TY - JOUR
T1 - Efficient lentiviral transduction method to gene modify cord blood CD8(+) T cells for cancer therapy applications
AU - Lo Presti, Vania
AU - Cornel, Annelisa M.
AU - Plantinga, Maud
AU - Dunnebach, Ester
AU - Kuball, Jurgen
AU - Boelens, Jaap Jan
AU - Nierkens, Stefan
AU - van Til, Niek P.
N1 - Funding Information: The authors thank the staff of the Flow Core Facility (Center for Translational Immunology, UMC Utrecht [UMCU], the Netherlands), especially Jeroen van Velzen, for the support and help in the sample analysis. The authors also thank the women for donating the CB and the nurses (Wilhelmina’s Children Hospital, UMCU, the Netherlands) for collecting it. The work done for the manuscript was supported by the Dutch Cancer Society (KWF; grant number 10474 ). Publisher Copyright: © 2021
PY - 2021/6/11
Y1 - 2021/6/11
N2 - 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.
AB - 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.
KW - CD8 T cells
KW - T cell therapy
KW - cord blood
KW - lentiviral transduction
KW - off-the-shelf
UR - http://www.scopus.com/inward/record.url?scp=85103728106&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.omtm.2021.03.015
DO - https://doi.org/10.1016/j.omtm.2021.03.015
M3 - Article
C2 - 33898633
SN - 2329-0501
VL - 21
SP - 357
EP - 368
JO - Molecular Therapy - Methods and Clinical Development
JF - Molecular Therapy - Methods and Clinical Development
ER -