Table1_Single-Cell Transcriptomics Uncovers Cellular Heterogeneity, Mechanisms, and Therapeutic Targets for Parkinson’s Disease.XLSX

Objective: This study aimed to exploit cellular heterogeneity for revealing mechanisms and identifying therapeutic targets for Parkinson’s disease (PD) via single-cell transcriptomics.
Methods: Single-cell RNA sequencing (scRNA-seq) data on midbrain specimens from PD and healthy individuals were obtained from the GSE157783 dataset. After quality control and preprocessing, the principal component analysis (PCA) was presented. Cells were clustered with the Seurat package. Cell clusters were labeled by matching marker genes and annotations of the brain in the CellMarker database. The ligand–receptor networks were established, and the core cell cluster was selected. Biological functions of differentially expressed genes in core cell clusters were analyzed. Upregulated marker genes were identified between PD and healthy individuals, which were measured in 18 PD patients’ and 18 healthy individuals’ blood specimens through RT-qPCR and Western blotting.
Results: The first nine PCs were determined, which can better represent the overall change. Five cell clusters were identified, including oligodendrocytes, astrocytes, neurons, microglial cells, and endothelial cells. Among them, endothelial cells were the core cell cluster in the ligand–receptor network. Marker genes of endothelial cells possessed various biological functions. Among them, five marker genes (ANGPT2, APOD, HSP90AA1, HSPA1A, and PDE1C) were upregulated in PD patients’ than in healthy individuals’ endothelial cells, which were confirmed in PD patients’ than in healthy individuals’ blood specimens.
Conclusion: Our findings revealed that the cellular heterogeneity of PD and endothelial cells could play a major role in cell-to-cell communications. Five upregulated marker genes of endothelial cells could be underlying therapeutic targets of PD, which deserve more in-depth clinical research.

研究目的：本研究拟借助单细胞转录组学技术，利用细胞异质性解析帕金森病（Parkinson’s disease, PD）的发病机制并筛选潜在治疗靶点。
研究方法：从GSE157783数据集获取帕金森病患者与健康个体中脑组织标本的单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）数据。经质量控制与预处理后，开展主成分分析（principal component analysis, PCA）。采用Seurat软件包对细胞进行聚类，通过匹配CellMarker数据库中脑组织的注释信息与标记基因，对各细胞簇进行标注。构建配体-受体互作网络，并筛选得到核心细胞簇。分析核心细胞簇内差异表达基因的生物学功能。此外，招募18名帕金森病患者与18名健康对照个体，采集外周血标本，通过实时定量聚合酶链反应（RT-qPCR）与蛋白质印迹法（Western blotting）验证帕金森病患者与健康个体间的上调差异标记基因。
研究结果：本研究确定前9个主成分可最优反映整体转录组变化特征。共鉴定出5个细胞簇，分别为少突胶质细胞、星形胶质细胞、神经元、小胶质细胞与内皮细胞。其中，内皮细胞为配体-受体互作网络中的核心细胞簇。内皮细胞的标记基因参与多种生物学功能；进一步分析发现，ANGPT2、APOD、HSP90AA1、HSPA1A及PDE1C这5个内皮细胞标记基因在帕金森病患者的内皮细胞中呈上调表达，且该结果在帕金森病患者与健康对照的外周血标本中得到验证。
研究结论：本研究结果揭示了帕金森病的细胞异质性特征，且内皮细胞可能在细胞间通讯过程中发挥核心调控作用。本次鉴定得到的5个内皮细胞上调标记基因有望成为帕金森病潜在治疗靶点，值得开展更深入的临床研究。