The single cell RNA seq of pulmonary alveolar type II (AT2) cells. The single cell RNA seq of pulmonary alveolar type II (AT2) cells

The pulmonary alveolar epithelium mainly composed of two types of epithelial cells: alveolar type I (AT1) and type II (AT2) cells. AT2 cells are the alveolar stem cells, and can differentiate into AT1 cells post-pneumonectomy (PNX). Here, we found that, compared with control mice (Sftpc-CreER; Cdc42flox/+; Rosa26-mTmG) at post-PNX day 21, Cdc42 AT2 null mice (Sftpc-CreER; Cdc42flox/-; Rosa26-mTmG) at post-PNX day 21 undergone fibrotic change. By using 10X genomics “Chromium Single Cell” technology, we performed single-cell RNA-seq analyses of AT2 cells of sham treated control mice (C0), AT2 cells of control mice at post PNX day 21 (C21) , AT2 cells of sham treated Cdc42 AT2 null mice (N0), and AT2 cells of Cdc42 AT2 null mice at post PNX day 21 (N21). The study identified a specific gene signature in AT2 cells of Cdc42 AT2 null mice at post PNX day 21 which is related to the fibrosis phenotype of Cdc42 AT2 null mice. Overall design: We performed 10X genomics single-cell RNA-seq of AT2 cells of sham treated control mice (C0), AT2 cells of control mice at post PNX day 21 (C21) , AT2 cells of sham treated Cdc42 AT2 null mice ( N0) and AT2 cells of Cdc42 AT2 null mice at post PNX day 21 (N21).

肺泡上皮主要由两类上皮细胞构成：肺泡I型（AT1）细胞与肺泡II型（AT2）细胞。AT2细胞作为肺泡干细胞，可在肺切除术（pneumonectomy, PNX）后分化为AT1细胞。本研究发现，相较于肺切除术后第21天的对照小鼠（Sftpc-CreER; Cdc42flox/+; Rosa26-mTmG），同时间点的Cdc42 AT2细胞特异性敲除小鼠（Sftpc-CreER; Cdc42flox/-; Rosa26-mTmG）出现了纤维化改变。本研究借助10×基因组学"Chromium Single Cell"技术，对四组小鼠的AT2细胞开展了单细胞RNA测序（single-cell RNA-seq）分析，四组样本分别为：假手术处理的对照小鼠AT2细胞（C0）、肺切除术后第21天的对照小鼠AT2细胞（C21）、假手术处理的Cdc42 AT2细胞特异性敲除小鼠AT2细胞（N0），以及肺切除术后第21天的Cdc42 AT2细胞特异性敲除小鼠AT2细胞（N21）。本研究鉴定得到，肺切除术后第21天的Cdc42 AT2细胞特异性敲除小鼠的AT2细胞中存在与该小鼠纤维化表型相关的特异性基因特征。整体实验设计：我们对上述四组样本的AT2细胞实施了10×基因组学单细胞RNA测序，样本分别为：假手术处理的对照小鼠AT2细胞（C0）、肺切除术后第21天的对照小鼠AT2细胞（C21）、假手术处理的Cdc42 AT2细胞特异性敲除小鼠AT2细胞（N0），以及肺切除术后第21天的Cdc42 AT2细胞特异性敲除小鼠AT2细胞（N21）。