中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (9): 2037-2046.doi: 10.4103/1673-5374.366493

• 原著:退行性病与再生 • 上一篇    下一篇

与核受体相关因子1缺陷导致帕金森病多巴胺能神经元损伤的分子机制：单核RNA测序分析

  

  • 出版日期:2023-09-15 发布日期:2023-03-07
  • 基金资助:
    国家自然科学基金项目（81971006）

Single-nuclei RNA sequencing uncovers heterogenous transcriptional signatures in Parkinson’s disease associated with nuclear receptor-related factor 1 defect

Piniel Alphayo Kambey1, 2, Wen-Ya Liu1, Jiao Wu1, Bakwatanisa Bosco3, Iqra Nadeem1, Kouminin Kanwore1, Dian-Shuai Gao1, *   

  1. 1Xuzhou Key Laboratory of Neurobiology, Department of Neurobiology and Anatomy, Xuzhou Medical University, Xuzhou, Jiangsu Province, China; 2Organization of African Academic Doctors, Nairobi, Kenya; 3Department of Bioinformatics, Nanjing Medical University, Nanjing, Jiangsu Province, China
  • Online:2023-09-15 Published:2023-03-07
  • Contact: Dian-Shuai Gao, MD, PhD, gds@xzhmu.edu.cn.
  • Supported by:
    The study was supported by the National Natural Science Foundation of China, No. 81971006 (to DSG).

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摘要：

既往研究发现帕金森病中核受体相关因子1存在缺失现象，而核受体相关因子1可参与多巴胺能神经元的发育分化、存活和变性，但是其对帕金森病的作用机制尚不明确。为此，首先通过联用液相色谱与串联质谱确定核受体相关因子1缺陷的影响，发现在核受体相关因子1缺陷的多巴胺能神经元中有231种基因处于高表达，且其中14种基因与京都百科全书上的帕金森病通路有关。进一步对慢病毒介导特异性短发夹RNA敲除核受体相关因子1小鼠模型进行单细胞核RNA测序，以探索核受体相关因子1缺失如何引起多巴胺能神经元的衰退并导致帕金森病的发生。结果可见小鼠大脑黑质中存在高调控基因及细胞间的异质性，其中显著相关性体现在主要组织相容性II复合体分子。Cd74，H2-Ab1，H2-Aa，H2-Eb1，Lyz2，Mrc1，Slc6a3，Slc47a1，Ms4a4b和Ptprc2是前10种最显著的差异表达基因。随后的免疫荧光染色结果显示，慢病毒介导特异性短发夹RNA敲除核受体相关因子1后，小鼠脑组织中Cd74免疫阳性细胞数量明显增加。最后以6-羟基多巴胺诱导的帕金森病小鼠模型进行验证，确认其中脑中Cd74基因表达增加。综上得出，核受体相关因子1缺失后，与帕金森病通路相关的基因编码蛋白Cd74表达升高，进而导致帕金森病多巴胺能神经元破坏，这将为帕金森病的治疗提供潜在的干预靶点。

http://orcid.org/0000-0001-8567-0238 (Dian-Shuai Gao)

关键词: 帕金森病, 6-羟基多巴胺, 多巴胺神经元, 酪氨酸羟化酶, 多巴胺转运体, 核受体相关因子1, 黑质, 蛋白质组学分析, 单核RNA测序, Seurat聚类

Abstract: Previous studies have found that deficiency in nuclear receptor-related factor 1 (Nurr1), which participates in the development, differentiation, survival, and degeneration of dopaminergic neurons, is associated with Parkinson’s disease, but the mechanism of action is perplexing. Here, we first ascertained the repercussion of knocking down Nurr1 by performing liquid chromatography coupled with tandem mass spectrometry. We found that 231 genes were highly expressed in dopaminergic neurons with Nurr1 deficiency, 14 of which were linked to the Parkinson’s disease pathway based on Kyoto Encyclopedia of Genes and Genomes analysis. To better understand how Nurr1 deficiency autonomously invokes the decline of dopaminergic neurons and elicits Parkinson’s disease symptoms, we performed single-nuclei RNA sequencing in a Nurr1 LV-shRNA mouse model. The results revealed cellular heterogeneity in the substantia nigra and a number of activated genes, the preponderance of which encode components of the major histocompatibility II complex. Cd74, H2-Ab1, H2-Aa, H2-Eb1, Lyz2, Mrc1, Slc6a3, Slc47a1, Ms4a4b, and Ptprc2 were the top 10 differentially expressed genes. Immunofluorescence staining showed that, after Nurr1 knockdown, the number of CD74-immunoreactive cells in mouse brain tissue was markedly increased. In addition, Cd74 expression was increased in a mouse model of Parkinson’s disease induced by treatment with 6-hydroxydopamine. Taken together, our results suggest that Nurr1 deficiency results in an increase in Cd74 expression, thereby leading to the destruction of dopaminergic neurons. These findings provide a potential therapeutic target for the treatment of Parkinson’s disease.

Key words: 6-hydroxydopamine, dopaminergic neurons, dopamine transporter, nuclear receptor-related factor 1, Parkinson’s disease, proteomics analysis, Seurat clustering, single-nuclei RNA sequencing, substantia nigra, tyrosine hydroxylase