Decreased bone marrow regulatory innate lymphoid cells show a distinctive miRNA profiling in aplastic anemia

A new regulatory subpopulation of innate lymphoid cells (ILCs), regulatory innate lymphoid cells (ILCregs), has been identified with both innate lymphoid cells and regulatory cells characteristics. The purpose of this study is to explore ILCregs and its associated miRNAs in patients with aplastic anemia (AA) by evaluating ILCregs frequency, associated miRNA quantification, and their significance. Using 4 color combinations of surface and intracellular antibody staining, the CD45+Lin−CD127+IL-10+ ILCregs from 30 healthy donors and 30 patients newly diagnosed with AA were measured by flow cytometry. Bone marrow cells were studied by next-generation sequence miRNAs quantification. Our results showed that the frequency of ILCregs in bone marrow cells from healthy donors (HD) and AA patients were 0.703 ± 0.941 and 0.171 ± 0.233%, respectively. The frequencies of ILCregs in AA patients were significantly lower than that in HD (<i>p</i> &lt;0.05). miRNA detection results showed different expression patterns in the AA patient group comparing with HD. Comparing with HD, there were 52 miRNAs up-regulated and 130 miRNAs down-regulated from AA patients. Analysis of miRNAs from ILCregs associated genes demonstrated different miRNAs expression patterns between HD and AA patient. The present study demonstrated the deficiency of ILCregs and differential expression pattern of ILCregs gene-related miRNA in patients with AA. Further studies need to be done to explore the clinical significance of ILCregs and related miRNAs in patients with AA with large samples size and clinical follow-up.

本研究成功鉴定出一类兼具固有淋巴细胞（innate lymphoid cells, ILCs）与调节性细胞双重特性的新型调节性固有淋巴细胞亚群（regulatory innate lymphoid cells, ILCregs）。本研究旨在通过检测再生障碍性贫血（aplastic anemia, AA）患者体内调节性固有淋巴细胞的频数、相关微小RNA（microRNA, miRNAs）的表达水平及其临床意义，探索该亚群及其相关miRNAs在AA中的作用。本研究采用4种表面及胞内抗体染色组合方案，通过流式细胞术（flow cytometry）对30名健康志愿者（healthy donors, HD）及30例初诊AA患者的CD45+Lin⁻CD127+IL-10+调节性固有淋巴细胞进行定量检测；同时通过下一代测序技术（next-generation sequencing）对骨髓细胞中的miRNAs进行定量分析。结果显示，健康志愿者骨髓中调节性固有淋巴细胞的占比为0.703±0.941%，AA患者则为0.171±0.233%；AA患者骨髓中调节性固有淋巴细胞的频数显著低于健康志愿者（P<0.05）。miRNA检测结果显示，AA患者组与健康志愿者组的miRNA表达谱存在显著差异：与健康志愿者相比，AA患者体内共有52种miRNAs表达上调、130种miRNAs表达下调。通过分析与调节性固有淋巴细胞相关基因关联的miRNAs，发现健康志愿者与AA患者间的miRNA表达模式亦存在显著差异。本研究证实，AA患者体内存在调节性固有淋巴细胞数量缺乏的情况，且其调节性固有淋巴细胞相关基因的miRNA表达模式存在异常。后续仍需开展大样本量及临床随访相关研究，以进一步阐明调节性固有淋巴细胞及其相关miRNAs在AA患者中的临床意义。