DataSheet_1_Primitive genotypic characteristics in umbilical cord neutrophils identified by single-cell transcriptome profiling and functional prediction.docx

The function and heterogeneity of neutrophils in neonatal umbilical cord blood (UCB) have not been characterized. In this study, we analyzed the neutrophils in UCB and healthy adults using single-cell RNA sequencing analysis for the first time. We found that neutrophils divided into six subpopulations (G2, G3, G4, G5a, G5b, and G5c) with different marker genes and different functions under homeostasis. Compared with healthy adults, neutrophils of UCB were more naïve and have more obvious degranulation and activation functions. Moreover, we found significant differences in the amount and function of G5b cells between healthy adults and UCB. The amount of G5b group in UCB was lower, but it has more degranulation, secretion and activation functions. In addition, we noted a new subset of G5c labeled by CD52, which almost did not exist in UCB. Besides, its differential genes were enriched in terms such as protein synthesis and mRNA transcription. Furthermore, uncharacteristic transcription factors ZNF-276, ZNF-319 and ZNF-354A were identified in our study. In summary, we first examined the heterogeneity and functional diversity of neutrophils in UCB, and these data provided new insights into the mechanism of neutrophil-mediated diseases of neonates and the wider use of neutrophils in UCB.

新生儿脐带血（Umbilical Cord Blood，UCB）中性粒细胞的功能与异质性尚未得到系统阐明。本研究首次采用单细胞RNA测序技术，对UCB及健康成人外周血中的中性粒细胞展开分析。研究发现，在生理稳态下，中性粒细胞可分为G2、G3、G4、G5a、G5b及G5c共6个亚群，各亚群具备独特的标志物基因与功能特征。与健康成人相比，UCB来源的中性粒细胞表型更为未成熟，且脱颗粒与激活功能更为显著。此外，本研究发现健康成人与UCB中的G5b亚群在细胞数量与功能上均存在显著差异：UCB中G5b亚群的占比更低，但该亚群具备更强的脱颗粒、分泌及激活功能。除此之外，本研究还发现了一个由CD52标记的全新G5c亚群，该亚群在UCB中几乎不存在，其差异表达基因主要富集于蛋白质合成、mRNA转录等生物学过程。本研究同时鉴定出了ZNF-276、ZNF-319及ZNF-354A这三个尚未被深入表征的转录因子。综上，本研究首次系统解析了UCB中性粒细胞的异质性与功能多样性，相关数据为阐明新生儿中性粒细胞介导的疾病机制以及拓展UCB中性粒细胞的临床应用潜力提供了全新的研究视角。