Systems biological and mechanistic modelling of radiation-induced cancer.

This paper summarises the five presentations at the First International Workshop on Systems Radiation Biology that were concerned with mechanistic models for carcinogenesis. The mathematical description of various hypotheses about the carcinogenic process, and its comparison with available data is an example of systems biology. It promises better understanding of effects at the whole body level based on properties of cells and signalling mechanisms between them. Of these five presentations, three dealt with multistage carcinogenesis within the framework of stochastic multistage clonal expansion models, another presented a deterministic multistage model incorporating chromosomal aberrations and neoplastic transformation, and the last presented a model of DNA double-strand break repair pathways for second breast cancers following radiation therapy.

A generalized Luria-Delbrück model.

We develop extensions of the Luria-Delbrück model that explicitly consider non-exponential growth of normal cells and a birth-death process with mean exponential or Gompertz growth of mutants. Death of mutant cells can be important in clones arising during cancer progression. The use of a birth-death process for growth of mutant cells, as opposed to a pure birth process as in previous work on the Luria-Delbrück model, leads to a large increase in the extra Poisson variation in the size of the mutant cell populations, which needs to be addressed in statistical analyses. We also discuss connections with previous work on carcinogenesis models.

From mechanisms to risk estimation--bridging the chasm.

We have a considerable amount of work ahead of us to determine the importance of the wealth of new information emerging in the fields of sub-cellular, cellular and tissue biology in order to improve the estimation of radiation risk at low dose and protracted dose-rate. In this paper, we suggest that there is a need to develop models of the specific health effects of interest (e.g., carcinogenesis in specific tissues), which embody as much of the mechanistic (i.e., biological) information as is deemed necessary. Although it is not realistic to expect that every radiation-induced process should or could be included, we can hope that the major factors that shape the time dependence of evolution of damage can be identified and quantified to the point where reasonable estimations of risk can be made. Regarding carcinogenesis in particular, the structure of the model itself plays a role in determining the relative importance of various processes. We use a specific form of a multi-stage carcinogenic model to illustrate this point. We show in a review of the application of this model to lung cancer incidence and mortality in two exposed populations that for both high- and low-LET radiation, there is evidence of an "inverse dose-rate" or protraction effect. This result could be of some considerable importance, because it would imply that risk from protracted exposure even to low-LET radiation might be greater than from acute exposure, an opinion not currently held in the radiation protection community. This model also allows prediction of the evolution of the risk over the lifetimes of the exposed individuals. One inference is that radiation-induced initiation (i.e., the first cellular carcinogenic event(s) occurring in normal tissue after the passage of the radiation) may not be the driving factor in the risk, but more important may be the effects of the radiation on already-initiated cells in the tissue. Although present throughout the length of the exposure, radiation-induced initiation appears to play a dominating role only very late in life, and only for those individuals who began their exposure early in life. These conclusions are very dependent, of course, on the hypotheses embodied in the initiation-promotion-conversion paradigm of carcinogenesis. We suggest that recently identified processes, such as the "bystander effect", might affect initiation, promotion, and malignant conversion in different ways. Finally, the manner in which the quality of radiation affects these processes must be understood in the context of the mixed high- and low-LET radiations that are found in the space environment. Important directions in critical experiment definition are suggested, including a renewed emphasis on well-designed animal experiments over extended periods of time.

The use of biologically based cancer risk models in radiation epidemiology.

Biologically based risk projection models for radiation carcinogenesis seek to describe the fundamental biological processes involved in neoplastic transformation of somatic cells into malignant cancer cells. A validated biologically based model, whose parameters have a direct biological interpretation, can also be used to extrapolate cancer risks to different exposure conditions with some confidence. In this article biologically based models for radiation carcinogenesis, including the two-stage clonal expansion (TSCE) model and its extensions, are reviewed. The biological and mathematical bases for such models are described, and the implications of key model parameters for cancer risk assessment examined. Specific applications of versions of the TSCE model to important epidemiological datasets are discussed, including the Colorado uranium miners' cohort; a cohort of Chinese tin miners; the lifespan cohort of atomic bomb survivors in Hiroshima and Nagasaki; and a cohort of over 200,000 workers included in the National Dose Registry (NDR) of Canada.

A new perspective of carcinogenesis from protracted high-LET radiation arises from the two-stage clonal expansion model.

When applied to the Colorado Plateau miner population, the two-stage clonal expansion (TSCE) model of radiation carcinogenesis predicts that radiation-induced promotion dominates radiation-induced initiation. Thus, according to the model, at least for alpha-particle radiation from inhaled radon daughters, lung cancer induction over long periods of protracted irradiation appears to be dominated by radiation-induced modification of the proliferation kinetics of already-initiated cells rather than by direct radiation-induced initiation (i.e., mutation) of normal cells. We explore the possible consequences of this result for radiation exposures to space travelers on long missions. Still unknown is the LET dependence of this effect. Speculations of the cause of this phenomenon include the suggestion that modification of cell kinetics is caused by a "bystander" effect, i.e., the traversal of normal cells by alpha particles, followed by the signaling of these cells to nearby initiated cells which then modify their proliferation kinetics.

Biopersistence, fiber length, and cancer risk assessment for inhaled fibers.

We briefly review the evidence that the carcinogenic risk posed by inhaled fibers depends principally on the lung burden of long fibers. We use a deposition clearance model to generate time-dependent lung burdens in rats of a dozen long fibers for various exposure concentrations. Together with a previously estimated potency factor for long fibers, we use the generated lung burdens to estimate risks of lung cancer associated with inhaled fibers in rats. Over a broad range of exposure concentrations, excess risk is a linear function of exposure concentration. Excess risk of lung cancer is also a linear function of weighted half-life for fibers for which the weighted half-life is short compared to the life span of the rat. We propose an approach to estimating human lung cancer risk associated with inhaled fibers from animal studies.

The power of the European Union protocol to test for carcinogenicity of inhaled fibers.

We evaluate the power of a recent protocol proposed by the European Union (EU) for testing the carcinogenicity of inhaled fibers. We assume that every fiber has oncogenic potential determined by its biopersistence. We use a recently estimated potency for the oncogenic potential of fibers together with experimentally determined "weighted" half-lives (WHL) of a dozen fibers to generate simulations of long-term bioassays conducted according to the EU protocol. We analyze these experiments using standard statistical techniques and determine the number of tests that would have yielded significant results. We conclude that the EU protocol will readily detect the carcinogenic effect of long-lived fibers, such as amosite with a WHL of more than 450 days, and usually detect the effect of fibers, such as RCF1a, with WHL of about 40-50 days. However, the EU protocol has very low power to detect effects of short-lived fibers, such as X607, with WHL of about 10 days.

Analysis of a historical cohort of Chinese tin miners with arsenic, radon, cigarette smoke, and pipe smoke exposures using the biologically based two-stage clonal expansion model.

Hazelton, W. D., Luebeck, E. G., Heidenreich, W. F. and Moolgavkar, S. H. Analysis of a Historical Cohort of Chinese Tin Miners with Arsenic, Radon, Cigarette Smoke, and Pipe Smoke Exposures Using the Biologically Based Two-Stage Clonal Expansion Model. Radiat. Res. 156, 78-94 (2001).The two-stage clonal expansion model is used to analyze lung cancer mortality in a cohort of Yunnan tin miners based on individual histories with multiple exposures to arsenic, radon, cigarette smoke, and pipe smoke. Advances in methodology include the use of nested dose-response models for the parameters of the two-stage clonal expansion model, calculation of attributable risks for all exposure combinations, use of both a fixed lag and a gamma distribution to represent the time between generation of the first malignant cell and death from lung cancer, and scaling of biological parameters allowed by parameter identifiability. The cohort consists of 12,011 males working for the Yunnan Tin Corporation, with complete exposure records, who were initially surveyed in 1976 and followed through 1988. Tobacco and arsenic dominate the attributable risk for lung cancer. Of 842 lung cancer deaths, 21.4% are attributable to tobacco alone, 19.7% to a combination of tobacco and arsenic, 15.8% to arsenic alone, 11% to a combination of arsenic and radon, 9.2% to a combination of tobacco and radon, 8.7% to combination of arsenic, tobacco and radon, 5.5% to radon alone, and 8.7% to background. The models indicate that arsenic, radon and tobacco increase cell division, death and malignant conversion of initiated cells, but with significant differences in net cell proliferation rates in response to the different exposures. Smoking a bamboo water pipe or a Chinese long-stem pipe appears to confer less risk than cigarette use, given equivalent tobacco consumption.

Long man-made fibers and lung cancer risk.

We show that available experimental data from long-term oncogenicity experiments in Fisher rats are consistent with the hypothesis that the oncogenic potential of long man-made mineral fibers is determined mainly by their biopersistence. We present analyses of these data within the initiation-promotion-progression paradigm of carcinogenesis. Our method of analysis can take the temporal pattern of the burden of long fibers in the lungs of individual animals into explicit account. For this analysis, the temporal pattern of lung burden for each animal was imputed from the information obtained from sacrificed animals. The data are consistent with the hypothesis that fibers act as initiators in the rat lung. We present an estimate of the dose-dependent initiation parameter that is based on all the available data.

The role of promotion in carcinogenesis from protracted high-LET exposure.

Recent analysis of epidemiological studies using the two-stage clonal expansion (TSCE) model has shown that radiation-induced promotion dominates radiation-induced initiation for protracted exposures to radon. This strong promotion effect (i.e. enhanced proliferation of already-initiated cells) causes a pronounced 'inverse dose-rate effect', but by a mechanism completely different from those usually discussed in this connection. This rather startling result is discussed along with implications to extended space missions that include a significant amount of high-LET radiation. It is suggested that the effect might be caused by a 'Bystander Effect' by which normal cells in the vicinity of initiated cells are hit by alpha particles and send out signals that modify the cell kinetics of the already-initiated clones.

Air pollution and daily mortality in three U.S. counties.

I used generalized additive models to analyze the time-series of daily total nonaccidental and cause-specific (cardiovascular, cerebrovascular, and chronic obstructive pulmonary disease) deaths over the period 1987-1995 in three major U.S. metropolitan areas: Cook County, Los Angeles County, and Maricopa County. In all three counties I had monitoring information on particulate matter [less than/equal to] 10 microm (PM(10)), carbon monoxide, sulfur dioxide, nitrogen dioxide, and ozone. In Los Angeles, monitoring information on particulate matter [less than/equal to] 2.5 microm (PM(2.5)) was available as well. I present the results of both single and multi-pollutant analyses. Air pollution was associated with each of the mortality end points. With respect to the individual components of the pollution mix, the results indicate considerable heterogeneity of air pollution effects in the different geographic locations. In general, the gases, particularly CO, but not ozone, were much more strongly associated with mortality than was particulate matter. This association was particularly striking in Los Angeles County.

Air pollution and hospital admissions for diseases of the circulatory system in three U.S. metropolitan areas.

Generalized additive models were used to analyze the time series of daily hospital admissions for cardiovascular and cerebrovascular diseases over the period of 1987-1995 in three major metropolitan areas--Cook County, IL; Los Angeles County, CA; and Maricopa County, AZ--in the United States. In Cook and Maricopa Counties, admissions information was only available for the elderly (ages 65 and over), while in Los Angeles County, admissions information was available for all ages. In Cook County, daily monitoring information was available on PM10, CO, SO2, NO2, and O3. In Los Angeles and Maricopa Counties, monitoring information was available daily on the gases, and information on PM10 was available every sixth day. In Los Angeles County, information on PM2.5 was also available every sixth day. In Cook and Los Angeles Counties, associations were found between each pollutant, with the exception of O3, and admissions for cardiovascular disease, with the gases showing the strongest associations. In two-pollutant models with PM and one of the gases, the effect of the gases remained stable, while the effect of PM became unstable and insignificant. In Maricopa County, the gases, with the exception of O3, were weakly associated with hospital admissions for cardiovascular disease, while PM was not. In two-pollutant models with two of CO, SO2, and NO2, the pattern of results is heterogeneous in the three counties. In all three counties, only weak evidence of any association between air pollution and cerebrovascular admissions was found.

Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on initiation and promotion of GST-P-positive foci in rat liver: A quantitative analysis of experimental data using a stochastic model.

We use a stochastic model describing initiation and clonal growth of altered cells to analyze data from an initiation-promotion hepatocarcinogenesis experiment in female Wistar rats. Starting at 7 weeks of age, the animals were treated for 10 days with the initiating agent diethylnitrosamine (DEN, 10 mg/kg body wt per day). After a 10-week resting period, the animals were treated either with corn oil or with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) via biweekly sc injections of 1.4 microg/kg body wt of TCDD dissolved in corn oil. Groups of four or five animals were euthanized 3, 17, 31, 73, and 115 days after start of TCDD/corn oil treatment. The data analyzed consist of the number and sizes of GST-P-positive focal transections at various time points. By fitting the model to the data, we estimate the rates of initiation, cell division, and cell death during different time periods of the experiment. The model estimates of cell kinetic parameters are consistent with directly made experimental observations of cell division and cell death. The model predicts that DEN-induced initiation of GST-P-positive cells is highly protracted in controls and TCDD-treated animals alike. We also find that TCDD interferes with the normal rate at which cells with (DEN-inflicted) DNA damage are converted into cells expressing the GST-P-positive phenotype, suggesting a TCDD-mediated "acceleration" of the appearance of de novo GST-P-positive initiated cells from damaged precursor cells. Furthermore, the model predicts a significant reduction in the rate of apoptosis within the first 4 to 5 weeks of TCDD treatment, and after 10 weeks of TCDD treatment, but not in between.

Quantitative analysis of tumor initiation in rat liver: role of cell replication and cell death (apoptosis).

The formation and development of initiated cells has been studied at the beginning of hepatocarcinogenesis. Rats received the genotoxic carcinogen N-nitrosomorpholine (NNM); placental glutathione S-transferase was used as a marker of initiated cells (G+ cells). Single G+ cells appeared within 24 h after NNM; their frequency increased steeply for approximately 2 weeks, then decreased and finally remained constant. G+ foci consisting of >/=2 G+ cells appeared successively after the single cells. Histological determination of DNA replication and apoptosis revealed that: the formation of single G+ cells may not depend on DNA replication of precursor cells; single G+ cells showed considerably lower DNA replication than G- normal hepatocytes; from the 2-cell stage onwards G+ foci displayed enhanced DNA replication and apoptosis. Data from histological sections were transformed into the third dimension by a new stereological method which considers the non-spherical shape of many G+ lesions. Rates of division and death of G+ cells and of formation and growth of G+ foci were estimated by a stochastic model: initially G+ clones appeared at a rate of 12 000 per day and liver until a maximal number of 176 000 (phase I) was reached; thereafter they declined to 134 000 (phase II); they then remained constant (phase III). Estimated division rates of G+ cells decreased from phase I to phase III, while the death rate increased in phase II, when every third G+ clone disappeared. As a result, at day 50 after NNM only 0.3% of G+ single cells had formed a clone containing >/=5 cells. In conclusion, experimental and computed parameters provide direct evidence that hepatocarcinogenesis evolves clonally and that initiated hepatocytes have a selective proliferation advantage, associated with an enhanced potential to undergo apoptosis. Thereby, depending on the conditions, initiated clones expand or become extinct. Extinction may lead to reversion of the biological effects of initiation.

Lung cancer risk associated with exposure to man-made fibers.

We show that available experimental data from long-term experiments are consistent with the hypothesis that the oncogenic potential of man-made fibers is determined completely by their biopersistence. We present an analysis of these data within the initiation-promotion-progression paradigm of carcinogenesis. Our method of analysis takes explicit account of the temporal pattern of fiber burden in the rat lung, and suggests that fibers act as initiators in the lung. We estimate a dose-dependent initiation parameter and show how it can be transported to human populations for assessment of the risk of lung cancer following exposure to man-made fibers.

Air pollution and hospital admissions for chronic obstructive pulmonary disease in three metropolitan areas in the United States.

I used generalized additive models to analyze the time series of daily admissions for chronic obstructive pulmonary disease (COPD) over the period 1987-1995 in three major metropolitan areas, Cook County, Los Angeles County, and Maricopa County, in the United States. In Cook and Maricopa counties I had admissions information only for the elderly (ages 65 yr and over). In Los Angeles County I had admissions information for all ages. In all three counties I had monitoring information on PM10, CO, SO2, NO2, and O3. In Los Angeles County, I had information on PM2.5 in addition. In Cook and Maricopa counties there was weak evidence of an association between the gaseous pollutants other than ozone and admissions. There was no evidence of an association with PM. In Cook and Los Angeles counties, in single-pollutant models, ozone was associated with admissions during the period April-September but not in full-year analyses. In Los Angeles, the other gases were strongly associated with COPD admissions. PM was also associated with admissions in single-pollutant models. The coefficients for PM were greatly attenuated and became insignificant in joint analyses with any one of the gases (with the exception of ozone). Analyses in three broad age ranges (0-19, 20-64, and 65 yr and over) yielded similar results. The results indicate that the gases, other than ozone, were more strongly associated with COPD admissions than PM and that there was considerable heterogeneity in the effects of individual pollutants in different geographic areas of the country.