Open Access

CRISPR/Cas9-mediated ASXL1 mutations in U937 cells disrupt myeloid differentiation

  • Authors:
    • Zhi-Jie Wu
    • Xin Zhao
    • Lauren G. Banaszak
    • Fernanda Gutierrez-Rodrigues
    • Keyvan Keyvanfar
    • Shou-Guo Gao
    • Diego Quinones Raffo
    • Sachiko Kajigaya
    • Neal S. Young
  • View Affiliations

  • Published online on: February 28, 2018     https://doi.org/10.3892/ijo.2018.4290
  • Pages: 1209-1223
  • Copyright: © Wu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Additional sex combs-like 1 (ASXL1) is a well‑known tumor suppressor gene and epigenetic modifier. ASXL1 mutations are frequent in myeloid malignances; these mutations are risk factors for the development of myelodysplasia and also appear as small clones during normal aging. ASXL1 appears to act as an epigenetic regulator of cell survival and myeloid differentiation; however, the molecular mechanisms underlying the malignant transformation of cells with ASXL1 mutations are not well defined. Using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) genome editing, heterozygous and homozygous ASXL1 mutations were introduced into human U937 leukemic cells. Comparable cell growth and cell cycle progression were observed between wild-type (WT) and ASXL1-mutated U937 cells. Drug-induced cytotoxicity, as measured by growth inhibition and apoptosis in the presence of the cell-cycle active agent 5-fluorouracil, was variable among the mutated clones but was not significantly different from WT cells. In addition, ASXL1-mutated cells exhibited defects in monocyte/macrophage differentiation. Transcriptome analysis revealed that ASXL1 mutations altered differentiation of U937 cells by disturbing genes involved in myeloid differentiation, including cytochrome B-245 β chain and C-type lectin domain family 5, member A. Dysregulation of numerous gene sets associated with cell death and survival were also observed in ASXL1-mutated cells. These data provide evidence regarding the underlying molecular mechanisms induced by mutated ASXL1 in leukemogenesis.
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April-2018
Volume 52 Issue 4

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Wu Z, Zhao X, Banaszak LG, Gutierrez-Rodrigues F, Keyvanfar K, Gao S, Quinones Raffo D, Kajigaya S and Young NS: CRISPR/Cas9-mediated ASXL1 mutations in U937 cells disrupt myeloid differentiation. Int J Oncol 52: 1209-1223, 2018.
APA
Wu, Z., Zhao, X., Banaszak, L.G., Gutierrez-Rodrigues, F., Keyvanfar, K., Gao, S. ... Young, N.S. (2018). CRISPR/Cas9-mediated ASXL1 mutations in U937 cells disrupt myeloid differentiation. International Journal of Oncology, 52, 1209-1223. https://doi.org/10.3892/ijo.2018.4290
MLA
Wu, Z., Zhao, X., Banaszak, L. G., Gutierrez-Rodrigues, F., Keyvanfar, K., Gao, S., Quinones Raffo, D., Kajigaya, S., Young, N. S."CRISPR/Cas9-mediated ASXL1 mutations in U937 cells disrupt myeloid differentiation". International Journal of Oncology 52.4 (2018): 1209-1223.
Chicago
Wu, Z., Zhao, X., Banaszak, L. G., Gutierrez-Rodrigues, F., Keyvanfar, K., Gao, S., Quinones Raffo, D., Kajigaya, S., Young, N. S."CRISPR/Cas9-mediated ASXL1 mutations in U937 cells disrupt myeloid differentiation". International Journal of Oncology 52, no. 4 (2018): 1209-1223. https://doi.org/10.3892/ijo.2018.4290