Yue Xinyi, Hu Qianwen, Cheng Haoyang, Su Gang, Diao Jiuzhou, Du Yansheng, Fu Pengrui, Wang Ping, Zhang Leisheng, Zhou Qingbo
Department of Neurology, Shandong public health clinical center, Shandong University, Jinan, 250100, China.
Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, 400038, China.
Sci Rep. 2025 Nov 28;15(1):42750. doi: 10.1038/s41598-025-26756-w.
Parkinson's disease (PD) is an intractable progressive neurodegenerative disease with poor prognosis in elder patients, which is largely attribute to the deficiency of ultimate pathogenesis. Longitudinal studies have indicated a pivotal role of natural killer (NK) cells, yet the systematic and detailed information of circulating NK cells in PD is largely unknowable. To explore the role of NK cells in PD, we isolated mononuclear cells from peripheral blood (PBMCs) by Ficoll-based density gradient centrifugation, and detected the content of total resident NK cells in healthy donors (HD-NKs) and PD patients (PD-NKs) and the concomitant subsets by flow cytometry (FCM) assay. Then, we took advantage of our well-established "3ILs"-based strategy for ex vivo NK cell expansion and activation, and followed by cellular viability and cytotoxicity assessment. By conducting RNA-sequencing (RNA-SEQ) and multifaceted bioinformatics analyses, we compared the transcriptomic signatures of expanded HD-NKs and PD-NKs. Compared to HD-NKs, PD-NKs showed increase in resident NK cells but minimal differences in expanded NK cells, together with diversity in the subpopulations of NK cells (CD16, NKG2D, NKp46). Interestingly, PD-NKs revealed a moderate higher percentage of apoptotic cells and cytotoxicity upon the co-cultured Nalm6 and U937 tumor cell lines. Despite with high conservations in gene expression pattern and genetic variations, PD-NKs revealed multifaceted diversity in gene set-associated immune response and metabolism. Overall, our data revealed the multidimensional biological and transcriptomic signatures of resident and expanded NK cells generated from peripheral blood of PD patients and HDs. Our findings would provide new references for the further development of NK cell-based biomarkers for PD diagnosis and novel immunotherapy for neurodegenerative diseases.
帕金森病（PD）是一种难以治愈的进行性神经退行性疾病，老年患者预后较差，这在很大程度上归因于最终发病机制的缺乏。纵向研究表明自然杀伤（NK）细胞起关键作用，但PD患者循环NK细胞的系统详细信息在很大程度上尚不清楚。为了探究NK细胞在PD中的作用，我们通过基于Ficoll的密度梯度离心从外周血中分离单核细胞（PBMCs），并通过流式细胞术（FCM）检测健康供体（HD-NKs）和PD患者（PD-NKs）中总驻留NK细胞的含量以及伴随的亚群。然后，我们利用成熟的基于“3ILs”的策略进行体外NK细胞扩增和激活，随后进行细胞活力和细胞毒性评估。通过进行RNA测序（RNA-SEQ）和多方面的生物信息学分析，我们比较了扩增后的HD-NKs和PD-NKs的转录组特征。与HD-NKs相比，PD-NKs的驻留NK细胞增加，但扩增后的NK细胞差异最小，同时NK细胞亚群（CD16、NKG2D、NKp46）存在多样性。有趣的是，PD-NKs在与Nalm6和U937肿瘤细胞系共培养时显示出适度较高比例的凋亡细胞和细胞毒性。尽管基因表达模式和基因变异具有高度保守性，但PD-NKs在基因集相关的免疫反应和代谢方面显示出多方面的多样性。总体而言，我们的数据揭示了PD患者和HDs外周血中驻留和扩增的NK细胞的多维生物学和转录组特征。我们的研究结果将为进一步开发基于NK细胞的PD诊断生物标志物和神经退行性疾病的新型免疫疗法提供新的参考。
