Efficient lentiviral transduction method to gene modify cord blood CD8+ T cells for cancer therapy applications

• Published in: Molecular therapy. Methods & clinical development 2021-06, Vol.21, p.357-368
• Main Authors: Lo Presti, Vania, Cornel, Annelisa M., Plantinga, Maud, Dünnebach, Ester, Kuball, Jurgen, Boelens, Jaap Jan, Nierkens, Stefan, van Til, Niek P.
• Format: Article
• Language: English
• Subjects: CD8+ T cells; cord blood; lentiviral transduction; off-the-shelf; Original; T cell therapy
• ISSN: 2329-0501; 2329-0501
• DOI: 10.1016/j.omtm.2021.03.015

Adoptive T cell therapy utilizing tumor-specific autologous T cells has shown promising results for cancer treatment. However, the limited numbers of autologous tumor-associated antigen (TAA)-specific T cells and the functional aberrancies, due to disease progression or treatment, remain factors that may significantly limit the success of the therapy. The use of allogeneic T cells, such as umbilical cord blood (CB) derived, overcomes these issues but requires gene modification to induce a robust and specific anti-tumor effect. CB T cells are readily available in CB banks and show low toxicity, high proliferation rates, and increased anti-leukemic effect upon transfer. However, the combination of anti-tumor gene modification and preservation of advantageous immunological traits of CB T cells represent major challenges for the harmonized production of T cell therapy products. In this manuscript, we optimized a protocol for expansion and lentiviral vector (LV) transduction of CB CD8+ T cells, achieving a transduction efficiency up to 83%. Timing of LV treatment, selection of culture media, and the use of different promoters were optimized in the transduction protocol. LentiBOOST was confirmed as a non-toxic transduction enhancer of CB CD8+ T cells, with minor effects on the proliferation capacity and cell viability of the T cells. Positively, the use of LentiBOOST does not affect the functionality of the cells, in the context of tumor cell recognition. Finally, CB CD8+ T cells were more amenable to LV transduction than peripheral blood (PB) CD8+ T cells and maintained a more naive phenotype. In conclusion, we show an efficient method to genetically modify CB CD8+ T cells using LV, which is especially useful for off-the-shelf adoptive cell therapy products for cancer treatment. [Display omitted] The manuscript proposes an efficient method to gene modify cord blood-derived CD8+ T cells using lentiviral vectors and a transduction enhancer (LentiBOOST). Future application of these findings can improve the generation of allogeneic T cell therapies for the treatment of cancer.