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Targeting DDX3X eliminates leukemia stem cells in chronic myeloid leukemia by blocking NT5DC2 mRNA translation

Oncogene volume 44, pages 241–254 (2025)Cite this article

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Abstract

Tyrosine kinase inhibitors (TKIs) are highly effective in the treatment of patients with chronic myeloid leukemia (CML), but fail to eliminate leukemia stem cells (LSCs), which can lead to disease relapse or progression. It is urgently need to identify the regulators specifically driving LSCs. In this study, we identified DEAD-box helicase 3 X-linked (DDX3X), a ubiquitously expressed RNA helicase, as a critical regulator for CML LSCs by using patient samples and BCR-ABL-driven CML mouse model. We found that DDX3X enhanced the survival, serially plating and long-term engraftment abilities of human primary CML CD34+ cells. Inhibition of DDX3X reduced leukemia burden, eradicated LSCs and extended the survival of CML mice. Mechanistically, we uncovered that DDX3X protein bound to 5′-Nucleotidase Domain Containing 2 (NT5DC2) mRNA and promoted its translation in CML cells. NT5DC2 was a functional mediator in DDX3X regulation of LSCs. Collectively, our findings provide new evidence for RNA helicase facilitating the translation of specific mRNA in LSCs. Targeting DDX3X may represent a promising therapeutic strategy for eradication of LSCs in CML patients.

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Fig. 1: Targeting DDX3X inhibits the survival and serially plating features of human CML CD34+ cells.
Fig. 2: DDX3X inhibitor RK-33 suppresses the long-term engraftment ability of human CML CD34+ cells in NCG mice.
Fig. 3: RK-33 prolongs the survival of CML mice and eliminates LSCs.
Fig. 4: Ddx3x knockdown eliminates LSCs and inhibits the development of CML mice.
Fig. 5: DDX3X promotes the mRNA translation of NT5DC2 in CML cells.
Fig. 6: NT5DC2 knockdown induces apoptosis and inhibits the serially plating ability of human CML CD34+ cells.
Fig. 7: Nt5dc2 knockdown eliminates LSCs and inhibits the development of CML mice.
Fig. 8: Restoration of NT5DC2 rescues the Ddx3x knockdown-mediated elimination of LSCs in CML mice.

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Data availability

All data in this study are available in the main text or the supplementary information.

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Acknowledgements

This study was supported by grants from the National Natural Science Foundation of China (No. 82373897 to YJ), the Guangdong Basic and Applied Basic Research Foundation (No. 2024A1515010138 to YJ), Basic and Applied Basic Research Project of Guangzhou Basic Research Program (Nos. 2024A04J3325 and 2024A03J0451 to CL).

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Author notes

  1. These authors contributed equally: Chen Duan, Xiaoying Lin, Waiyi Zou, Qi He.

Authors and Affiliations

  1. State Key Laboratory of Bioactive Molecules and Druggability Assessment, Jinan University, Guangzhou, China

    Chen Duan, Xiaoying Lin, Qi He, Fen Wei, Chang Liu & Yanli Jin

  2. Institute of Tumor Pharmacology, College of Pharmacy, Jinan University, Guangzhou, China

    Chen Duan, Xiaoying Lin, Qi He, Fen Wei, Chang Liu & Yanli Jin

  3. Department of Hematology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China

    Waiyi Zou

  4. State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China

    Jingxuan Pan

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Contributions

Chen Duan, Xiaoying Lin, Qi He, Chang Liu, and Yanli Jin designed the research. Chen Duan, Xiaoying Lin, Qi He, and Fen Wei performed the experiments, analyzed, and interpreted the data. Waiyi Zou provided CML patient samples and information. Chang Liu and Yanli Jin advised on experiments and provided reagents. Chang Liu and Yanli Jin wrote the manuscript. Jingxuan Pan and Yanli Jin supervised the entire study. All authors reviewed the manuscript.

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Correspondence to Jingxuan Pan, Chang Liu or Yanli Jin.

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The authors declare no competing interests.

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All methods were performed in accordance with the relevant guidelines and regulations. This study involving patient samples was approved by the Jinan University Ethics Committee (JNUKY-2023-0063). Written informed consent was obtained from all CML patients. The animal experiments were performed with the approval of the Institutional Animal Care and Use Committee of South China University of Technology (2024108).

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Duan, C., Lin, X., Zou, W. et al. Targeting DDX3X eliminates leukemia stem cells in chronic myeloid leukemia by blocking NT5DC2 mRNA translation. Oncogene 44, 241–254 (2025). https://doi.org/10.1038/s41388-024-03215-w

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