The Tol2 transposon system mediates the genetic engineering of T-cells with CD19-specific chimeric antigen receptors for B-cell malignancies Journal Article
| Authors: | Tsukahara, T.; Iwase, N.; Kawakami, K.; Iwasaki, M.; Yamamoto, C.; Ohmine, K.; Uchibori, R.; Teruya, T.; Ido, H.; Saga, Y.; Urabe, M.; Mizukami, H.; Kume, A.; Nakamura, M.; Brentjens, R.; Ozawa, K. |
| Article Title: | The Tol2 transposon system mediates the genetic engineering of T-cells with CD19-specific chimeric antigen receptors for B-cell malignancies |
| Abstract: | Engineered T-cell therapy using a CD19-specific chimeric antigen receptor (CD19-CAR) is a promising strategy for the treatment of advanced B-cell malignancies. Gene transfer of CARs to T-cells has widely relied on retroviral vectors, but transposon-based gene transfer has recently emerged as a suitable nonviral method to mediate stable transgene expression. The advantages of transposon vectors compared with viral vectors include their simplicity and cost-effectiveness. We used the Tol2 transposon system to stably transfer CD19-CAR into human T-cells. Normal human peripheral blood lymphocytes were co-nucleofected with the Tol2 transposon donor plasmid carrying CD19-CAR and the transposase expression plasmid and were selectively propagated on NIH3T3 cells expressing human CD19. Expanded CD3 + T-cells with stable and high-level transgene expression (∼95%) produced interferon-γ upon stimulation with CD19 and specifically lysed Raji cells, a CD19 + human B-cell lymphoma cell line. Adoptive transfer of these T-cells suppressed tumor progression in Raji tumor-bearing Rag2 -/- γc -/- immunodeficient mice compared with control mice. These results demonstrate that the Tol2 transposon system could be used to express CD19-CAR in genetically engineered T-cells for the treatment of refractory B-cell malignancies. |
| Keywords: | controlled study; unclassified drug; human cell; nonhuman; cd8+ t lymphocyte; t lymphocyte; mouse; mus; gene expression; animal experiment; animal model; in vitro study; tumor xenograft; gene transfer; cost effectiveness analysis; b cell lymphoma; genetic engineering; cd4+ t lymphocyte; retrovirus vector; chimeric antigen receptor; immunostimulation; large cell lymphoma; tumor growth; adoptive immunotherapy; plasmid vector; transposon; peripheral lymphocyte; nonviral gene delivery system; nonviral gene therapy; cancer gene therapy; cell based gene therapy; cell engineering; human; priority journal; article; raji cell line; 3t3 cell line; cd19 specific chimeric antigen receptor; tol2 transposon vector; inverse polymerase chain reaction; k562 cell line |
| Journal Title: | Gene Therapy |
| Volume: | 22 |
| Issue: | 2 |
| ISSN: | 0969-7128 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2015-02-01 |
| Start Page: | 209 |
| End Page: | 215 |
| Language: | English |
| DOI: | 10.1038/gt.2014.104 |
| PROVIDER: | scopus |
| PUBMED: | 25427612 |
| PMCID: | PMC5548386 |
| DOI/URL: | |
| Notes: | Export Date: 2 March 2015 -- Source: Scopus |
