Radiocurability by Targeting Tumor Necrosis Factor-α Using a Bispecific Antibody in Carcinoembryonic Antigen Transgenic Mice

Purpose

Tumor necrosis factor-α (TNF-α) enhances radiotherapy (RT) killing of tumor cells in vitro and in vivo. To overcome systemic side effects, we used a bispecific antibody (BsAb) directed against carcinoembryonic antigen (CEA) and TNF-α to target this cytokine in a CEA-expressing colon carcinoma. We report the evaluation of this strategy in immunocompetent CEA-transgenic mice.

Methods and Materials

The murine CEA-transfected colon carcinoma MC-38 was used for all experiments. In vitro, clonogenic assays were performed after RT alone, TNF-α alone, and RT plus TNF-α. In vivo, the mice were randomly assigned to treatment groups: control, TNF-α, BsAb, BsAb plus TNF-α, RT, RT plus TNF-α, and RT plus BsAb plus TNF-α. Measurements of endogenous TNF-α mRNA levels and evaluation of necrosis (histologic evaluation) were assessed per treatment group.

Results

In vitro, combined RT plus TNF-α resulted in a significant decrease in the survival fraction at 2 Gy compared with RT alone (p < 0.00001). In vivo, we observed a complete response in 5 (50%) of 10, 2 (20%) of 10, 2 (18.2%) of 11, and 0 (0%) of 12 treated mice in the RT plus BsAb plus TNF-α, RT plus TNF-α, RT alone, and control groups, respectively. This difference was statistically significant when TNF-α was targeted with the BsAb (p = 0.03). The addition of exogenous TNF-α to RT significantly increased the endogenous TNF-α mRNA level, particularly when TNF-α was targeted with BsAb (p < 0.01). The percentages of necrotic area were significantly augmented in the RT plus BsAb plus TNF-α group.

Conclusion

These results suggest that targeting TNF-α with the BsAb provokes RT curability in a CEA-expressing digestive tumor syngenic model and could be considered as a solid rationale for clinical trials.

Bispecific antibody, Tumor necrosis factor-α, Radiotherapy enhancement, CEA-transgenic mice, Synergism