Universal allogeneic CAR T cells engineered with Sleeping Beauty transposons and CRISPR-CAS9 for cancer immunotherapy
| Autor: | Jaitip Tipanee, Ermira Samara-Kuko, Thierry Gevaert, Marinee K. Chuah, Thierry VandenDriessche |
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| Přispěvatelé: | Basic (bio-) Medical Sciences, Division of Gene Therapy & Regenerative Medicine, Faculty of Medicine and Pharmacy, Radiation Therapy, Clinical sciences |
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
Pharmacology
Receptors Chimeric Antigen chimeric antigen receptor transposon T-Lymphocytes Antigens CD19 Hematopoietic Stem Cell Transplantation Receptors Antigen T-Cell leukemia Graft vs Host Disease Immunotherapy Adoptive Biochemistry Genetics and Molecular Biology (miscellaneous) Mice Neoplasms Drug Discovery oncology Genetics Sleeping Beauty Molecular Medicine Animals Immunotherapy CRISPR-Cas Systems CRISPR-Cas9 T cell receptor Molecular Biology |
| Popis: | Allogeneic CD19-specific chimeric antigen receptor (CAR) T cells with inactivated donor T cell receptor (TCR) expression can be used as an "off-the-shelf" therapeutic modality for lymphoid malignancies, thus offering an attractive alternative to autologous, patient-derived T cells. Current approaches for T cell engineering mainly rely on the use of viral vectors. Here, we optimized and validated a non-viral genetic modification platform based on Sleeping Beauty (SB) transposons delivered with minicircles to express CD19-28z.CAR and CRISPR-Cas9 ribonucleoparticles to inactivate allogeneic TCRs. Efficient TCR gene disruption was achieved with minimal cytotoxicity and with attainment of robust and stable CD19-28z.CAR expression. The CAR T cells were responsive to CD19+ tumor cells with antitumor activities that induced complete tumor remission in NALM6 tumor-bearing mice while significantly reducing TCR alloreactivity and GvHD development. Single CAR signaling induced the similar T cell signaling signatures in TCR-disrupted CAR T cells and control CAR T cells. In contrast, TCR disruption inhibited T cell signaling/protein phosphorylation compared with the control CAR T cells during dual CAR/TCR signaling. This non-viral SB transposon-CRISPR-Cas9 combination strategy serves as an alternative for generating next-generation CD19-specific CAR T while reducing GvHD risk and easing potential manufacturing constraints intrinsic to viral vectors. ispartof: MOLECULAR THERAPY vol:30 issue:10 pages:3155-3175 ispartof: location:United States status: published |
| Databáze: | OpenAIRE |
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