DETERMINATION OF THE RADIOBIOLOGICALLY HYPOXIC FRACTION IN MULTICELLULAR SPHEROIDS FROM DATA ON THE UPTAKE OF [H-3] FLUOROMISONIDAZOLE

Fluoromisonidazole [1-(2-nitroimidazolyl)-2-hydroxy-3-fluoro-propane, FMISO] shows promise as a hypoxia imaging agent: it binds preferentially to anoxic cells in monolayers in vitro and accumulates in radiobiologically hypoxic tumors in vivo. The multicellular spheroid model was used to determine if the radiobiologically hypoxic fraction could be predicted from data on the uptake of FMISO. Chinese hamster V79-171b spheroids approximately 500 mu m in diameter were exposed to 50 mM [H-3]FMISO for 1 to 6 h under aerobic (5% CO2 in air), hypoxic (5% CO2, 5% O-2, in N-2) or anoxic (5% CO2 in N-2) conditions and FMISO uptake was measured. Uptake in anoxic spheroids was similar to that in anoxic cell monolayers, while there was virtually no uptake in aerobic spheroids. A mathematical model was developed to calculate the radiobiologically hypoxic fraction in the hypoxic spheroids from the data on FMISO uptake. A radiobiologically hypoxic fraction of 15% was obtained, consistent with that determined from radiation survival assays (17%) and measurements of oxygen consumption (22%). We conclude that the rate of FMISO uptake in V79-171b spheroids correlates with the radiobiologically hypoxic fraction. Furthermore, the radiobiologically hypoxic fraction can be calculated from data on FMISO uptake if the dependence of FMISO uptake on oxygen concentration is known for a given tumor cell type.