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

Roellecke K, Virts EL, Einholz R, Edson KZ, Altvater B, Rossig C, von Laer D, Scheckenbach K, Wagenmann M, Reinhardt D, Kramm CM, Rettie AE, Wiek C, Hanenberg H

Forschungsartikel (Zeitschrift) | Peer reviewed

Zusammenfassung

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.

Details zur Publikation

FachzeitschriftGene Therapy (Gene Ther)
Jahrgang / Bandnr. / Volume23
Ausgabe / Heftnr. / Issue7
StatusVeröffentlicht
Veröffentlichungsjahr2016
Sprache, in der die Publikation verfasst istEnglisch
DOI10.1038/gt.2016.38

Autor*innen der Universität Münster

Altvater, Bianca
Klinik für Kinder- und Jugendmedizin - Pädiatrische Hämatologie und Onkologie - (UKM PHO)