Background: Strategies that enhance the function of T cells are critical for immunotherapy. Targeted delivery of T cells through BiTE (bispecific T-cell engager) platform to cancerous tissues shows potential in sparing unaffected tissues. However, it has been a major challenge for cells penetration in solid tumor tissues due to the complicated tumor microenvironment. Activated T cells expression integrin, which is the target of peptide RGD. Peptide iRGD (CRGDK/RGPD/EC) increased vascular and tissue permeability in a tumor-specific and neuropilin-1-dependent manner, allowing co-administered drugs to penetrate into extravascular tumor tissue. Recombinant protein anti-EGFR-iRGD was purified and examinated.
Methods: Recombinant protein anti-EGFR-iRGD consisting of an anti-EGFR VHH (the variable domain from the heavy chain of the antibody) fused to iRGD, a tumor-specific binding peptide with high permeability were expressed in E. coli BL21 (DE3) and purified by nickel-nitrilotriacetic acid affinity chromatography. We use tumor cell lines and mice to analyze the targeting, penetrating and antitumor activity of antigen-secific T cells together with recombinant protein.
Results: We have successfully constructed a recombinant protein named anti-EGFR-iRGD, a dual targets of EGFR and integrin and high permeable protein. It could spread extensively throughout both the multicellular spheroids and the tumor mass. The recombinant protein anti-EGFR-iRGD could help more T cells infiltrating into tumor mass and also exhibited antitumor activity in tumor cell lines and mice.
Note: This abstract was not presented at the conference.
Conclusions: Our results provide impetus for further studies for potentially using iRGD based fusion protein anti-EGFR-iRGD with immune therapy regimens for enhancing therapy of gastric cancer patients.
Citation Format: Huizi Sha, Baorui Liu. A tumor-penetrating recombinant protein anti-EGFR-iRGD enhance efficacy of antigen-specific CTL in gastric cancer in vivo. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr A20.