TRANSLATION OF TACROLIMUS RESISTANT ANTIVIRAL T CELL PRODUCTS: IN DEPTH-EFFICACY EVALUATION AND HUMAN BASED SAFETY ASSESSMENT

Immunocompromised patients such as patients after solid organ transplantation (SOT) are commonly treated with immunosuppressive drugs to circumvent organ rejection. This entails patients being equipped with a strongly diminished endogenous T cell response against pathogens e.g. viruses. Seasonal and...

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Bibliographic Details
Published in:	Cytotherapy (Oxford, England) Vol. 26; no. 6; pp. e23 - e24
Main Authors:	Burkhardt, L., Wiese, N., Ehlen, L., Schlickeiser, S., Mashreghi, M., Römhild, A., Beltran-Mestres, C., Daher, U., Rothe, M., Arndt, J., Hertel, M., Krebs, A.C., Löwa, A., Hocke, A., Reinke, P., Volk, H., Schmück-Henneresse, M., Amini, L.
Format:	Journal Article
Language:	English
Published:	Elsevier Inc 01-06-2024
Subjects:	antiviral T cells CRISPR-Cas9 Transplantation antiviral T cells CRISPR-Cas9 Transplantation
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Summary:	Immunocompromised patients such as patients after solid organ transplantation (SOT) are commonly treated with immunosuppressive drugs to circumvent organ rejection. This entails patients being equipped with a strongly diminished endogenous T cell response against pathogens e.g. viruses. Seasonal and omnipresent pandemic viruses are major health concerns for these patients. In fact, long-term immunosuppression leads to insensitivity to vaccines, higher rates of bacterial co-infections and results in organ rejection and mortality. In this case, common antiviral drugs cannot combat viral complications. To regenerate a long-term and protective T cell response in immunocompromised patients, we have focused on manufacturing tacrolimus resistant CMV, EBV, and IAV specific T cell products from limited amounts of peripheral blood to be co-administered with immunosuppressive treatment. Our next-generation manufacturing protocol represents a GMP compliant process allowing the production of up to 300 million antiviral T cells within 2 weeks using Grex bioreactors. Knock-out of FKBP12, the functional adaptor protein for the immunosuppressant tacrolimus is achieved by a GMP compliant CRISPR-Cas9 ribonucleoprotein based approach with efficacies of up to 95%. The impact of this genetic modification on T cell products is studied by characterizing effector cytokine production and target specific killing as well as in depth characterization by CITEseq, mass spectrometry based proteomics and epigenetic analyses. We recently gained proof-of-concept results with CITE seq showing functionality of gene edited T cell products in presence of immunosuppressive drugs in comparison to unedited controls. This included also verification of a safety switch using an alternative immunosuppressant, cyclosporine A. Since reproducible pre-clinical safety assessment is of importance for the transition into early phase clinical trials, we gained first insights on T cell product characteristics and safety in co-cultures with primary human cells. This marks a further step towards the development of a relevant human model for T cell product evaluation, which offers particular advantages in the complex environment of cell and gene therapies and immunological questions. All in all, we established a GMP compatible manufacturing process and gathered preclinical data to pave the way for clinical translation of tacrolimus resistant antiviral T cell products.
ISSN:	1465-3249 1477-2566
DOI:	10.1016/j.jcyt.2024.04.048