Myeloid PDLIM2 repression as a common causal mechanism of lung disease pathogenesis and susceptibility to infection

Selective deletion of PDLIM2 in myeloid cells rendered mice more vulnerable to lung injury and mortality after LPS intratracheal instillation, which was associated with the exacerbated activation of pro-inflammation signaling pathways and the more diminished activation of anti-inflammation signaling pathways in the lung, and particularly, in lung macrophages and neutrophils. Overall design: The experiments started with WT and Pdlim2 myeloid KO (mKO) mice of similar age. Each group was further divided into PBS and LPS-treated ones where treatment was administered through intratracheal injection. Lungs from these 4 groups of mice were dissected 48hr post treatment and were then dissociated into single cell suspension. scRNAseq was done with the combination of 10x genomics single cell 5' kit and customized sample multiplexing using TotalSeq CD45 tag antibody.髓系细胞中PDLIM2的特异性敲除，可使小鼠在经气管内滴注脂多糖（LPS）后更易发生肺损伤且死亡率升高，该现象与肺部（尤其是肺巨噬细胞与中性粒细胞）中促炎信号通路的激活加剧、抗炎信号通路的激活进一步减弱显著相关。 实验设计：本实验以同龄野生型（Wild Type, WT）和髓系细胞特异性敲除Pdlim2（mKO）小鼠为研究对象，每组进一步分为磷酸盐缓冲液（PBS）处理组与LPS处理组，给药方式均为气管内注射。给药48小时后摘取四组小鼠的肺部组织，解离为单细胞悬液；采用10x Genomics单细胞5'建库试剂盒结合使用TotalSeq CD45标签抗体的定制化样本多重标记方案，完成单细胞RNA测序（scRNAseq）。