Retinoic acid and arsenic trioxide induce lasting differentiation and demethylation of target genes in APL cells

Thomas T. Huynh; Mohammad Sultan; Dejan Vidovic; Cheryl A. Dean; Brianne M. Cruickshank; Kristen Lee; Chao Yu Loung; Ryan W. Holloway; David W. Hoskin; David M. Waisman; Ian C.G. Weaver; Paola Marcato

doi:10.1038/s41598-019-45982-7

Retinoic acid and arsenic trioxide induce lasting differentiation and demethylation of target genes in APL cells

Thomas T. Huynh, Mohammad Sultan, Dejan Vidovic, Cheryl A. Dean, Brianne M. Cruickshank, Kristen Lee, Chao Yu Loung, Ryan W. Holloway, David W. Hoskin, David M. Waisman, Ian C.G. Weaver, Paola Marcato

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Résumé

Acute promyelocytic leukemia (APL) is characterized by arrested differentiation of promyelocytes. Patients treated with all-trans retinoic acid (ATRA) alone experience relapse, while patients treated with ATRA and arsenic trioxide (ATO) are often relapse-free. This suggests sustained changes have been elicited by the combination therapy. To understand the lasting effects of the combination therapy, we compared the effects of ATRA and ATO on NB4 and ATRA-resistant NB4-MR2 APL cells during treatment versus post treatment termination. After treatment termination, NB4 cells treated with ATRA or ATO reverted to non-differentiated cells, while combination-treated cells remained terminally differentiated. This effect was diminished in NB4-MR2 cells. This suggests combination treatment induced more permanent changes. Combination treatment induced higher expression of target genes (e.g., transglutaminase 2 and retinoic acid receptor beta), which in NB4 cells was sustained post treatment termination. To determine whether sustained epigenetic changes were responsible, we quantified the enrichment of histone modifications by chromatin immunoprecipitation, and CpG methylation by bisulfite-pyrosequencing. While ATRA and combination treatment induced similar histone acetylation enrichment, combination treatment induced greater demethylation of target genes, which was sustained. Therefore, sustained demethylation of target genes by ATRA and ATO combination treatment is associated with lasting differentiation and gene expression changes.

Langue d'origine	English
Numéro d'article	9414
Journal	Scientific Reports
Volume	9
Numéro de publication	1
DOI	https://doi.org/10.1038/s41598-019-45982-7
Statut de publication	Published - déc. 1 2019

Note bibliographique

Funding Information:
Support was provided by grant funding to PM from the Canadian Institutes of Health Research (CIHR, MOP-130304 and PJT-162313), to ICGW from the Natural Sciences and Engineering Research Council of Canada (NSERC, RGPIN-2013-436204) and to THH and KL from the Beatrice Hunter Cancer Research Institute (BHCRI, collaborative grant). TTH was supported by a studentship from the BCHRI, the Canadian Imperial Bank of Commerce and a Nova Scotia Research and Innovation Graduate scholarship. DV, MS, and BMC were supported by a Nova Scotia Research and Innovation Graduate scholarship and Killam Laureate scholarship. The flow cytometry data was obtained by accessing Dalhousie University’s Faculty of Medicine Flow Cytometry core facility.

Publisher Copyright:
© 2019, The Author(s).

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Huynh, T. T., Sultan, M., Vidovic, D., Dean, C. A., Cruickshank, B. M., Lee, K., Loung, C. Y., Holloway, R. W., Hoskin, D. W., Waisman, D. M., Weaver, I. C. G., & Marcato, P. (2019). Retinoic acid and arsenic trioxide induce lasting differentiation and demethylation of target genes in APL cells. Scientific Reports, 9(1), Article 9414. https://doi.org/10.1038/s41598-019-45982-7

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title = "Retinoic acid and arsenic trioxide induce lasting differentiation and demethylation of target genes in APL cells",

abstract = "Acute promyelocytic leukemia (APL) is characterized by arrested differentiation of promyelocytes. Patients treated with all-trans retinoic acid (ATRA) alone experience relapse, while patients treated with ATRA and arsenic trioxide (ATO) are often relapse-free. This suggests sustained changes have been elicited by the combination therapy. To understand the lasting effects of the combination therapy, we compared the effects of ATRA and ATO on NB4 and ATRA-resistant NB4-MR2 APL cells during treatment versus post treatment termination. After treatment termination, NB4 cells treated with ATRA or ATO reverted to non-differentiated cells, while combination-treated cells remained terminally differentiated. This effect was diminished in NB4-MR2 cells. This suggests combination treatment induced more permanent changes. Combination treatment induced higher expression of target genes (e.g., transglutaminase 2 and retinoic acid receptor beta), which in NB4 cells was sustained post treatment termination. To determine whether sustained epigenetic changes were responsible, we quantified the enrichment of histone modifications by chromatin immunoprecipitation, and CpG methylation by bisulfite-pyrosequencing. While ATRA and combination treatment induced similar histone acetylation enrichment, combination treatment induced greater demethylation of target genes, which was sustained. Therefore, sustained demethylation of target genes by ATRA and ATO combination treatment is associated with lasting differentiation and gene expression changes.",

author = "Huynh, {Thomas T.} and Mohammad Sultan and Dejan Vidovic and Dean, {Cheryl A.} and Cruickshank, {Brianne M.} and Kristen Lee and Loung, {Chao Yu} and Holloway, {Ryan W.} and Hoskin, {David W.} and Waisman, {David M.} and Weaver, {Ian C.G.} and Paola Marcato",

note = "Funding Information: Support was provided by grant funding to PM from the Canadian Institutes of Health Research (CIHR, MOP-130304 and PJT-162313), to ICGW from the Natural Sciences and Engineering Research Council of Canada (NSERC, RGPIN-2013-436204) and to THH and KL from the Beatrice Hunter Cancer Research Institute (BHCRI, collaborative grant). TTH was supported by a studentship from the BCHRI, the Canadian Imperial Bank of Commerce and a Nova Scotia Research and Innovation Graduate scholarship. DV, MS, and BMC were supported by a Nova Scotia Research and Innovation Graduate scholarship and Killam Laureate scholarship. The flow cytometry data was obtained by accessing Dalhousie University{\textquoteright}s Faculty of Medicine Flow Cytometry core facility. Publisher Copyright: {\textcopyright} 2019, The Author(s).",

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AU - Huynh, Thomas T.

AU - Sultan, Mohammad

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AU - Dean, Cheryl A.

AU - Cruickshank, Brianne M.

AU - Lee, Kristen

AU - Loung, Chao Yu

AU - Holloway, Ryan W.

AU - Hoskin, David W.

AU - Waisman, David M.

AU - Weaver, Ian C.G.

AU - Marcato, Paola

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Retinoic acid and arsenic trioxide induce lasting differentiation and demethylation of target genes in APL cells

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