Human cytomegalovirus (HCMV) is a widespread pathogen that usually causes mild or asymptomatic infections in healthy individuals. In contrast, in immunocompromised patients - especially those undergoing high-risk allogeneic stem cell transplantation (SCT) - either a new infection or reactivation of latent HCMV can result in severe, sometimes life-threatening disease. Although several strategies for prophylaxis and treatment have been developed, their clinical use remains limited. Antiviral drugs often cause bone-marrow toxicity, no vaccine has yet been approved, and current virus-specific T-cell therapies require HLA matching between donor and recipient. Together with the virus’s ability to persist lifelong in the host, these factors highlight the need for novel therapeutic approaches.
HCMV-specific chimeric antigen receptor (CAR) T cells provide a promising strategy to overcome these limitations. This project investigates the potential of CMV-specific CAR-T cells to control viral infection in a murine cytomegalovirus (MCMV) model. In particular, we aim to establish messenger RNA (mRNA)-based CAR delivery as a fast, flexible, and safe alternative to conventional viral transduction.
We will synthesize CAR-encoding mRNAs, optimize their electroporation into murine T cells, assess CAR surface expression, and evaluate cytotoxic activity using both reporter cell lines and MCMV-infected target cells. Finally, mRNA-mediated CAR expression will be compared with retroviral delivery regarding efficiency, stability, and functional potency.
We hypothesize that mRNA-engineered CAR-T cells can effectively restrict MCMV infection in vitro while avoiding the risks and time constraints associated with viral gene transfer. If successful, this platform could be adapted to generate HCMV-specific CAR-T cells for use in humans - providing a rapid, HLA-independent, and patient-tailored immunotherapy to prevent CMV complications following allogeneic SCT.
