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（Lewy Body dementia，LBD）是一种谱系疾病，包括路易体痴呆（Dementia with Lewy bodies，DLB）和 帕金森病痴呆症（PDD）。这两种痴呆症的神经病理学特征都是阿尔法- 除了与阿尔茨海默氏病相关的可变病理外， 疾病-淀粉样蛋白-β（Ab）斑块和/或高磷酸化tau的神经纤维缠结（NFT）。之一 PDD和DLB之间最明显的区别是运动障碍的时间发生， 认知障碍这通常会对准确的诊断和合适的患者提出挑战。 临床试验的入组、患者护理和现有对症治疗。为了更好地理解 这两种痴呆时间进展我们建议利用患者来源的诱导多能干细胞 （iPSC）培养模型-提供个性化患者分析和药物筛选的疾病模型系统。 为了确保该模型系统准确地反映个体患者的疾病发病机制，我们提出 从我们进行尸检的个体中产生分化为神经元的iPSC 组织可用。这为我们提供了直接比较转录组学和疾病的独特机会 来自同一个体的脑组织和分化的iPSC-神经元的病理学结果。此外，我们能够 以时间方式监测和表征PDD与DLB相比的疾病进展。我们 假设来自PDD患者的iPSC神经元将在其颞叶疾病中显示表型差异 与DLB患者iPSC-神经元相比的进展。我们进一步假设，有相似的基因， 分化的iPSC-神经元和原代尸检组织之间的表达谱和疾病病理学 都是从同一个人身上得到的。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）。在目标2中，我们 使PDD和DLB iPSC（来自与Aim 1相同的个体）分化成皮质前脑神经元以产生 疾病特异性神经元转录组谱，并检查PD和痴呆相关疾病表型。 这些研究将首次研究PDD和DLB的转录组谱，检查细胞疾病 表型的时间的方式和解决的疾病机制LBD在这一频谱障碍。 此外，这项工作将为药物靶点鉴定提供新的机会， 对这两种疾病中的每一种都具有特异性的新的治疗剂和生物标志物。这将有助于更好地- 需要改进PDD和DLB患者的疾病诊断和管理。

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.