Employment of NKG2D as chimeric receptor of immune cells and chemokines CXCL9/10 for enhancement of therapeutic effect on colorectal cancer cells

Employment of NKG2D as chimeric receptor of immune cells and chemokines CXCL9/10 for enhancement of therapeutic effect on colorectal cancer cells

Project goal: Objective 1: To evaluate the cytotoxic potential of immune cells expressing the chimeric NKG2D receptor against colorectal cancer (CRC) cells.

Objective 2: To perform a comparative analysis of the contribution of different chemokines to the enhancement of CAR-NKG2D-T cell infiltration efficiency using various colorectal cancer cell models.

Project description: This project focuses on the development of a novel CAR-T cell-based strategy for colorectal cancer (CRC) by combining NKG2D-directed cytotoxicity with enhanced tumor infiltration mediated by chemokines CXCL9 and CXCL10. The approach targets immunologically “cold” CRC subtypes that are poorly responsive to standard immunotherapies, aiming to improve immune cell recruitment and tumor eradication.

Project facilitators: PI: Nikolai Barlev, Co-PI: Dinara Begimbetova. Aleksei Petukhov, Shynggys Tursymbek, Alina Ershova, Aliya Kabasheva

Realisation period: 2024-2026

Expected results: The project is expected to generate CAR-NKG2D-T cells with enhanced cytotoxicity and tumor infiltration capacity. It will also identify the most effective chemokines for improving CAR-T cell migration into CRC models and clarify the influence of p53 status on ligand expression.

Methodology: The study employs genetic engineering of T cells using in vitro–transcribed mRNA, followed by co-culture with CRC models. Cytotoxicity will be assessed using RTCA assays, while gene expression of NKG2D ligands and chemokines will be quantified by qPCR and flow cytometry under genotoxic stress. Cell migration will be evaluated using soft agar–based assays.

Contacts: Nikolai Barlev, nikolai.barlev@nu.edu.kz