Efficient and Robust NK-Cell Transduction With Baboon Envelope Pseudotyped Lentivector

oleh: Aurelien B. L. Colamartino, Aurelien B. L. Colamartino, William Lemieux, William Lemieux, Panojot Bifsha, Simon Nicoletti, Simon Nicoletti, Nitin Chakravarti, Joaquín Sanz, Joaquín Sanz, Hugo Roméro, Silvia Selleri, Silvia Selleri, Kathie Béland, Mélanie Guiot, Mélanie Guiot, Camille Tremblay-Laganière, Camille Tremblay-Laganière, Renée Dicaire, Luis Barreiro, Luis Barreiro, Dean A. Lee, Els Verhoeyen, Els Verhoeyen, Elie Haddad, Elie Haddad, Elie Haddad

Format: Article
Diterbitkan: Frontiers Media S.A. 2019-12-01

Deskripsi

NK-cell resistance to transduction is a major technical hurdle for developing NK-cell immunotherapy. By using Baboon envelope pseudotyped lentiviral vectors (BaEV-LVs) encoding eGFP, we obtained a transduction rate of 23.0 ± 6.6% (mean ± SD) in freshly-isolated human NK-cells (FI-NK) and 83.4 ± 10.1% (mean ± SD) in NK-cells obtained from the NK-cell Activation and Expansion System (NKAES), with a sustained transgene expression for at least 21 days. BaEV-LVs outperformed Vesicular Stomatitis Virus type-G (VSV-G)-, RD114- and Measles Virus (MV)- pseudotyped LVs (p < 0.0001). mRNA expression of both BaEV receptors, ASCT1 and ASCT2, was detected in FI-NK and NKAES, with higher expression in NKAES. Transduction with BaEV-LVs encoding for CAR-CD22 resulted in robust CAR-expression on 38.3 ± 23.8% (mean ± SD) of NKAES cells, leading to specific killing of NK-resistant pre-B-ALL-RS4;11 cell line. Using a larger vector encoding a dual CD19/CD22-CAR, we were able to transduce and re-expand dual-CAR-expressing NKAES, even with lower viral titer. These dual-CAR-NK efficiently killed both CD19KO- and CD22KO-RS4;11 cells. Our results suggest that BaEV-LVs may efficiently enable NK-cell biological studies and translation of NK-cell-based immunotherapy to the clinic.