Marianne Kelley

Modulation of tumor oxygenation and radio sensitivity by a perfluorooctylbromide emulsion

The effect of a concentrated perfluorooctylbromide emulsion (Oxygent) on the radiosensitivity and oxygenation of solid tumors was examined using EMT6 mammary tumors in BALB/c mice and BA1112 rhabdomyosarcomas in WAG/rij rats. Treatment with Oxygent plus carbogen or oxygen breathing increased the radiosensitivity of both tumors. Analysis of tumor cell survival data and polarographic measurements of intratumoral pO2 indicated that this potentiation reflected an increase in the proportion of well-oxygenated tumor cells. Treatments with carbogen breathing alone, with Oxygent plus air-breathing, or with a vehicle emulsion containing all the components except the perfluorocarbon did not produce comparable improvements in tumor radiosensitivity. Concentrated perfluorooctylbromide emulsions appear to warrant further development and preclinical testing as adjuncts to cancer therapy.

RSR13, a synthetic allosteric modifier of hemoglobin, as an adjunct to radiotherapy: Preliminary studies with EMT6 cells and tumors and normal tissues in mice

RSR13, 2[4-[[(3,5dimethylanilino)carbonyl]methyl]phenoxy]-2-methylpropion ic acid, a synthetic allosteric modifier of hemoglobin, reduces the affinity of hemoglobin for oxygen. The experiments reported here examined the effect of treatment with RSR13, combined with oxygen breathing, on the radiation response of EMT6 mammary tumors in BALB/c mice and of two normal tissues. RSR13 plus oxygen breathing increased the response of EMT6 tumors to irradiation. RSR13 had no discernible effects on tumors rendered maximally hypoxic by nitrogen asphyxiation, no discernible cytotoxic effects in EMT6 tumors, and no effect on the viability or radiation response of EMT6 cells in vitro under either aerobic or hypoxic conditions. The effects of RSR13 therefore reflect changes in tumor oxygenation, rather than a direct cytotoxic or radiosensitizing effect of the drug. RSR13 plus oxygen reduced the hypoxic fraction to 9% from the value of 24% found in both air-breathing and oxygen-breathing mice. Treatment with RSR13 plus oxygen did not alter the radiation response of the bone marrow progenitor cells (CFU-S) or acute radiation reactions in the skin. The improvement in tumor radiation response produced by treatment with RSR13 plus oxygen, combined with the absence of enhanced radiation reactions in the normal tissues, support further testing of RSR13 as an adjunct to radiotherapy.

Preclinical studies of a perfluorochemical emulsion as an adjunct to radiotherapy

Tumor growth and tumor cell survival endpoints were used to examine the effects of a perfluorochemical emulsion, Fluosol-DA, 20%, and carbogen (95% O2/5% CO2) on EMT6 mouse mammary tumors in BALB/c mice. These studies defined the effects of the Fluosol dose on the hematocrit and fluorocrit of the mice and on the radiation response of the tumors. The effect of varying the duration of carbogen breathing before irradiation was examined; times of 5-60 min gave similar enhancements of tumor radiosensitivity. Potentiating effects were not observed when the tumors were irradiated 1-3 days after Fluosol injection, probably reflecting the observed clearance of the perfluorochemicals from the circulating blood. Fluosol injected 30 min-2 days before irradiation did not alter the radiation response of tumors in air-breathing or N2-asphyxiated mice. Together, these studies provided additional support for the hypothesis that the potentiation of tumor radiation response observed after treatment with Fluosol plus carbogen results from changes in O2 delivery to the hypoxic tumor cells by oxygenated perfluorochemical particles. This confirms the conclusion drawn on the basis of the observed changes in the tumor cell survival curve. Studies of tumor cell survival, tumor cell yield, tumor growth, and artificial lung metastasis formation revealed no effects of Fluosol treatment (without irradiation) on tumor progression or metastasis. Studies examining the effects of Fluosol plus carbogen on the growth of tumors irradiated with 5 Gy showed that the changes in tumor radiosensitivity observed using cell survival endpoints also occurred in tumors left in situ after irradiation with a radiation dose similar to those used in some clinical trials.

Effects of hyperbaric oxygen and a perfluorooctylbromide emulsion on the radiation responses of tumors and normal tissues in rodents

Perfluorochemical emulsions are being examined in many laboratory and clinical studies as possible adjuncts to radiotherapy and chemotherapy. The studies reported here examine the clinical potential of hyperbaric oxygen (HBO) in combination with a highly concentrated perfluorochemical emulsion (Oxygent) containing 100% w/v perfluorooctylbromide (PFOB). HBO alone produced only a small improvement in the radiation response of BA1112 tumors in WAG/rij rats, while regimens combining HBO with Oxygent produced much greater radiation sensitization. A sham emulsion, formulated without the O2-carrying PFOB, did not alter the radiation response of the tumors in comparison with that seen with HBO alone. Neither HBO nor Oxygent plus HBO altered the radiosensitivity of bone marrow progenitor cells in BALB/c mice. HBO alone augmented skin reactions in BALB/c mice, but addition of Oxygent did not alter the skin reactions in comparison to those seen with HBO alone. Regimens combining Oxygent with HBO selectively increased the radiation sensitivity of tumors relative to normal tissues, thereby enhancing the therapeutic ratio. These results support the potential usefulness of perfluorochemical emulsions and HBO in clinical radiation therapy.

Preclinical evaluation of Oxygent as an adjunct to radiotherapy.

These studies examine the potential value of a concentrated emulsion of perfluorooctylbromide (perflubron; Oxygent, Alliance Pharmaceutical Corp.) as an adjunct to radiotherapy. The effects of Oxygent on solid tumors were examined using EMT6 mammary tumors in BALB/c mice and BA1112 rhabdomyosarcomas in WAG/rij rats. Treatment with Oxygent plus O2, carbogen (95% O2/5% CO2), or hyperbaric oxygen (HBO) increased the effects of radiation on the tumors. Analyses of tumor cell survival curves and measurements of intratumor pO2 showed that this potentiation reflected an increase in the proportion of well-oxygenated tumor cells. Neither treatment of the animals with carbogen, O2, or HBO alone nor treatment of air-breathing rodents with Oxygent produced changes of similar magnitude. Treatment with a vehicle emulsion containing all the components of Oxygent except the perflubron did not alter tumor radiosensitivity, showing that tumor radiosensitization required the oxygen-transporting perfluorocarbon, and did not result from any biologic or physiologic effects of other components of the emulsion. These studies also examined the effects of Oxygent on the radiation responses of mouse skin and bone marrow. Oxygent selectively increased the radiation sensitivity of tumors relative to these normal tissues, thereby increasing the therapeutic ratio and producing therapeutic gain. Oxygent appears to warrant further testing as an adjunct to cancer therapy.

Development of a shielded 241Am applicator for continuous low dose rate irradiation of rat rectum

Abstract This paper describes the development of sources, applicators, and techniques that can be used to irradiate rat rectums with continuous irradiation at dose rates of interest in brachytherapy, either with the full circumference of the rectum irradiated, or with half of the circumference shielded from the radiation. The system uses encapsulated 241 Am sources, to irradiate rat rectum with 60 keV photons continuously at a dose rate of up to 50 cGy/hr. Details of the design and fabrication of the 241 Am sources, the rectal applicator, the dosimetry of the system, and the protocols for preparing and irradiating the rats, and for detecting early rectal injury using histological examination of irradiated rectum are presented. Highly effective shielding (attenuation factors as low as 0.04) of half of the circumference of the rat rectum was achievable. Unidirectional 241 Am irradiators for intracavitary brachytherapy offer a unique tool for examining the effects of shielding a portion of the circumference of the rat radiation tolerance of the rectum.

Effects of the Perfluorochemical Emulsion FMIQ on the Radiation Response of EMT6 Tumours

The effects of FMIQ, a perfluorochemical emulsion based on perfluoro-N-methyldecahydroisoquinoline, were examined using BALB/c mice and EMT6 mammary carcinomas. The radiobiological effects of FMIQ were similar to those found previously for Fluosol in the same tumour/host system. Although the perfluorochemical content (20% w/v) and oxygen-carrying capacity of FMIQ are similar to those of Fluosol, the formulation of FMIQ offers some advantages over that of Fluosol. For example, FMIQ has greater stability during storage. FMIQ also is formulated without pluronic F-68 and is based on a perfluorochemical (FMIQ) having a shorter tissue dwell time than the perfluorotripropylamine in Fluosol; it therefore may produce fewer side-effects than Fluosol. The lifetime of the circulating perfluorochemical droplets in BALB/c mice was longer than FMIQ than for Fluosol; this could offer an advantage in fractionated radiotherapy. These findings give reason to expect that FMIQ may prove to be a better emulsion than Fluosol for clinical use as an adjunct to cancer therapy.

Radiation enhancement of lung nodule formation in mice is not potentiated by treatment with a perfluorochemical emulsion and carbogen

The ability of intravenously-injected mouse mammary tumor cells to form lung tumors is increased by irradiation of the thorax 24 h previously. We examined the effects of treatment with a perfluorochemical emulsion (Fluosol-DA, 20%) plus carbogen before and during irradiation on the radiation-induced enhancement of lung nodule formation. We found no evidence that treatment with Fluosol plus carbogen altered the development of tumor nodules in irradiated mouse lungs.

Activity of C-7 Substituted Cyclic Acetal Derivatives of Mitomycin C and Porfiromycin Against Hypoxic and Oxygenated EMT6 Carcinoma Cells In Vitro and In Vivo

A series of cyclic acetal derivatives of mitomycin C (MC) and porfiromycin (POR) were tested for their ability to kill hypoxic and oxygenated EMT6 tumor cells. Amino methyl acetal and thioacetal substitutions at C-7 of MC and POR dramatically increased the cytotoxicity of the compounds to hypoxic EMT6 tumor cells in vitro but had little effect on the aerobic toxicities. In contrast, a methyl substitution at N1a markedly decreased the aerobic cytotoxicities of the compounds but did not alter the hypoxic cytotoxicities. The POR acetal, BMY-42355, had the largest differential between hypoxic and aerobic cytotoxicities yet observed among MC analogs. Preliminary studies in mice showed that BMY-42355 had good antineoplastic activity when used alone or in combination with radiation and was less toxic than POR; the therapeutic ratio of this compound in these initial studies was higher than those of either MC or POR.

Cytotoxicity of BMS-181174: Effects of hypoxia, dicoumarol, and repair deficits

The mitomycin C (MC) analog BMS-181174 (previously designated as BMY25067) has been shown to be active against a variety of solid tumors in mice. The activity of this compound against tumor cell lines resistant to MC and the different toxicity profiles of BMS-181174 and MC suggested that there may be significant differences in the metabolism and the mechanisms of action of these two compounds. Our studies with a mouse mammary tumor cell line (EMT6), a wild-type Chinese hamster cell line (AA8), and three repair-deficient Chinese hamster cell lines (UV4, UV5, and EM9) supported this concept. BMS-181174 was more toxic to all five cell lines in air than in hypoxia; in contrast, MC is more toxic in hypoxia. Dicoumarol (which increases the cytotoxicity of MC in hypoxia and reduces the cytotoxicity of this drug in air) did not alter the cytotoxicity of BMS-181174. This finding suggests that neither DT-diaphorase nor cytochrome b5 reductase is involved in the activation of BMS-181174. Studies with the repair-deficient cell lines suggest that DNA strand breaks are not important to the cytotoxicity of BMS-181174, and that cross-links and adducts may be the critical lesions; these studies also suggest that the lethal lesions produced by BMS-181174 are the same under aerobic and hypoxic conditions.