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Chimeric Antigen Receptor-modified T cells targeting EphA2 for the immunotherapy of paediatric bone tumours

Cancer Gene Therapy volume 28, pages 321–334 (2021)Cite this article

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Abstract

Chimeric Antigen Receptor (CAR) T-cell therapy, as an approved treatment option for patients with B cell malignancies, demonstrates that genetic modification of autologous immune cells is an effective anti-cancer regimen. Erythropoietin-producing Hepatocellular receptor tyrosine kinase class A2 (EphA2) is a tumour associated antigen expressed on a range of sarcomas, including paediatric osteosarcoma (OS) and Ewing sarcoma (ES). We tested human EphA2 directed CAR T cells for their capacity to target and kill human OS and ES tumour cells using in vitro and in vivo assays, demonstrating that EphA2 CAR T cells have potent anti-tumour efficacy in vitro and can eliminate established OS and ES tumours in vivo in a dose and delivery route dependent manner. Next, in an aggressive metastatic OS model we demonstrated that systemically infused EphA2 CAR T cells can traffic to and eradicate tumour deposits in murine livers and lungs. These results support further pre-clinical evaluation of EphA2 CAR T cells to inform the design of early phase clinical trial protocols to test the feasibility and safety of this immune cell therapy in paediatric bone sarcoma patients.

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Fig. 1: Expression of tumour-associated antigen target EphA2.
Fig. 2: Vector schematics and LV CAR T-cell transduction efficiencies.
Fig. 3: In vitro assessment of anti-tumour activity of CAR T cells targeted against EphA2.
Fig. 4: In vitro production of IL-2 and IFN-γ by EphA2-targeted CAR T cells.
Fig. 5: Subcutaneous tumour growth and survival curves for OS and ES tumour-bearing mice.
Fig. 6: EphA2 CAR T cells reduce metastatic osteosarcoma tumour burden in mice.
Fig. 7: CAR T cell infiltration into lungs and livers of NSG mice.

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Acknowledgements

The authors gratefully acknowledge the provision of tumour TMA’s from the Sydney Children’s Tumour Bank Network (SCHNTBN).

Funding

This work was funded by project grants received from The Kid’s Cancer Project (TKCP), Cancer Institute NSW through The Kid’s Cancer Alliance (KCA) and The Australian and New Zealand Sarcoma Association (ANZSA).

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Authors and Affiliations

  1. Children’s Cancer Research Unit, Kid’s Research, The Children’s Hospital at Westmead, Westmead, NSW, 2145, Australia

    Kenneth Hsu, Shiloh Middlemiss, Federica Saletta & Belinda Kramer

  2. Department of Bone Marrow Transplant and Cellular Therapy, St. Jude Children’s Research Hospital, Memphis, TN, USA

    Stephen Gottschalk

  3. Children’s Cancer Centre, The Children’s Hospital at Westmead, Westmead, NSW, 2145, Australia

    Geoffrey B. McCowage

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Hsu, K., Middlemiss, S., Saletta, F. et al. Chimeric Antigen Receptor-modified T cells targeting EphA2 for the immunotherapy of paediatric bone tumours. Cancer Gene Ther 28, 321–334 (2021). https://doi.org/10.1038/s41417-020-00221-4

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