Kinetics of cell labelling and thymidine replacement after continuous infusion of halogenated pyrimidines in vivo

Abstract

Purpose :Iododeoxyuridine (IdUrd) is a halogenated pyrimidine which has been recognized as a clinical radiosensitizer. It is generally agreed that the extent of radiosensitization correlates with the degree of thymidine substitution in DNA. Controversy exists regarding the optimal administration schedule to achieve maximum radiosensitization. To obtain more information relating to this problem, we present experiments on an in vivo human tumor xenograft continuously exposed to a fixed serum concentration of halogenated pyrimidines so as to study the kinetics of cell labeling and thymidine replacement. Methods and Materials : Human colon tumor (HCT-116) cells were injected subcutaneously into nude mice. After 10 days, most animals (> 90%) developed measurable tumor nodules with a volume doubling time of 5 ± 1 days. Once the tumors reached a cross-sectional area of 0.25–0.30 cm 2, miniosmotic pumps were implanted to deliver a dose of 100 mg/kg/day of IdUrd by continuous infusion. After an IdUrd exposure time of 1–7 days, blood and tumor tissue were collected. Results : The steady state serum IdUrd concentration was 0.95 ± 0.1 μM, which is a clinically relevant concentration for a prolonged continuous intravenous infusion. The tumor cell potential doubling time (T pot) was 25 ± 2 h. The percent IdUrd thymidine replacement and the fraction of cells labeled, followed exponential saturation kinetics with a halflife of 33 ± 9 and 27 ± 2 h, respectively. After 5 days of exposure (∼- 5 × T pot, the thymidine replacement in tumor cells was 2.0 ± 0.2% and the fraction of tumor cells labeled was 94 ± 1%. Immunohistochemical staining of IdUrd labeled tumor tissues showed an exposure dependent gradient of cellular labeling that was initially highest in regions close to blood vessels. After 4 days of exposure at 100 mg/kg/day, there was an increase in the fraction of cells in Go + G, and a decrease in the S phase population, suggesting a block between G, and S phase. Conclusion : We conclude that the in vivo kinetics of IdUrd thymidine replacement and fraction of cells labeled after continuous exposure followed exponential saturation kinetics with a halflife of approximately the potential doubling time of the tumor cell population. Simple modeling suggests that some form of prolonged, or briefly interrupted, continuous infusion should be considered for clinical administration because such schedules would leave fewer cells unsensitized than shorter infusions would. Even 10% of unlabeled clonogenic cells could explain the lack of dramatic clinical successes with IdUrd or BrdUrd sensitization.