PET with the Zr-89-Labeled Transforming Growth Factor-beta Antibody Fresolimumab in Tumor Models

Transforming growth factor-beta (TGF-beta) promotes cancer invasion and metastasis and is therefore a potential drug target for cancer treatment. Fresolimumab, which neutralizes all mammalian active isoforms of TGF-beta, was radiolabeled with Zr-89 for PET to analyze TGF-beta expression, antibody tumor uptake, and organ distribution. Methods: Zr-89 was conjugated to fresolimumab using the chelator N-succinyldesferrioxamine-B-tetrafluorphenol. Zr-89-fresolimumab was analyzed for conjugation ratio, aggregation, radiochemical purity, stability, and immunoreactivity. Zr-89-fresolimumab tumor uptake and organ distribution were assessed using 3 protein doses (10, 50, and 100 mu g) and compared with In-111-IgG in a human TGF-beta-transfected Chinese hamster ovary xenograft model, human breast cancer MDA-MB-231 xenograft, and metastatic model. Latent and active TGF-beta 1 expression was analyzed in tissue homogenates with enzyme-linked immunosorbent assay. Results: Zr-89 was labeled to fresolimumab with high specific activity (>1 GBq/mg), high yield, and high purity. In vitro validation of 89Zr-fresolimumab showed a fully preserved immunoreactivity and long (>1 wk) stability in solution and in human serum. In vivo validation showed an Zr-89-fresolimumab distribution similar to IgG in most organs, except for a higher uptake in the liver in all mice and higher kidney uptake in the 10-mu g group. Zr-89-fresolimumab induced no toxicity in mice; it accumulated in primary tumors and metastases in a manner similar to IgG. Both latent and active TGF-beta was detected in tumor homogenates, whereas only latent TGF-beta could be detected in liver homogenates. Remarkably high Zr-89-fresolimumab uptake was seen in sites of tumor ulceration and in scar tissue, processes in which TGF-beta is known to be highly active. Conclusion: Fresolimumab tumor uptake and organ distribution can be visualized and quantified with Zr-89-fresolimumab PET. This technique will be used to guide further clinical development of fresolimumab and could possibly identify patients most likely to benefit.