Radiocurability by targeting tumor necrosis factor-alpha using a bispecific antibody in carcinoembryonic antigen transgenic mice.
OBJECTIVE
Tumor necrosis factor-alpha (TNF-alpha) 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-alpha to target this cytokine in a CEA-expressing colon carcinoma. We report the evaluation of this strategy in immunocompetent CEA-transgenic mice.


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


RESULTS
In vitro, combined RT plus TNF-alpha 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-alpha, RT plus TNF-alpha, RT alone, and control groups, respectively. This difference was statistically significant when TNF-alpha was targeted with the BsAb (p = 0.03). The addition of exogenous TNF-alpha to RT significantly increased the endogenous TNF-alpha mRNA level, particularly when TNF-alpha was targeted with BsAb (p < 0.01). The percentages of necrotic area were significantly augmented in the RT plus BsAb plus TNF-alpha group.


CONCLUSIONS
These results suggest that targeting TNF-alpha 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.
Larbouret
Christel
C
INSERM, Centre de Recherche en Cancérologie de Montpellier, Université de Montpellier, CRLC Val d'Aurelle-Paul Lamarque, Montpellier, France.
Robert
Bruno
B
Linard
Christine
C
Teulon
Isabelle
I
Gourgou
Sophie
S
Bibeau
Frederic
F
Martineau
Pierre
P
Santoro
Lore
L
Pouget
Jean-Pierre
JP
Pelegrin
Andre
A
Azria
David
D
D000818:Animals; D018033:Antibodies, Bispecific / Q000627:therapeutic use*; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D002470:Cell Survival; D003110:Colonic Neoplasms / Q000276:immunology / Q000532:radiotherapy*; D003131:Combined Modality Therapy / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006801:Humans; D016867:Immunocompromised Host; D051379:Mice; D008822:Mice, Transgenic; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D011897:Random Allocation; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism / Q000627:therapeutic use*; D014410:Tumor Stem Cell Assay / Q000379:methods
D018033:Antibodies, Bispecific; D002272:Carcinoembryonic Antigen; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha