Michael E. Placke

Development of Inhalational Agents for Oncologic Use

Because regional chemotherapy has been useful in treatment and palliation of many cancer types, the concept of delivering drugs by inhalation for the treatment of cancers in the lung is attractive. Much higher local drug exposure can be achieved with total doses considerably lower than those required for systemic administration, resulting in lower exposure of nonrespiratory tract tissues to potentially toxic drugs. Regional delivery of chemotherapy to the respiratory tract has been shown to have activity in preclinical and clinical studies. Technical improvements in delivery methods have now made it possible to conduct trials of inhalational agents, both to treat cancers affecting the respiratory tract and to deliver other drugs used in cancer patients. This review discusses the rationale of drug delivery by the inhalational route, its technical challenges, preclinical and clinical experiences, limitations, and promise.

Adult peripheral lung organ culture--a model for respiratory tract toxicology.

This report describes procedures to culture 1- to 2-mm-thick cross sections of lung lobes for periods of 4 to 6 weeks. Normal morphologic and macromolecular composition are maintained. Previous attempts to maintain adult peripheral lung cultures for periods beyond 7-10 days or to examine respiratory disorders in vitro other than acute changes have been generally unsuccessful. Eight different, supplemented, serum-free media, mixed with heated liquid agarose were infused into the airways of hamster and rat lungs. Cross sections were explanted onto squares of porous surgical packing material, placed in medium, and incubated for 4 to 6 weeks. The ability of each medium to maintain normal lung was assessed microscopically by quantitative image analysis and by biochemical analyses. The optimal medium formulation for each species is described. The adult peripheral lung culture system may provide toxicologists with a unique model for mechanistic and safety evaluations of potential lung toxicants.

In vitro models of lung toxicity

In vitro assays that emphasize cellular components critical to the host defense system have been developed to evaluate pulmonary toxicity and define deleterious changes in parenchymal cell populations. Assays that employ pulmonary alveolar macrophages (PAM) have demonstrated good correlation between macrophage toxicity and pulmonary fibrogenicity for many inorganic compounds. The PAM assays provide simple and inexpensive screens of potential respiratory tract toxicity. Many investigators screen chemicals for their ability to alter the mucosal epithelial cell conducting airways by performing tracheal organ cultures. The tracheal assays have also provided useful screens for Vitamin A analogues required for epithelial cell differentiation. Most recently, in vitro respiratory tract models have been extended to include whole-lung explants, an approach that allows for development of fibrosis and epithelial cell toxicity after in vitro exposure to inorganic and organic fibrogens. The whole-lung explant system appears to duplicate the in vivo response to a variety of lung toxins, including bleomycin, silica, and crocidolite asbestos. Together, these assays provide a description of potential toxicity to key components of the lung, emphasizing the pulmonary macrophage, conducting airways, and alveolar septae. It is expected that continued research in these models will enhance their predictive abilities and utility in risk assessment.

Chronic inhalation oncogenicity study of isoprene in B6C3F1 mice

The oncogenic potential of isoprene as affected by concentration, length of daily exposure, and weeks of exposure over the life-span of the animal, as independent variables, was evaluated. Ten groups were exposed for 8 h/day, 5 days/week as follows (ppm-weeks): 0-80, 10-80, 70-40, 70-80, 140-40, 280-20, 280-80, 700-80, 2200-40, 2200-80. Two groups were exposed for 4 h/day: 2200-20, 2200-80. Groups were held until 96 or 105 weeks on study. The concentration x time (duration of exposure) values provided a series of theoretically equivalent exposure hazards. There was an exposure-related increased incidence of liver, lung, Harderian gland and forestomach tumors, and hemangiosarcomas and histiocytic sarcomas. The LOEL appeared to be 70 ppm. These results are similar to the profile of tumors seen in 1,3-butadiene (BD)-exposed mice without the early onset of T-cell lymphoma as seen with BD. Isoprene appears to be about one order of magnitude less potent than BD in mice. Statistical analyses indicated that the product of isoprene concentration, and length/duration of exposure was not a sufficient basis for predicting tumor risk at any site. Extrapolation of tumor probability between the high and low doses based on cumulative exposure was not appropriate and could not be justified by statistical models. A threshold effect level and strong nonlinearities with respect to concentration appeared to exist for tumor development in this study.

Considerations in developing successful, population-based molecular screening and prevention of lung cancer

The current mortality rate for lung cancer exceeds 85%, as it has for the last 3 decades. This statistic reflects the utility of the major diagnostic tool that has been used during this period to diagnose lung cancer: the chest X‐ray. The overwhelming majority of new cases of lung cancer that are detected with chest X‐rays involve individuals who already have regional or distant metastatic disease. Because the systemic treatment of this disease has not improved greatly, patients with metastatic disease rarely are cured. This article reviews the issues involved with the development of sputum‐based cellular diagnostics for early stage lung cancer. The biomarker, heterogeneous nuclear ribonucleoprotein A2/B1, is the lead marker for this approach. It has been used in several studies in independent cohorts that have suggested that its overexpression in bronchial epithelial cells is associated highly with the development of lung cancer. This marker is detectable 1 year or more prior to the detection of lung cancer by chest X‐ray. Finding this early airway‐confined phase of lung cancer may allow for the evolution of new management approaches for very early stage lung cancer. Research activities, such aerosolized chemoprevention, are discussed. Cancer 2000;89:2465–7.

Pharmacokinetics of 2′,3′-dideoxyadenosine in dogs

The pharmacokinetics of 2′,3′-dideoxyadenosine (ddAdo) and 2′-3′-dideoxyinosine (ddIno) were determined after intravenous bolus administration and long-term intravenous infusion of ddAdo in dogs. ddAdo was rapidly deaminated to ddIno and ddAdo plasma concentrations were only a fraction of ddIno concentrations. The total body clearance of ddAdo exceeded the literature value for the cardiac output of the dog, indicating an extremely rapid metabolism, and the existence of extrahepatic metabolism. Urinary excretion of unchanged ddAdo was a minor route of elimination (∼ 1%). The pharmacokinetics of ddIno was determined assuming complete conversions of ddAdo to ddIno. ddIno elimination was dose-dependent with total body clearance ranging from 4 to 55 ml/min/kg in individual animals. The plasma half-life was approximately 30 min after most routes of administration, but increased to approximately 60 min in two animals receiving a large intravenous dose of 500 mg/kg. ddIno penetrated into the cerebrospinal fluid to a limited extent, reaching concentrations of 3–11% of those in plasma. Urinary excretion of unchanged ddIno accounted for approximately 20% of the administered dose of ddAdo, while uric acid and hypoxanthine were minor urinary metabolites.Concentrations exceeding the in vitro minimal viral inhibitory concentration (2.4 μg/mL) could be safely maintained in plasma for a 10-day period. Infusions which gave cerebrospinal fluid concentrations of 12 to 17 μLg/mL resulted in dose limiting myelosuppression and intestinal toxicity, after less than 10 days of infusion. Orally administered ddAdo was absorbed as ddIno, with bioavailabilities ranging from 28 to 93% in experiments where no emesis occurred. These studies indicate the rapid in vivo conversion of ddAdo to ddIno, and support the selection of ddIno over ddAdo for further drug development.

Asbestos Ill Peripheral Lunc Culture a Species Comparison of Pulmonary Tissue Response

We have recently developed culture techniques to allow the long-term maintenance of adult peripheral lung tissue in vitro from a variety of mammalian species including hamster, rat, bovine, and human. The technique involves perfusion of the major airways with agarose gel and culture media followed by thin sectioning and culture on porous surgical foam. Cross sections of lung lobes 1-2 mm thick have been cultured for periods beyond four weeks with maintenance of structural and biochemical integrity of the lung. In vitro exposure of lung explants to crocidolite asbestos through the airways produced fibrotic and hyperplastic lesions similar to those reported after in vivo exposure. The incidence and severity of interstitial fibrous was concentration-dependent, including the human specimens, and the morphologic appearance of the lesions was similar in explants derived from each species. The lung explant model is well suited for further mechanistic evaluations of asbestos-induced lung lesions. It is notable that the pulmonary lesions were produced without the possibility for recruitment of hematogenous inflammatory cell populations.

Age-sensitive morphologic changes in tracheal organ culture following carcinogen exposure

This study was designed to assess the differential sensitivity of tracheal explants obtained from female golden Syrian hamsters at 30, 100, or 180 days of age to exposure to crocidolite asbestos or benzo[a]pyrene (BaP). Explants were evaluated at 1 or 4 weeks following the initial exposure. Hyperplasia and squamous metaplasia of the mucosal epithelium were observed in exposed explants. Alcian blue-periodic acid-Schiff base stains demonstrated decreased amounts of intracellular acidic mucopolysaccharides in exposed explants. Morphometry supported the qualitative observations that lesions were more severe in tracheas from 30-day-old animals than lesions from the 100- and 180-day-old donor hamsters. Thus, it appears that the age-dependent, early toxic response of the tracheal organ culture model to carcinogens parallels that of animal models.

Moving to the routine management of pre symptomatic lung cancer.

Lung cancer is the worlds leading cause of cancer death. Since progress in the treatment of this cancer has been exceedingly slow, the upswing in tobacco consumption in many sectors becomes even more tragic. One area for cautious optimism is the recent pilot reports of improved early lung cancer detection using new spiral CT techniques from institutions in Japan and New York. The prospect of improved early detection in a major cancer raises a number of public health concerns and highlights the importance of critical validation of this proposed new tool. From experience with early detection-based management of other cancers, it is evident that the entire process of detection, case validation, intervention, monitoring and public education needs to be carefully developed. The International Association for the Study of Lung Cancer has worked with the National Cancer Institute over the last decade to nurture interest and expertise in conducting population-based management of early lung cancer. A distillation of this process up to the current time is reviewed in this manuscript.