Single-cell Sequencing Reveals Variants in ARID1A, GPRC5A and MLL2 Driving Self-renewal of Human Bladder Cancer Stem Cells

Zhao Yang; Chong Li; Zusen Fan; Hongjie Liu; Xiaolong Zhang; Zhiming Cai; Liqin Xu; Jian Luo; Yi Huang; Luyun He; Chunxiao Liu; Song Wu

doi:10.1016/j.eururo.2016.06.025

Single-cell Sequencing Reveals Variants in ARID1A, GPRC5A and MLL2 Driving Self-renewal of Human Bladder Cancer Stem Cells

Eur Urol. 2017 Jan;71(1):8-12. doi: 10.1016/j.eururo.2016.06.025. Epub 2016 Jul 4.

Authors

Zhao Yang¹, Chong Li¹, Zusen Fan¹, Hongjie Liu², Xiaolong Zhang³, Zhiming Cai⁴, Liqin Xu⁵, Jian Luo⁵, Yi Huang³, Luyun He¹, Chunxiao Liu⁶, Song Wu⁷

Affiliations

¹ CAS Key Laboratory of Infection and Immunity, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
² BGI Education Centre, University of Chinese Academy of Sciences, Shenzhen 518083, China; College of Life Science, University of Chinese Academy of Sciences College, Beijing 100049, China.
³ Department of Urological Surgery, The Affiliated Luohu Hospital of Shenzhen University, Shenzhen University, Shenzhen 518000, China.
⁴ Department of Urological Surgery, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen 518000, China.
⁵ BGI-Shenzhen, Shenzhen 518000, China.
⁶ Department of Urology, Zhujiang Hospital of Southern Medical University, Guangzhou 510000, China.
⁷ Department of Urological Surgery, The Affiliated Luohu Hospital of Shenzhen University, Shenzhen University, Shenzhen 518000, China. Electronic address: wusong@szu.edu.cn.

PMID: 27387124
DOI: 10.1016/j.eururo.2016.06.025

Abstract

Cancer stem cells are considered responsible for many important aspects of tumors such as their self-renewal, tumor-initiating, drug-resistance and metastasis. However, the genetic basis and origination of human bladder cancer stem cells (BCSCs) remains unknown. Here, we conducted single-cell sequencing on 59 cells including BCSCs, bladder cancer non-stem cells (BCNSCs), bladder epithelial stem cells (BESCs) and bladder epithelial non-stem cells (BENSCs) from three bladder cancer (BC) specimens. Specifically, BCSCs demonstrate clonal homogeneity and suggest their origin from BESCs or BCNSCs through phylogenetic analysis. Moreover, 21 key altered genes were identified in BCSCs including six genes not previously described in BC (ETS1, GPRC5A, MKL1, PAWR, PITX2 and RGS9BP). Co-mutations of ARID1A, GPRC5A and MLL2 introduced by CRISPR/Cas9 significantly enhance the capabilities of self-renewal and tumor-initiating of BCNSCs. To our knowledge, our study first provides an overview of the genetic basis of human BCSCs with single-cell sequencing and demonstrates the biclonal origin of human BCSCs via evolution analysis.

Patient summary: Human bladder cancer stem cells show the high level of consistency and may derived from bladder epithelial stem cells or bladder cancer non-stem cells. Mutations of ARID1A, GPRC5A and MLL2 grant bladder cancer non-stem cells the capability of self-renewal.

Keywords: bladder cancer; bladder cancer stem cells; genetic alteration; self-renewal; single-cell sequencing; tumor evolution.

MeSH terms

DNA-Binding Proteins / genetics*
Epithelial Cells / metabolism
Epithelial Cells / physiology
Exome Sequencing
Humans
Mutation
Neoplasm Proteins / genetics*
Neoplastic Stem Cells / metabolism*
Neoplastic Stem Cells / physiology
Nuclear Proteins / genetics*
Polymorphism, Single Nucleotide
Receptors, G-Protein-Coupled / genetics*
Transcription Factors / genetics*
Urinary Bladder Neoplasms / genetics*
Urinary Bladder Neoplasms / physiopathology

Substances

ARID1A protein, human
DNA-Binding Proteins
GPRC5A protein, human
KMT2D protein, human
Neoplasm Proteins
Nuclear Proteins
Receptors, G-Protein-Coupled
Transcription Factors