单细胞分辨率下出生后猪肝脏的动态发育  

Single-cell dynamics of liver development in postnatal pigs

作　　者：饶琳 蔡丽萍 黄路生 Lin Rao;Liping Cai;Lusheng Huang(National Key Laboratory for Swine Genetic Improvement and Germplasm Innovation,Ministry of Science and Technology of China,Jiangxi Agricultural University,Nanchang 330045,China)

机构地区：[1]National Key Laboratory for Swine Genetic Improvement and Germplasm Innovation,Ministry of Science and Technology of China,Jiangxi Agricultural University,Nanchang 330045,China

出　　处：《Science Bulletin》2023年第21期2583-2597,M0005,共16页科学通报（英文版）

基　　金：supported by the National Natural Science Foundation of China(31790410);the National Pig Industry Technology system(CARS-35)。

摘　　要：肝脏作为代谢和免疫的重要器官,对其在出生后单细胞分辨率下的发育过程了解甚少.为此,本项目综合应用单核转录组以及单细胞转录组测序技术研究了猪出生后4个重要发育阶段(30,42,150,730天)肝脏的84824个细胞,鉴定到23种细胞类型,包括3种稀有细胞类型:浆细胞样树突状细胞、CAVIN3+IGF2+内皮细胞以及EBF1+成纤维细胞,并通过多重荧光免疫组化实验进行了验证.发育轨迹和基因调控网络分析揭示了33个与肝细胞发育和功能相关的转录因子编码基因,包括调节肝脏代谢的NFIL3.本文表征了肝脏内皮细胞的时空异质性,鉴定并验证了亮氨酸拉链蛋白2(LUZP2)是新的成年肝窦内皮细胞特异的转录因子.发现淋巴系免疫细胞(NK和T细胞)从出生后30天开始即主导了猪肝脏免疫系统.作者进一步鉴定到一簇肝脏组织驻留的NK细胞,该簇细胞呈现出病毒防御功能且在730天仍保持增殖特征.跨物种比较分析表明,与小鼠相比,猪肝脏驻留的NK细胞与人类表现出更保守的转录因子表达模式.本研究构建了迄今最为全面的出生后肝脏单细胞发育图谱并且阐释了其4个年龄阶段代谢和免疫的变化.The postnatal development of the liver,an essential organ for metabolism and immunity,remains poorly characterized at the single-cell resolution.Here,we generated single-nucleus and single-cell transcriptomes of 84,824 pig liver cells at four postnatal time points:day 30,42,150,and 730.We uncovered 23 cell types,including three rare cell types:plasmacytoid dendritic cells,CAVIN3+IGF2+endothelial cells,and EBF1+fibroblasts.The latter two were verified by multiplex immunohistochemistry.Trajectory and gene regulatory analyses revealed 33 genes that encode transcription factors associated with hepatocyte development and function,including NFIL3 involved in regulating hepatic metabolism.We characterized the spatiotemporal heterogeneity of liver endothelial cells,identified and validated leucine zipper protein 2(LUZP2)as a novel adult liver sinusoidal endothelial cell-specific transcription factor.Lymphoid cells(NK and T cells)governed the immune system of the pig liver since day 30.Furthermore,we identified a cluster of tissue-resident NK cells,which displayed virus defense functions,maintained proliferative features at day 730,and manifested a higher conservative transcription factor expression pattern in humans than in mouse liver.Our study presents the most comprehensive postnatal liver development single-cell atlas and demonstrates the metabolic and immune changes across the four age stages.

关 键 词：肝窦内皮细胞 淋巴系 基因调控网络 细胞类型 成纤维细胞 猪肝脏 免疫细胞 病毒防御 

分 类 号：R575[医药卫生—消化系统]