# LMP2A‐Targeting CAR‐T Cells Equipped With Inducible IL‐18 to Address EBV‐Associated Malignancies

## TL;DR

Scientists developed a new type of CAR-T cell targeting EBV-related cancers by focusing on a specific viral protein and boosting immune response with IL-18.

## Contribution

The novel approach combines CAR-T cells targeting LMP2A with inducible IL-18 to enhance anti-tumor activity in EBV-associated malignancies.

## Key findings

- LMP2A-targeting CAR-T cells and IL-18-equipped TRUCKs specifically recognized EBV-infected cells presenting the CLG peptide.
- LMP2A_iIL-18_TRUCKs showed improved anti-tumor response and potential to recruit bystander immune cells.
- LMP2A_iIL-12_TRUCKs unexpectedly exhibited a different phenotype and non-specific reactivity.

## Abstract

Epstein–Barr virus (EBV) infects up to 95% of the world's population and persists in B cells and epithelial cells. Uncontrolled proliferation of EBV‐infected cells can result in the development of EBV‐associated malignancies, for example, post‐transplant lymphoproliferative disorder (PTLD) or nasopharyngeal cancer (NPC). It is estimated that 1.8% of deaths due to cancer worldwide are associated with EBV, and the treatment options are limited. As a new therapeutic approach, we developed chimeric antigen receptor T cells targeting EBV‐derived latent membrane protein 2A (LMP2A_CAR‐Ts). To enable specific elimination of malignant B cells infected with the intracellular pathogen, we utilised T‐cell receptor (TCR)‐like specificity to generate a CAR against the LMP2A‐derived peptide CLGGLLTMV (CLG) presented in the context of HLA‐A*02:01. To increase functionality in the tumour microenvironment, LMP2A_CAR‐Ts were additionally equipped with inducible release of IL‐12 (LMP2A_iIL‐12_TRUCKs) or IL‐18 (LMP2A_iIL‐18_TRUCKs). LMP2A_CAR‐Ts and LMP2A_iIL‐18_TRUCKs specifically recognised HLA‐A*02:01
+ EBV‐transformed B‐lymphoblastoid cell lines and HLA‐A*02:01
+ cells loaded with CLG peptide (A02_CLG+ cells), proving their target specificity, while, unexpectedly, LMP2A_iIL‐12_TRUCKs exhibited a disparate TEM and NK‐like phenotype and A02_CLG‐independent reactivity. In contrast, LMP2A_CAR‐Ts and LMP2A_iIL‐18_TRUCKs effectively induced T‐cell signalling, activation, release of cytotoxic mediators, including IL‐18 by LMP2A_iIL‐18_TRUCKs and mediated cytotoxicity in a target‐specific manner. Our results demonstrate that LMP2A_CAR‐Ts and LMP2A_iIL‐18_TRUCKs specifically recognise the EBV‐derived CLG peptide presented in the context of HLA‐A*02:01. Especially, LMP2A_iIL‐18_TRUCKs with an even improved anti‐tumour response, as well as the potential to recruit bystander immune cells and overcome EBV‐mediated immune evasion strategies, might serve as a novel treatment option for various EBV‐associated malignancies.