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ACUTE LYMPHOBLASTIC LEUKEMIA

TLE4 is a repressor of the oncogenic activity of TLX3 in T-cell acute lymphoblastic leukemia

Leukemia volume 39, pages 568–576 (2025)Cite this article

Abstract

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological disease originating from the malignant transformation of T-cell progenitors, caused by the accumulation of genetic aberrations. One-fifth of T-ALL patients are characterized by ectopic expression of the homeobox transcription factor TLX3. However, the role of TLX3 in T-ALL remains elusive, partly due to the lack of suitable study models. Strikingly, this TLX3-positive subgroup has a high frequency of FLT3 mutations, predominantly FLT3-ITD, in pediatric cases. To investigate this, we generated ex vivo cultured pro-T cells driven by the co-expression of TLX3 and FLT3-ITD, which conferred IL7 independent growth. This model allowed us to confirm that TLX3 expression and FLT3 signaling cooperate to transform T-cells and induce an oncogenic context. Data from this cell model, combined with gene expression data from TLX3 positive T-ALL cases, revealed a strong downregulation of the transcriptional repressor TLE4. Furthermore, TLE4 showed to have a repressive effect on ex vivo TLX3 T-ALL cell growth, likely caused by a partial reversal of the TLX3 transcriptional profile. In conclusion, we developed a TLX3+FLT3-ITD pro-T cell model and used it to illustrate that TLX3 directly represses TLE4 expression, which is beneficial for the oncogenic function of TLX3.

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Fig. 1: FLT3-ITD and TLX3 cooperative ex vivo model represents early stage TLX3 positive phenotype.
Fig. 2: Downregulation of the co-repressor TLE4 is favorable for TLX3 oncogenic activity.
Fig. 3: TLX3 oncogenic function is repressed by TLE4 through its interaction with the EH1 domain of TLX3.
Fig. 4: TLE4 overexpression partially inverts transcriptomic changes induced by TLX3.

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Data availability

Raw RNA-seq and ChIP-seq data performed during this study can be obtained from GSE241640.

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Acknowledgements

We thank Dr Thomas Müller from the Max-Delbrück-Centrum of Molecular Medicine (MDC) in Berlin for kindly providing us with the custom made TLX3 antibody. We thank the VIB flow cytometry core and KU Leuven genomics core for the support in processing the data obtained during this study. The graphical abstract was created using BioRender.com. This work was supported by a FWO fellowship to QVT, a postdoctoral fellowship to SD by the Foundation against Cancer, a postdoctoral FWO fellowship to AV, and a research grant by FWO to JC.

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Author notes

  1. These authors contributed equally: Lukas Lauwereins, Quentin Van Thillo.

Authors and Affiliations

  1. Department of Human Genetics, KU Leuven, Leuven, Belgium

    Lukas Lauwereins, Quentin Van Thillo, Sofie Demeyer, Nicole Mentens, Sarah Provost, Kris Jacobs, Olga Gielen, Lien Boogaerts, Charles E. de Bock, Jan Cools & Alexandra Veloso

  2. Center for Cancer Biology, VIB, Leuven, Belgium

    Lukas Lauwereins, Quentin Van Thillo, Sofie Demeyer, Nicole Mentens, Sarah Provost, Kris Jacobs, Olga Gielen, Lien Boogaerts, Charles E. de Bock, Jan Cools & Alexandra Veloso

  3. Children’s Cancer Institute, UNSW Sydney, Sydney, NSW, Australia

    Charles E. de Bock

  4. Institute Necker Enfants-Malades, INSERM U1151, Paris, France

    Guillaume Andrieu & Vahid Asnafi

  5. Laboratoire d’Onco-Hématologie, Hôpital Necker-Enfants Malades, AP-HP, Paris, France

    Vahid Asnafi

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Contributions

LL and QVT designed and performed experiments, analyzed and interpreted data; NM, SP, OG, LB, and KJ performed experiments; SD performed bioinformatics analyses; AV and JC jointly coordinated the study; CDB involved in scientific discussions and review of the manuscript; GA and VA provided T-ALL adult patient data from GRAALL03/05 and were involved in scientific discussions; LL, QVT, and AV wrote the manuscript. All authors critically read and contributed to the final version of the manuscript.

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Correspondence to Jan Cools or Alexandra Veloso.

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Competing interests

This work was supported by a FWO fellowship to QVT, a postdoctoral fellowship to SD by the Foundation against Cancer, a postdoctoral fellowship to AV by the FWO, and a research grant by FWO to JC.

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Lauwereins, L., Van Thillo, Q., Demeyer, S. et al. TLE4 is a repressor of the oncogenic activity of TLX3 in T-cell acute lymphoblastic leukemia. Leukemia 39, 568–576 (2025). https://doi.org/10.1038/s41375-025-02513-w

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