Optimized human CYP4B1 in combination with the alkylator prodrug 4-ipomeanol serves as a novel suicide gene system for adoptive T-cell therapies

K. Roellecke, E. L. Virts, R. Einholz, K. Z. Edson, B. Altvater, C. Rossig, D. Von Laer, K. Scheckenbach, M. Wagenmann, D. Reinhardt, C. M. Kramm, A. E. Rettie, C. Wiek, H. Hanenberg

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Engineering autologous or allogeneic T cells to express a suicide gene can control potential toxicity in adoptive T-cell therapies. We recently reported the development of a novel human suicide gene system that is based on an orphan human cytochrome P450 enzyme, CYP4B1, and the naturally occurring alkylator prodrug 4-ipomeanol. The goal of this study was to systematically develop a clinically applicable self-inactivating lentiviral vector for efficient co-expression of CYP4B1 as an ER-located protein with two distinct types of cell surface proteins, either MACS selection genes for donor lymphocyte infusions after allogeneic stem cell transplantation or chimeric antigen receptors for retargeting primary T cells. The U3 region of the myeloproliferative sarcoma virus in combination with the T2A site was found to drive high-level expression of our CYP4B1 mutant with truncated CD34 or CD271 as MACS suitable selection markers. This lentiviral vector backbone was also well suited for co-expression of CYP4B1 with a codon-optimized CD19 chimeric antigen receptor (CAR) construct. Finally, 4-ipomeanol efficiently induced apoptosis in primary T cells that co-express mutant CYP4B1 and the divergently located MACS selection and CAR genes. In conclusion, we here developed a clinically suited lentiviral vector that supports high-level co-expression of cell surface proteins with a potent novel human suicide gene.

Original languageEnglish (US)
Pages (from-to)615-626
Number of pages12
JournalGene Therapy
Volume23
Issue number7
DOIs
StatePublished - Jul 1 2016
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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    Roellecke, K., Virts, E. L., Einholz, R., Edson, K. Z., Altvater, B., Rossig, C., Von Laer, D., Scheckenbach, K., Wagenmann, M., Reinhardt, D., Kramm, C. M., Rettie, A. E., Wiek, C., & Hanenberg, H. (2016). Optimized human CYP4B1 in combination with the alkylator prodrug 4-ipomeanol serves as a novel suicide gene system for adoptive T-cell therapies. Gene Therapy, 23(7), 615-626. https://doi.org/10.1038/gt.2016.38