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Translational Therapeutics

Ponatinib delays the growth of solid tumours by remodelling immunosuppressive tumour microenvironment through the inhibition of induced PD-L1 expression

British Journal of Cancer volume 129, pages 1007–1021 (2023)Cite this article

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Abstract

Background

Therapeutic modalities including chemo, radiation, immunotherapy, etc. induce PD-L1 expression that facilitates the adaptive immune resistance to evade the antitumour immune response. IFN-γ and hypoxia are some of the crucial inducers of PD-L1 expression in tumour and systemic microenvironment which regulate the expression of PD-L1 via various factors including HIF-1α and MAPK signalling. Hence, inhibition of these factors is crucial to regulate the induced PD-L1 expression and to achieve a durable therapeutic outcome by averting the immunosuppression.

Methods

B16-F10 melanoma, 4T1 breast carcinoma, and GL261 glioblastoma murine models were established to investigate the in vivo antitumour efficacy of Ponatinib. Western blot, immunohistochemistry, and ELISA were performed to determine the effect of Ponatinib on the immunomodulation of tumour microenvironment (TME). CTL assay and flow cytometry were such as p-MAPK, p-JNK, p-Erk, and cleaved caspase-3 carried out to evaluate the systemic immunity induced by Ponatinib. RNA sequencing, immunofluorescence and Western blot analysis were used to determine the mechanism of PD-L1 regulation by Ponatinib. Antitumour immunity induced by Ponatinib were compared with Dasatinib.

Results

Here, Ponatinib treatment delayed the growth of tumours by inhibiting PD-L1 and modulating TME. It also downregulated the level of PD-L1 downstream signalling molecules. Ponatinib enhanced the CD8 T cell infiltration, regulated Th1/Th2 ratio and depleted tumour associated macrophages (TAMs) in TME. It induced a favourable systemic antitumour immunity by enhancing CD8 T cell population, tumour specific CTL activity, balancing the Th1/Th2 ratio and lowering PD-L1 expression. Ponatinib inhibited FoxP3 expression in tumour and spleen. RNA sequencing data revealed that Ponatinib treatment downregulated the genes related to transcription including HIF-1α. Further mechanistic studies showed that it inhibited the IFN-γ and hypoxia induced PD-L1 expression via regulating HIF-1α. Dasatinib was used as control to prove that Ponatinib induced antitumour immunity is via PD-L1 inhibition mediated T cell activation.

Conclusions

RNA sequencing data along with rigorous in vitro and in vivo studies revealed a novel molecular mechanism by which Ponatinib can inhibit the induced PD-L1 levels via regulating HIF-1α expression which leads to modulation of tumour microenvironment. Thus, our study provides a novel therapeutic insight of Ponatinib for the treatment of solid tumours where it can be used alone or in combination with other drugs which are known to induce PD-L1 expression and generate adaptive resistance.

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Fig. 1: Tumour growth inhibition study in B16-F10 melanoma and 4T1 breast carcinoma model.
Fig. 2: CD8 T cell infiltration and PD-L1 inhibition.
Fig. 3: Modulation of TME by Ponatinib treatment.
Fig. 4: Ponatinib enhanced T cell populations and CTL activity.
Fig. 5: Modulation of splenic microenvironment.
Fig. 6: Ponatinib inhibits the PD-L1 expression via HIF-1α downregulation.
Fig. 7: Modulation of TME by Ponatinib in comparison with Dasatinib.

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Data availability

All the materials used to produce the data in this study are available from the corresponding authors upon reasonable request.

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Acknowledgements

We would like to acknowledge the animal facility at National Institute of Immunology, Delhi for allowing to perform the animal studies. AB acknowledges IIT Delhi for providing her doctoral fellowship. We also acknowledge Mr. Amit Kumar from ILBS for helping in IHC slide preparation.

Funding

This research was supported through a project by Department of Biotechnology sanctioned to JB (Grant No. BT/PR29866/NNT/28/1586/2018) and a faculty interdisciplinary research project by Indian Institute of Technology Delhi (Grant No. MI02200G) (to JB).

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Authors and Affiliations

  1. Centre for Biomedical Engineering, Indian Institute of Technology Delhi, Delhi, India

    Anjali Barnwal & Jayanta Bhattacharyya

  2. Department of Biomedical Engineering, All India Institute of Medical Science, Delhi, India

    Anjali Barnwal & Jayanta Bhattacharyya

  3. National Institute of Immunology, Delhi, India

    Rohini Tamang &  Sanjeev Das

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AB and JB conceived and designed the experiments. AB performed the experiments. AB and JB analysed the data and wrote the paper. SD and RT helped with the animal studies. All authors discussed the results and commented on the manuscript.

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Correspondence to Jayanta Bhattacharyya.

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Barnwal, A., Tamang, R., Sanjeev Das et al. Ponatinib delays the growth of solid tumours by remodelling immunosuppressive tumour microenvironment through the inhibition of induced PD-L1 expression. Br J Cancer 129, 1007–1021 (2023). https://doi.org/10.1038/s41416-023-02316-9

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