研究潜在的A β-HIV会聚在oA β调节异常的CREB中的突触强化作用。 研究将包括EcoHIV感染中度和迟发性AD小鼠，学习和记忆测试，HPC 长时程增强、HPC突触树突完整性、凋亡、APP加工和淀粉样蛋白病理学 系统和大脑HIV负担，小鼠大脑和选择细胞类型的生物信息学。

This is a resubmission of a collaborative project between HIV and Alzheimer’s Disease (AD) laboratories to investigate potential synergy between HIV infection and amyloid β (Aβ), a hallmark of AD, in promoting memory impairment. Half of HIV patients on antiretroviral therapy develop neurocognitive impairments (NCI) despite low HIV brain burdens. Age is the most consistent factor affecting NCI progression. We reproduced HIV-NCI pathogenesis in conventional mice infected with EcoHIV, a mouse-tropic HIV. The Project MPI, Dr. Arancio, reported that non-synaptotoxic doses of exogenous oligomeric Aβ (oAβ) and Tau, another AD hallmark, cooperate in synaptic dysfunction and memory impairment in mice, suggesting that AD proteins have pathogenic potential at their subclinical concentrations. Our revised preliminary results show such cooperation between EcoHIV and oAβ in multiple formats including after EcoHIV infection of AD mouse models prior to their cognitive decline. The models included transgenic APP/PS1 mouse model of familial AD (FAD) and two late onset AD (LOAD) mouse models expressing humanized AD proteins. We propose that HIV and subclinical oAβ processes cooperate in disrupting synaptic plasticity to exacerbate HIV-NCI beyond each agent alone. The Specific Aims are to: 1) Characterize enhanced HIV-NCI pathogenesis in FAD and LOAD mouse models prior to onset of their AD symptoms. (A) optimize the onset of memory deficits with EcoHIV infection in preclinical APP/PS1 mice; (B) determine HIV-NCI severity at disease onset by infected APP/PS1 mice by hippocampal LTP, synaptodendritic injury, apoptosis, APP processing, and amyloid pathology (C) Conduct similar studies by EcoHIV infection of LOAD model mice expressing various humanized AD genes and select one LOAD model for further studies 2) Using APP/PS1 mice and one LOAD strain, conduct next generation RNA sequencing (RNA-seq) in EcoHIV infected mice upon reaching HIV disease state (A) generate transcriptome from hippocampus (HPC), prefrontal cortex (PFC), and striatum (STR), and define biological processes specifically associated with memory defects caused jointly by HIV and oAβ; B) confirm some gene modulation by RT-QPCR in HPC and STR extracts or by immunostaining in brain section; and C) conduct limited nuclear-RNA-seq analysis in HPC to assign altered HIV- oAβ processes/transcripts to specific cell types. 3) Study molecular mechanisms responsible for promoting HIV- NCI pathogenesis in the setting of naturally produced subclinical levels of human oAβ in mice. (A) since both HIV and Aβ disturb wnt signaling and cause dendritic pruning, examine how HIV infection of myeloid cells works with Aβ in dysregulating Wnt function in dendritic integrity. B) since EcoHIV downmodulates CaMKII and CREB in mice study potential Aβ-HIV convergence in oAβ dysregulation of the CREB role in synaptic strengthening. Studies will include EcoHIV infection of moderate and late onset AD mice, tests of learning and memory, HPC long-term potentiation, HPC synaptodendritic integrity, apoptosis, APP processing, and amyloid pathology system and brain HIV burdens, bioinformatics of mouse brain and select cell types.