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Translational Therapeutics

Combination of a synthetic retinoid and a DNA demethylating agent induced differentiation of neuroblastoma through retinoic acid signal reprogramming

British Journal of Cancer volume 125, pages 1647–1656 (2021)Cite this article

Subjects

Abstract

Background

The CpG island methylator phenotype of neuroblastoma (NBL) is strongly associated with poor prognosis and can be targeted by 5-aza-2’-deoxycytidine (5-aza-dC). Differentiation therapy is a standard maintenance therapy for high-risk NBLs. However, the in vivo effect of tamibarotene, a synthetic retinoic acid, and the efficacy of its combination with 5-aza-dC have not been studied. Here, we conducted a preclinical study to assess the in vivo tamibarotene effect and the combination.

Methods

Treatment effects were analysed by in vitro cell growth and differentiation state and by in vivo xenograft suppression. Demethylated genes were analysed by DNA methylation microarrays and geneset enrichment.

Results

Tamibarotene monotherapy induced neural extension and upregulation of differentiation markers of NBL cells in vitro, and tumour regression without severe side effects in vivo. 5-Aza-dC monotherapy suppressed tumour growth both in vitro and in vivo, and induced demethylation of genes related to nervous system development and function. Pre-treatment with 5-aza-dC in vitro enhanced upregulation of differentiation markers and genes involved in retinoic acid signaling. Pre-treatment with 5-aza-dC in vivo significantly suppressed tumour growth and reduced the variation in tumour sizes.

Conclusions

Epigenetic drug-based differentiation therapy using 5-aza-dC and TBT is a promising strategy for refractory NBLs.

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Fig. 1: Effects of TBT on neuroblastoma differentiation and tumour growth.
Fig. 2: Effects of 5-aza-dC on tumour growth.
Fig. 3: Effects of 5-aza-dC on DNA methylation.
Fig. 4: Effects of the combination of 5-aza-dC and TBT on neuroblastoma differentiation.
Fig. 5: Effects of a combination of 5-aza-dC and TBT on tumour growth.

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Data availability

The datasets used in this study are available at the Gene Expression Omnibus (GEO) database (accession no. GSE180645, GSE180660 and GSE180929, https://www.ncbi.nlm.nih.gov/geo/).

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Acknowledgements

We thank Dr. H. Kawamoto for his critical discussion. We also thank Dr. T. Imai, Dr. N. Uchiya and Ms. Y. Shiotani, and the National Cancer Center Research Core Facility for their assistance in the analysis of xenografts.

Funding

The Core Facility was supported in part by National Cancer Center Research and Development Fund (2020-J-2). This study was supported by the AMED under Grant Number JP20ck0106421 to TU, NH, CN and JH, and JSPS KAKENHI Grant Number JP18H02704 to TU and NH.

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Authors and Affiliations

  1. Division of Epigenomics, National Cancer Center Research Institute, Tokyo, Japan

    Naoko Hattori, Kiyoshi Asada, Nozomu Miyajima, Akiko Mori, Yoko Nakanishi, Kana Kimura, Mika Wakabayashi, Hideyuki Takeshima & Toshikazu Ushijima

  2. Department of Pediatric Hematology and Oncology, Osaka City General Hospital, Osaka, Japan

    Chika Nitani & Junichi Hara

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Contributions

TU, NH, KA, CN and JH designed the study; NH, NM, AM, YN, KK, MW and HT performed most of the experiments; NH and TU wrote the manuscript; and all authors read and approved the manuscript.

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Correspondence to Naoko Hattori or Toshikazu Ushijima.

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All animal experiments were approved by the Committee of National Cancer Center and were conducted according to the institutional guidelines for animal care.

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Hattori, N., Asada, K., Miyajima, N. et al. Combination of a synthetic retinoid and a DNA demethylating agent induced differentiation of neuroblastoma through retinoic acid signal reprogramming. Br J Cancer 125, 1647–1656 (2021). https://doi.org/10.1038/s41416-021-01571-y

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