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

Simona Valletta, Hamid Dolatshad, Matthias Bartenstein, Bon Ham Yip, Erica Bello, Shanisha Gordon, Yiting Yu, Jacqueline Shaw, Swagata Roy, Laura Scifo, Anna Schuh, Andrea Pellagatti, Tudor A. Fulga, Amit K. Verma, Jacqueline Boultwood

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)44061-44071
Number of pages11
JournalOncotarget
Volume6
Issue number42
DOIs
StatePublished - 2015

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Heterografts
Cell Survival
Leukemia
Mutation
Genes
Myeloid Cells
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Polycomb Repressive Complex 2
Survival
Nonsense Codon
Epigenomics
Cell Differentiation
Neoplasms
Proteins
Down-Regulation
Genome
Technology
Cell Line
Growth

Keywords

  • ASXL1
  • Chronic myeloid leukemia
  • CRISPR
  • Mutation correction
  • Tumor suppressor

ASJC Scopus subject areas

  • Oncology

Cite this

Valletta, S., Dolatshad, H., Bartenstein, M., Yip, B. H., Bello, E., Gordon, S., ... Boultwood, J. (2015). ASXL1 mutation correction by CRISPR/Cas9 restores gene function in leukemia cells and increases survival in mouse xenografts. Oncotarget, 6(42), 44061-44071. https://doi.org/10.18632/oncotarget.6392

ASXL1 mutation correction by CRISPR/Cas9 restores gene function in leukemia cells and increases survival in mouse xenografts. / 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 K.; Boultwood, Jacqueline.

In: Oncotarget, Vol. 6, No. 42, 2015, p. 44061-44071.

Research output: Contribution to journalArticle

Valletta, S, Dolatshad, H, Bartenstein, M, Yip, BH, Bello, E, Gordon, S, Yu, Y, Shaw, J, Roy, S, Scifo, L, Schuh, A, Pellagatti, A, Fulga, TA, Verma, AK & Boultwood, J 2015, 'ASXL1 mutation correction by CRISPR/Cas9 restores gene function in leukemia cells and increases survival in mouse xenografts', Oncotarget, vol. 6, no. 42, pp. 44061-44071. https://doi.org/10.18632/oncotarget.6392
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 K. ; Boultwood, Jacqueline. / ASXL1 mutation correction by CRISPR/Cas9 restores gene function in leukemia cells and increases survival in mouse xenografts. In: Oncotarget. 2015 ; Vol. 6, No. 42. pp. 44061-44071.
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