Genetically engineered CAR-NK cell-based approaches to treat cervical cancer

Aim In this project, we engineered natural killer (NK) cells to express chimeric antigen receptors (CAR) to enhance the natural anti-cancer effects of NK cells with the overall aim to develop off-the-shelf therapy approaches against cervical cancer.

Material & Methods Flow cytometry was used to characterize cells derived from cervical cancer patient tissue samples and cervical cancer cell lines (CaSki, HeLa, SiHa) for expression of common cervical cancer antigens. SIN-alpharetroviral vectors were used to generate CAR-NK cells that specifically target the respective antigens. Cervical cancer cells were cultured as monolayers (2D) or spheroids (3D) with CAR-NK cells. Anti-cancer activity of CAR-NK cells was assessed via cytotoxicity (e.g., flow cytometry, Fluoroskan and live cell imaging), and cytokine release assays (e.g., ELISA).

Results Mesothelin was identified as a promising target antigen. Cervical cancer cells that highly expressed Mesothelin were efficiently killed by Mesothelin-specific CAR-NK cells in 2D and 3D culture conditions. In contrast, control NK cells did not exhibit such cytotoxic effects, which suggested strong specificity for target antigens. These findings are consistent with live cell imaging results and increased IFN-γ release into the supernatant upon co-culture of cervical cancer cells with anti-Mesothelin CAR-NK cells.

Summary Taken together, we screened cervical cancer cells for expression of common cervical cancer antigens and show that CAR-NK cells engineered with alpharetroviral vectors efficiently target Mesothelin-positive cervical cancer cells in 2D and 3D conditions. Hence, this strategy displays a promising future approach for the treatment of cervical cancer, which can be expanded to other gynecological cancers.

Publication History

Article published online:
11 October 2022