In memoriam: James A.Miller (1915–2000)

Miriam C. Poirier

Division of Basic Sciences, NCI, National Institutes of Health, Bethesda, MD 20892-4255, USA

Dr James (Jim) Alexander Miller, Professor Emeritus of Oncology at the University of Wisconsin (Madison) McArdle Laboratories for Cancer Research died on December 24, 2000 at the age of 85. His illustrious research career began in 1943, and was conducted between 1945 and 1987 with his first wife and research partner Elizabeth (Betty) Cavert Miller, and between 1987 and 1998 on his own. His research findings provided the foundations for our current knowledge of the metabolic activation and biological disposition of chemical carcinogens. The insights that were gained from the Millers' research provided the basis for much of what we know today as chemical carcinogenesis, and have had profound influences in the areas of toxicology, pharmacology, cancer chemoprevention and the molecular epidemiology of human cancer.

As much as it is a story of scientific brilliance and achievement, the story of Jim's life is a lesson in courage, persistence, hard work and a profound capacity to love. Jim was born in Dormont, Pennsylvania, May 27, 1915, the fifth of six brothers. By 1930 he had lost his mother, a beloved Aunt, and two of his brothers, and his father had become unable to work. As the times were permeated by the Depression, Jim worked in a steel mill after high school and went to night school, before obtaining financial support from the National Youth Administration, part of Roosevelt's New Deal. This funding, as well as help from his older brothers, provided an opportunity to attend the University of Pittsburgh where he obtained a B.S. in Chemistry with the highest honors in 1939. During his undergraduate years he worked part time for Professor Charles G.King, a pioneer in vitamin C research, and developed a passion for research through mentoring by both Dr King and his postdoctoral associate, Dr Max O.Schultze. Dr Schultze convinced Jim to apply for graduate school at his alma mater, the University of Wisconsin. In the fall of 1939 Jim entered the University of Wisconsin (Madison) Department of Biochemistry as a prestigious Wisconsin Alumni Research Foundation (WARF) Scholar in the laboratory of Dr Carl A.Baumann. At the time Dr Baumann was using hydrocarbons and aminoazo dyes to produce tumors in rodents, and Jim studied the metabolism, structure–activity relationships and chemoprevention of these compounds, as well as the fluorescent properties of polycyclic aromatic hydrocarbons. He obtained his PhD in 1943.

In 1942, while working as a teaching assistant for a graduate level course in Biochemistry, Jim met and fell in love with one of his students, Elizabeth Cavert, who was also a WARF Scholar. Betty had graduated from the University of Minnesota with the highest honors, and Jim was impressed with everything about her. Due to the limited opportunities for women at that time, Betty had been shunted into a joint major, including both Biochemistry and Home Economics, and Jim intervened to have Dr Baumann take her as a Biochemistry graduate student. (It would not be the last time that Jim would act as a champion of women's rights.) Jim and Betty were married 8 months after they met and began a collaboration that was long and highly successful in both the personal and professional spheres.

In 1944, based largely on his work with chemical carcinogens, Jim was recruited by the Director, Dr Harold Rusch, to a position as faculty member for the newly established McArdle Laboratory for Cancer Research. When Betty obtained her PhD in 1945, Dr Rusch supported her recruitment as a postdoctoral fellow and ultimately as a faculty member in 1947. Thus, Jim and Betty began what would prove to be 42 years of productive collaboration resulting in the training of at least 50 graduate students and postdoctoral fellows, as well as many laboratory assistants and undergraduate students. They were the joint recipients of multiple prestigious awards, including the Papanicolaou, Bertner, Rosenstiel, FASEB Life Sciences, Bristol-Meyers and General Motors Mott awards. Most of these awards were given jointly, as was their election to the National Academy of Sciences in 1978, their appointments to WARF Professorships in Oncology in 1980, their appointments to Van Rensselaer Potter Professorships in 1982 and their appointments to WARF Senior Distinguished Professors of Oncology in 1984. In addition, there were some individual honors, including Jim's AACR Clowes award in 1969 and Betty's Presidency of the AACR in 1977.

As young investigators the Millers set out to elucidate the process by which a chemical causes cancer, and in doing so reasoned that there must be metabolic pathways whereby chemicals become transformed through one or several steps to reach an `ultimate' carcinogen, hypothesized to damage a critical macromolecule and destroy the cellular growth control. One way they approached this question was by synthesizing multiple analogs of a known carcinogen and testing the carcinogenic activity of each. They were searching for a metabolite more carcinogenic than the parent compound, and the testing of each new chemical in a rodent bioassay took 6–18 months. Their early studies with N,N-dimethyl-4- aminoazobenzene (DAB) yielded only one compound with similar carcinogenic potency, N-methyl-4-aminoazobenzene (MAB). In later years the N-hydroxy metabolite of MAB was synthesized and the metabolic pathways elucidated, but the first break in following the metabolic pathway of a chemical carcinogen to an ultimate carcinogen occurred with the aromatic amine N-acetyl-2-aminofluorene (AAF). The N-hydroxy metabolite of AAF was shown to accumulate in the urine of rodents fed AAF. When the Millers synthesized and tested N-OH-AAF they found that it was more carcinogenic than the parent compound. Subsequent studies identified a reactive sulfuric acid ester of N-OH-AAF as the next metabolite in the activation pathway and an ultimate AAF metabolite responsible for binding directly to nucleic acids and proteins. Investigations in multiple species eventually elucidated a complicated metabolic scheme for AAF metabolism that involved sulfotransferase activity and acetyltransferase activity.

In parallel with the structure–activity approach to understanding the process of chemical carcinogenesis, the Millers pioneered the study of specific enzymes involved in metabolism of carcinogens. Between 1948 and 1957, they conducted a series of experiments that demonstrated oxidative metabolism of xenobiotics involving microsomal enzymes that were eventually identified as endoplasmic reticulum-associated cytochromes P450. Several-fold increases in microsomal enzyme activities responsible for demethylation, reduction and hydroxylation were demonstrated to occur in rat liver within hours after polycyclic aromatic hydrocarbon administration. We now know that these enzymes participate in the inactivation and activation of endogenous substances and foreign compounds, and can lead to both urinary excretion and covalent binding of carcinogens to macromolecules. Over the years the Millers elucidated the metabolic activation pathways of a wide variety of chemicals, including aromatic amines, aminoazo dyes, aflatoxin B1, safrole, estragole and ethyl carbamate.

Early in their investigations with DAB the Millers had become aware that livers from rats fed the compound turned pink when exposed to acid. This test was used routinely to indicate the presence of DAB. After the failure of multiple attempts to separate liver proteins from DAB, the Millers came to the revolutionary conclusion that the carcinogen was bound covalently to liver protein. (As DNA-bound azo dyes did not turn pink with acid, azo dye–DNA adducts were not characterized until much later.) In 1947 the Millers proposed a protein-binding hypothesis of tumor formation. However, subsequent to Watson and Crick's elucidation of the structure of DNA, published in 1953, the Millers began studies to identify DNA-bound products (adducts) of chemical carcinogens. In the 1950s the emergent availability of radiolabeling made possible the determination of DNA adduct formation in experimental systems, and the Millers took full advantage of this opportunity. Their pioneering efforts in characterization of DNA adducts of many different chemical carcinogens contributed to the future development of various methodologies that are used today to measure DNA adduct formation in experimental models and in humans. Some of their most elegant studies to elucidate the relationship between DNA adduct formation and tumorigenesis were conducted in the later years using mice that form liver tumors in the presence of a single DNA adduct induced by a specific carcinogen.

Excellent detailed summaries of the Millers' research findings have been previously published (13) and described in recent obituaries for Jim (4,5). Arguably the Millers' greatest contribution was the painstaking accumulation of evidence, over a span of many years, that most carcinogenic chemicals are metabolized in vivo to electrophilic species that bind to nucleophilic targets on proteins and nucleic acids. By the late 1960s the Millers had accumulated sufficient evidence for this to satisfy even their own highly exacting requirements for proof, and they stated and supported their hypothesis in print in 1969 (6). The presumption that DNA was the critical target for carcinogenesis was further substantiated by pioneering mutagenesis studies in Bacillus subtilis and the Ames salmonella assay, which demonstrated the direct mutagenicity of the activated electrophilic forms of carcinogens of various classes (polycyclic aromatic hydrocarbons, aromatic amines, aflatoxins and alkyl carbamates) while the parent, unmetabolized compounds remained non-mutagenic. Finally, Jim and Betty collaborated in studies showing that in tumors induced by many chemical carcinogens the ras oncogene is mutated by the activated carcinogen in codons critical for neoplastic transformation.

After many years of great scientific and domestic success, Jim was devastated upon Betty's untimely death from kidney cancer in October of 1987. In addition to their productive careers, they had raised two daughters (Linda Miller Forbess and Helen Miller Alexander) who were grown and married, and had twice become grandparents. Much to Jim's credit, however, he was able to rebuild his life with Barbara Butler, who had been a laboratory assistant in the Miller lab from 1965 to 1974, and who became his second wife in December of 1988. Jim and Barbara Miller delighted in traveling, going to scientific meetings and visiting with family and friends. They had a very full life, as Barbara, with Jim's urging, was pursuing her PhD in Near Eastern Studies, and Jim continued to go into McArdle daily. Much of this ended in October of 1998 when Jim was confined to constant care.

Perhaps the least-known aspect of Jim Miller's life was his support for and delight in the accomplishments of others. Maybe because his own education had been obtained with such ... effort, Jim appreciated and empathized with the personal and professional struggles of his family, friends and associates. In his later years he had more time to indulge this desire and followed the careers of his former associates and friends with avid interest. Jim Miller was a great scientist, a man of the highest integrity and a true friend. He will be missed but not forgotten.



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Fig. 1. James and Barbara Miller.

 
References

  1. Miller,E.C. and Miller,J.A. (1981) Searches for ultimate chemical carcinogens and their reactions with cellular macromolecules. Cancer, 47, 2327–2345.[ISI][Medline]
  2. Miller,J.A. (1994) Research in chemical carcinogenesis with Elizabeth Miller—a trail of discovery with our associates. Drug Metab. Rev., 26, 1–36.[ISI][Medline]
  3. Miller,J.A. (1998) The metabolism of xenobiotics to reactive electrophiles in chemical carcinogenesis and mutagenesis: a collaboration with Elizabeth Cavert Miller and our associates. Drug Metab. Rev., 30, 645–674.[ISI][Medline]
  4. Kadlubar,F.F. (2001) James A.Miller (1915–2000) and Elizabeth C.Miller (1920–1987). Chem. Res. Toxicol., in press.
  5. Conney,A. (2001) James A.Miller (1915–2000) and Elizabeth C.Miller (1920–1987). Cancer Res., in press.
  6. Miller,J.A. and Miller,E.C. (1969) Metabolic action of carcinogenic aromatic amines and amides via N-hydroxylation and N-hydroxy esterification and its relationship to ultimate carcinogens as electrophilic reactants. Jerusalem Symp. Quantum Chem. Biochem., 1, 237–261.




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