88P - Evaluating the infiltration of anti-NKG2DL CAR-T cells into a 3D cell culture developed in a Vitvo cartridge bioreactor

Background

Oncological diseases take the second place in the structure of total mortality. Traditionally, the choice of a method of treating oncological diseases is determined by nosology. In this regard, methods that combine an individual approach and versatility attract much attention. Adoptive cell therapy, including CAR-T therapy, is such a tool. anti-NKG2DL CAR-T cells and NK cells are currently being evaluated for safety and efficacy for targeting tumors positive for NKG2D ligands. The aim of the work was to study the effectiveness of anti-NKG2DL CAR-T cells on PC3M tumor cells.

Methods

Healthy donor peripheral blood mononuclear cells (PBMCs) were used to generate anti-NKG2DL CAR-T cells using lentivirus transduction, the transduction efficiency and percentage of resulting CAR-T cells were evaluated by flow cytometry on a FACS Aria III instrument using anti-NKG2D PE antibodies. As a model of solid tumors 3D Vitvo cartridge bioreactor (Rigenerand) seeded with PC3M-Katushka2S was used. Penetration of anti-NKG2DL CAR-T cells into the 3D cell model was assessed by confocal microscopy.

Results

Transduction efficiency of T-cells with lentivirus was 26%. It was found that CAR-T cells, after being added to the cartridge bioreactor, migrate to the membrane with PC3M cells and further are also predominantly located on the membrane or near it. At the same time, T cells, on the contrary, were located mainly in suspension. By the end of the incubation period of 120 hours in the experimental sample, the presence of CAR-T cells provided a longer decrease in the number of PC3M cells and a slower recovery of their number compared to the control sample (T cells).

Conclusions

Anti-NKG2DL CAR-T cells are able to migrate to the membrane with PC3M cells and reduce their proliferative activity. The study has been performed according to the Kazan Federal University Strategic Academic Leadership Program (PRIORITY-2030).

Editorial acknowledgement

Clinical trial identification

Legal entity responsible for the study

The authors.

Funding

The study has been funded by RSF grant 22-74-10076 and performed according to the Kazan Federal University Strategic Academic Leadership Program (PRIORITY 2030).

Disclosure

All authors have declared no conflicts of interest.