Continuous genetic monitoring of transient mesenchymal gene activities in distal tubule and collecting duct epithelial cells during renal fibrosis

Author:

Xu Zihang123,Zhang Shaotong1,Han Tingting1,Cai Letong1,Zhong Simin1,Yang Xiaojie1,Zhang Shaohua4,Li Yan4,Liu Kuo5,Zhou Bin4567,Tian Xueying23ORCID

Affiliation:

1. Key Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, College of Life Science and Technology Jinan University Guangzhou China

2. Obstetrics and Gynecology Hospital, Institute of Reproduction and Development, Shanghai Medical College Fudan University Shanghai China

3. Shanghai Key Laboratory of Reproduction and Development Shanghai China

4. State Key Laboratory of Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai Institute of Biochemistry and Cell Biology University of Chinese Academy of Sciences Shanghai China

5. Key Laboratory of Systems Health Science of Zhejiang Province, School of Life Science, Hangzhou Institute for Advanced Study University of Chinese Academy of Sciences Hangzhou China

6. School of Life Science and Technology ShanghaiTech University Shanghai China

7. New Cornerstone Science Laboratory Shenzhen China

Abstract

AbstractEpithelial cells (ECs) have been proposed to contribute to myofibroblasts or fibroblasts through epithelial‐mesenchymal transition (EMT) during renal fibrosis. However, since EMT may occur dynamically, transiently, and reversibly during kidney fibrosis, conventional lineage tracing based on Cre‐loxP recombination in renal ECs could hardly capture the transient EMT activity, yielding inconsistent results. Moreover, previous EMT research has primarily focused on renal proximal tubule ECs, with few reports of distal tubules and collecting ducts. Here, we generated dual recombinases‐mediated genetic lineage tracing systems for continuous monitoring of transient mesenchymal gene expression in E‐cadherin+ and EpCAM+ ECs of distal tubules and collecting ducts during renal fibrosis. Activation of key EMT‐inducing transcription factor (EMT‐TF) Zeb1 and mesenchymal markers αSMA, vimentin, and N‐cadherin, were investigated following unilateral ureteral obstruction (UUO). Our data revealed that E‐cadherin+ and EpCAM+ ECs did not transdifferentiate into myofibroblasts, nor transiently expressed these mesenchymal genes during renal fibrosis. In contrast, in vitro a large amount of cultured renal ECs upregulated mesenchymal genes in response to TGF‐β, a major inducer of EMT.

Publisher

Wiley

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