Mapping the initial effects of carcinogen-induced oncogenic transformation in the mouse bladder

BBN is a bladder specific carcinogen that can cause invasive bladder cancer in mice possessing similar genetic alterations as seen in human cancer. We employed a single cell sequencing approach using 10x Genomics scRNAseq to investigate the cellular heterogeneity and molecular modifications induced by long term treatment with a carcinogen (BBN) on the bladder urothelium and sub-urothelium cells. Our data show infiltrations of B- and T-cells in the tumor and decrease of the several other urothelial populations. Overall design: 3 sample analysed: 2 non-treated controls and 1 BBN-treated. After 22 weeks of BBN treatment, the mucosa layer of mouse bladder was separated from detrusor and digested to prepare a cell suspension and single live cells were sorted using Fluorescence-activated cell sorting (FACS). Then scRNA-seq was performed to analyze the cells. The same was done for two age-matched non-treated mice.

BBN是一种膀胱特异性致癌物，可在携带有与人类癌症相似遗传改变的小鼠中诱发浸润性膀胱癌。本研究采用10x Genomics单细胞RNA测序（scRNAseq）技术，探究长期暴露于致癌物BBN后，小鼠膀胱尿路上皮及尿路上皮下细胞的细胞异质性与分子修饰变化。研究数据显示，肿瘤组织中存在B细胞与T细胞浸润，且其余多种尿路上皮细胞群的占比出现下降。

整体实验设计：共分析3组样本：2组未处理对照组与1组BBN处理组。BBN处理持续22周后，我们将小鼠膀胱黏膜层与逼尿肌分离，经消化制备细胞悬液，并通过荧光激活细胞分选（Fluorescence-activated cell sorting, FACS）分选存活的单个细胞。随后开展单细胞RNA测序以分析细胞样本。2只年龄匹配的未处理小鼠也采用相同流程开展实验。