ASXL1 mutation correction by CRISPR/Cas9 restores gene function in leukemia cells and increases survival in mouse xenografts

Recurrent somatic mutations of the epigenetic modifier and tumor suppressor ASXL1 are common in myeloid malignancies, including chronic myeloid leukemia (CML), and are associated with poor clinical outcome. CRISPR/Cas9 has recently emerged as a powerful and versatile genome editing tool for genome e...

Full description

Saved in:

Bibliographic Details
Published in:	Oncotarget 2015-12, Vol.6 (42), p.44061-44071
Main Authors:	Valletta, Simona, Dolatshad, Hamid, Bartenstein, Matthias, Yip, Bon Ham, Bello, Erica, Gordon, Shanisha, Yu, Yiting, Shaw, Jacqueline, Roy, Swagata, Scifo, Laura, Schuh, Anna, Pellagatti, Andrea, Fulga, Tudor A, Verma, Amit, Boultwood, Jacqueline
Format:	Article
Language:	eng
Subjects:	Animals Base Sequence Cell Differentiation Cell Line, Tumor Cell Proliferation Clustered Regularly Interspaced Short Palindromic Repeats Codon, Nonsense CRISPR-Associated Proteins - genetics CRISPR-Associated Proteins - metabolism CRISPR-Cas Systems Gene Expression Regulation, Leukemic Genetic Predisposition to Disease Heterografts Homozygote Humans Interleukin Receptor Common gamma Subunit - deficiency Interleukin Receptor Common gamma Subunit - genetics Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics Leukemia, Myelogenous, Chronic, BCR-ABL Positive - metabolism Leukemia, Myelogenous, Chronic, BCR-ABL Positive - pathology Mice, Inbred NOD Mice, Knockout Mice, SCID Molecular Sequence Data Neoplasm Transplantation Phenotype Polycomb Repressive Complex 2 - genetics Polycomb Repressive Complex 2 - metabolism Priority Research Paper Repressor Proteins - genetics Repressor Proteins - metabolism Time Factors Tumor Suppressor Proteins - genetics Tumor Suppressor Proteins - metabolism Ubiquitin Thiolesterase - genetics Ubiquitin Thiolesterase - metabolism
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Recurrent somatic mutations of the epigenetic modifier and tumor suppressor ASXL1 are common in myeloid malignancies, including chronic myeloid leukemia (CML), and are associated with poor clinical outcome. CRISPR/Cas9 has recently emerged as a powerful and versatile genome editing tool for genome engineering in various species. We have used the CRISPR/Cas9 system to correct the ASXL1 homozygous nonsense mutation present in the CML cell line KBM5, which lacks ASXL1 protein expression. CRISPR/Cas9-mediated ASXL1 homozygous correction resulted in protein re-expression with restored normal function, including down-regulation of Polycomb repressive complex 2 target genes. Significantly reduced cell growth and increased myeloid differentiation were observed in ASXL1 mutation-corrected cells, providing new insights into the role of ASXL1 in human myeloid cell differentiation. Mice xenografted with mutation-corrected KBM5 cells showed significantly longer survival than uncorrected xenografts. These results show that the sole correction of a driver mutation in leukemia cells increases survival in vivo in mice. This study provides proof-of-concept for driver gene mutation correction via CRISPR/Cas9 technology in human leukemia cells and presents a strategy to illuminate the impact of oncogenic mutations on cellular function and survival.
ISSN:	1949-2553 1949-2553