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

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 a...

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Bibliographic Details
Published in:	Gene therapy Vol. 23; no. 7; pp. 615 - 626
Main Authors:	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
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 01-07-2016 Nature Publishing Group
Subjects:	42 631/1647/2300/1514 631/250/251 631/45/607/1168 631/61/51/1844 631/80/82/23 692/699/67/1990 Antigens Antigens, CD34 - genetics Antigens, CD34 - metabolism Apoptosis Aryl Hydrocarbon Hydroxylases - genetics Aryl Hydrocarbon Hydroxylases - metabolism Autografts Biomedical and Life Sciences Biomedicine CD19 antigen CD34 antigen Cell Biology Cell surface Cell therapy Cells, Cultured Cellular biology Chimeric antigen receptors Cytochrome P-450 Cytochrome P450 Divergence Enzymes Gene Expression Gene Therapy Genes Genes, Transgenic, Suicide Genetic aspects Genetic engineering Genetic Therapy - methods Genetic vectors Genetic Vectors - genetics Health aspects HEK293 Cells Human Genetics Humans Immunotherapy Immunotherapy, Adoptive - methods Innovations Jurkat Cells Lentivirus - genetics Lymphocytes Lymphocytes T Methods Mutants Mutation Nanotechnology original-article Prodrugs Properties Receptors, Antigen, T-Cell - genetics Receptors, Antigen, T-Cell - metabolism Sarcoma Stem cell transplantation Studies Suicide Suicide genes T cells Terpenes - therapeutic use Terpenoids Testing Toxicity Germany
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0969-7128 1476-5462
DOI:	10.1038/gt.2016.38