LMP2A-Targeting CAR-T Cells Equipped With Inducible IL-18 to Address EBV-Associated Malignancies.

Area of Science:

• Immunology
• Oncology
• Virology

Background:

• Epstein-Barr virus (EBV) infects a majority of the global population, persisting in B cells and epithelial cells.
• Uncontrolled proliferation of EBV-infected cells leads to malignancies like post-transplant lymphoproliferative disorder (PTLD) and nasopharyngeal cancer (NPC).
• EBV is linked to 1.8% of global cancer deaths, with limited treatment options available.

Purpose of the Study:

• To develop a novel therapeutic approach for EBV-associated malignancies using chimeric antigen receptor T cells (CAR-Ts).
• To engineer CAR-Ts with T-cell receptor (TCR)-like specificity against EBV-derived latent membrane protein 2A (LMP2A) peptide CLGGLLTMV (CLG) presented by HLA-A*02:01.
• To enhance CAR-T functionality in the tumor microenvironment by incorporating inducible IL-12 or IL-18 release.

Main Methods:

• Development of LMP2A-specific CAR-Ts targeting the CLG peptide presented in the context of HLA-A*02:01.
• Engineering of LMP2A_CAR-Ts with inducible IL-12 (LMP2A_iIL-12_TRUCKs) or IL-18 (LMP2A_iIL-18_TRUCKs).
• Assessment of target specificity, T-cell signaling, activation, cytokine release, and cytotoxicity against EBV-transformed B-lymphoblastoid cell lines and peptide-loaded cells.

Main Results:

• LMP2A_CAR-Ts and LMP2A_iIL-18_TRUCKs demonstrated specific recognition of HLA-A*02:01+ EBV-transformed B-lymphoblastoid cell lines and A02_CLG+ cells.
• LMP2A_CAR-Ts and LMP2A_iIL-18_TRUCKs induced T-cell signaling, activation, and cytotoxic mediator release, including IL-18 from LMP2A_iIL-18_TRUCKs.
• LMP2A_iIL-12_TRUCKs exhibited unexpected T_EM and NK-like phenotypes with A02_CLG-independent reactivity, while LMP2A_CAR-Ts and LMP2A_iIL-18_TRUCKs mediated target-specific cytotoxicity.

Conclusions:

• LMP2A_CAR-Ts and LMP2A_iIL-18_TRUCKs effectively recognize the EBV-derived CLG peptide in the context of HLA-A*02:01.
• LMP2A_iIL-18_TRUCKs show enhanced anti-tumor response, potential to recruit immune cells, and overcome EBV immune evasion.
• These engineered CAR-T cells represent a promising novel therapeutic strategy for various EBV-associated malignancies.