A lentiviral vector for the production of T cells with an inducible transgene and a constitutively expressed tumour-targeting receptor

• Published in: Nature biomedical engineering Vol. 7; no. 9; pp. 1063 - 1080
• Main Authors: Reichenbach, Patrick, Giordano Attianese, Greta Maria Paola, Ouchen, Khaoula, Cribioli, Elisabetta, Triboulet, Melanie, Ash, Sarah, Saillard, Margaux, Vuillefroy de Silly, Romain, Coukos, George, Irving, Melita
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-09-2023; Nature Publishing Group
• Subjects: 13/106; 13/31; 13/44; 38/44; 38/5; 42/109; 42/35; 42/41; 59/5; 631/61; 631/61/201; 64/60; 82/80; Antigens; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Interleukin 2; Kinases; Lymphocytes; Lymphocytes T; miRNA; Receptors; Ribonucleic acid; RNA; Transgenes; Tumor microenvironment; Tumors
• ISSN: 2157-846X; 2157-846X
• DOI: 10.1038/s41551-023-01013-5

Vectors that facilitate the engineering of T cells that can better harness endogenous immunity and overcome suppressive barriers in the tumour microenvironment would help improve the safety and efficacy of T-cell therapies for more patients. Here we report the design, production and applicability, in T-cell engineering, of a lentiviral vector leveraging an antisense configuration and comprising a promoter driving the constitutive expression of a tumour-directed receptor and a second promoter enabling the efficient activation-inducible expression of a genetic payload. The vector allows for the delivery of a variety of genes to human T cells, as we show for interleukin-2 and a microRNA-based short hairpin RNA for the knockdown of the gene coding for haematopoietic progenitor kinase 1, a negative regulator of T-cell-receptor signalling. We also show that a gene encoded under an activation-inducible promoter is specifically expressed by tumour-redirected T cells on encountering a target antigen in the tumour microenvironment. The single two-gene-encoding vector can be produced at high titres under an optimized protocol adaptable to good manufacturing practices.
A lentiviral vector incorporating two functionally independent promoters allows for the generation of T cells that express a tumour-directed receptor constitutively and an inducible gene that is expressed only in the presence of a target antigen.