Transcriptomic assessment of pathology in PD with dementia and dementia with Lewy Bodies using iPSC neurons and brain tissue of the same individuals

项目摘要 路易体痴呆(LBD)是一种谱系疾病，包括路易体痴呆(DLB)和 帕金森氏病痴呆(PDD)。这两种痴呆都有阿尔法-阿尔茨海默病的神经病理特征。 除了与阿尔茨海默氏症相关的各种病理变化外，所谓的路易小体中的突触核蛋白(a-syn)包涵体 疾病-淀粉样β蛋白(Ab)斑块和/或高磷化tau的神经原纤维缠结(NFT)。其中一个 PDD和DLB之间最明显的区别是相对于以下因素发生的运动障碍的时间 认知障碍。这往往会对准确的诊断以及相应的患者提出挑战 参加临床试验、病人护理和现有的对症治疗。为了更好地理解 这两种痴呆的时间进展我们建议利用患者来源的诱导多能干细胞 (IPSC)文化模型--提供个性化患者分析和药物筛选的疾病模型系统。 为了确保这个模型系统准确地反映单个患者的疾病发病机制，我们建议 为了从个体身上产生分化为神经元的IPSCs，我们也对这些个体进行了尸检 提供纸巾。这为我们提供了直接比较转录和疾病的独特机会 来自脑组织的病理和来自同一个体的分化的IPSC神经元。此外，我们还能够 以时间的方式监测和描述PDD与DLB相比的疾病进展。我们 假设PDD患者的IPSC神经元在其颞叶疾病中表现出表型差异 与DLB患者的IPSC神经元相比，进展。我们进一步假设有相似的基因 分化的IPSC神经元与初级尸检组织的表达谱和疾病病理 从相同的人那里获得的。为了验证这一假设，我们将进行单核多组学测序 (SnRNA-和ATAC序列)来自PDD、DLB和健康对照尸检脑组织(目标1)。在目标2中，我们将 分化PDD和DLBIPSCs(来自与目标1相同的个体)为皮质前脑神经元以产生 疾病特异性神经元转录组图谱，并检测帕金森病和痴呆相关疾病的表型。 这些研究将首次研究PDD和DLB的转录组特征，检查细胞疾病 以时间的方式表现型，并解决LBD在这种谱系障碍中的疾病机制。 此外，这项工作将为药物靶标识别提供新的机会，希望能够识别 针对这两种疾病的新疗法和生物标记物。作为回报，这将促进更多- PDD和DLB患者的疾病诊断和管理有待改进。

PROJECT ABSTRACT Lewy Body dementia (LBD) is a spectrum disease that includes dementia with Lewy bodies (DLB) and Parkinson’s disease dementia (PDD). The two dementias share neuropathological characteristics of alpha- synuclein (a-syn) inclusion in so called Lewy bodies, in addition to variable pathologies related to Alzheimer’s disease – amyloid-beta (Ab) plaques and/or neurofibrillary tangles (NFT) of hyperposphorylated tau. One of the most distinct differences between PDD and DLB is the temporal occurrence of motor impairments relative to cognitive impairments. This often challenges an accurate diagnosis and consequently appropriate patient enrollment in clinical trials, patient care and existing symptomatic treatment. To better understand the distinct temporal progression of these two dementias we propose to utilize patient-derived induced pluripotent stem cell (iPSC) culture models – a disease model system that offers personalized patient analyses and drug screening. To ensure that this model system accurately reflects the individual patient’s disease pathogenesis, we propose to generate iPSCs differentiated into neurons from individuals from which we also have postmortem autopsy tissue available. This provides us with the unique opportunity to directly compare transcriptomics and disease pathology from brain tissue and differentiated iPSC-neurons from the same individual. In addition, we are able to monitor and characterize disease progression in PDD compared to DLB in a temporal manner. We hypothesize that iPSC-neurons from PDD patients will show phenotypic differences in their temporal disease progression compared to DLB patient iPSC-neurons. We further hypothesize that there are similarities of gene expression profiles and disease pathologies between differentiated iPSC-neurons and primary autopsy tissue obtained from the same individuals. To test this hypothesis we will perform single nuclei multi-omics sequencing (snRNA- and ATAC seq) from PDD, DLB and healthy control autopsy brain tissues (Aim 1). In Aim 2, we will differentiate PDD and DLB iPSCs (from the same individuals as Aim 1) into cortical forebrain neurons to generate a disease-specific neuronal transcriptome profile and to examine PD and dementia-related disease phenotypes. These studies will for the first time study the transcriptome profile of PDD and DLB, examine cellular disease phenotypes in a temporal manner and address the disease mechanisms of LBD in this spectrum disorder. Additionally, this work will provide novel opportunities for drug target identification with the hope of identifying novel therapeutics and biomarkers specific for each of the two disorders. This in return will facilitate the much- needed improvement of disease diagnosis and management of PDD and DLB patients.