Transcriptomic signature associated with education in uterine Natural Killer cells during pregnancy

Uterine natural killer (uNK) cells regulate vascular changes crucial for placental function and fetal growth. The functional capacity of uNK cells is shaped by their education status. To explore the mechanisms of uNK cell education, we isolated NKG2A+ and Ly49C/I+ educated uNK cells and compared their transcriptomes to uneducated NKG2A-Ly49C/I- uNK cells. At gestation days 9.510.5, pregnant mouse uteri were enzymatically and mechanically dissociated into single-cell suspensions. NK cells were isolated based on the expression of educating receptors Ly49C/I and NKG2A into three populations: uneducated (Ly49C/I-NKG2A-) and educated (Ly49C/I+NKG2A-, Ly49C/I-NKG2A+). Double-positive cells (Ly49C+/I+NKG2A) were excluded due to limited sample size. Bulk RNA-seq was performed to compare transcriptomic profiles between these populations, identifying differential gene expression associated with uNK cell education.子宫自然杀伤（Uterine natural killer, uNK）细胞可调控对胎盘功能与胎儿生长至关重要的血管重塑过程。uNK细胞的功能特性由其教育状态所决定。为探究uNK细胞教育的分子机制，本研究分离得到表达NKG2A与Ly49C/I的已教育型uNK细胞，并将其转录组与未教育型（NKG2A⁻Ly49C/I⁻）uNK细胞的转录组进行对比分析。本研究于妊娠第9.5至10.5天，通过酶解联合机械解离的方法将妊娠小鼠子宫制备为单细胞悬液。基于教育性受体Ly49C/I与NKG2A的表达水平，将NK细胞划分为三个亚群：未教育型（Ly49C/I⁻NKG2A⁻）以及两类已教育型（Ly49C/I⁺NKG2A⁻、Ly49C/I⁻NKG2A⁺）。由于样本量有限，双阳性细胞（Ly49C⁺/I⁺NKG2A⁺）被排除出本次研究。本研究通过批量RNA测序（bulk RNA-seq）对比上述各亚群的转录组特征，筛选出与uNK细胞教育相关的差异表达基因。