2004 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? This CRIS, which includes the Vitamins and Carcinogenesis Laboratory and the Nutrition and Cancer Biology Laboratory, is designed to further our understanding of how dietary habits, nutritional supplementation, and other nutritional interventions can be used to prevent cancer. New knowledge attained by the efforts of this CRIS can then be translated into public health initiatives that effectively reduce the burden of cancer in our society. To accomplish this goal, we examine the complex roles that diet plays in modifying metabolic and genetic pathways that lead to human carcinogenesis. Our program is presently focused on the carotenoids, retinoids, and nutrients involved in one-carbon metabolism (methionine, choline and the B-vitamins, folate, B12, B6, and B2). These nutrients have drawn considerable attention with regard to their potential for human cancer prevention. This work is designed to determine how these dietary compounds interact with genetic background to modify molecular and signaling pathways that alter the development of common cancers, such as those of the lung, stomach, liver and colorectum. This research also examines how this response is modified by other exogenous factors (e.g., tobacco smoking, alcohol and chemical carcinogen exposures). Several different approaches, encompassing work in animals, cell culture, and humans, are used in this effort. Cancer continues to be the second most common cause of mortality in the United States, as well as in most developed countries. As the incidence of cardiovascular disease has declined, the significance of the burden that cancer puts on our society has become even more important. Most cancers can be considered as a 'degenerative disease of aging', since elder age is a major risk factor for cancer. Thus, the efforts of this CRIS are highly relevant to the elders in our society. However, cancers appear to develop over many years in an environment that promotes their evolution, and therefore dietary interventions designed to prevent cancer are relevant to younger populations as well since it is the dietary habits of young and middle adulthood that appear to determine the likelihood of cancer developing later on. This CRIS is related to two components of the National Program Action Plan 2: Diet, genetics, lifestyle, and the prevention of obesity and disease and 6: Health promoting properties of plant and animal foods. 2.List the milestones (indicators of progress) from your Project Plan. Vitamins and Carcinogenesis: Year 1 (FY 04): a) Conduct a study in mice to examine select biochemical, molecular and histopathologic consequences of folate depletion, and folate depletion combined with deficiencies of vitamins B2, B6, and B12 (animal study #1). b) In conjunction with a collaborator, initiate the development of a genetically engineered mouse that possesses the murine homologue of the common 677 homozygous MTHFR polymorphism in humans. c) With collaborators, initiate a human folate-depletion trial in which biochemical and molecular changes in the colon can be monitored. Year 2 (FY05): a) Analyze the blood and tissue samples from animal study #1, and analyze the data so obtained. b) Complete development of the 677 MTHFR mouse, and conduct experiments to demonstrate that it behaves physiologically similar to the human. c) Initiate studies in two strains of mice genetically-predisposed to colon cancer (apc1638 and smad3 animals) to determine the susceptibility of these strains to modulation of carcinogenesis by folate status (animal studies 2a and 2b) d) Analyze tissue and blood samples from the human depletion trial, and continue to enroll subjects Year 3 (FY06): a) Complete animal studies 2a and 2b (see above) to determine the modulation of colon carcinogenesis in apc1638 and smad3 animals by folate status. Select one of the two strains to proceed with animal study #3. b) Using the strain most sensitive to folate modulation in studies 2a and 2b, examine how the superimposition of vitamins B2, B6, and B12 depletion further modulate colonic carcinogenesis (animal study #3) c) Complete recruitment of human subjects for depletion study, analyze samples and data Year 4 (FY07) a) Analyze samples and data from animal study #3 b) Prepare manuscript on animal study #4 c) Finish analysis of samples from human studies; prepare manuscripts Year 5 (FY08) a) Prepare manuscript on animal study #3 b) Finish analysis of samples from human depletion study; prepare manuscripts Nutrition and Cancer Biology: Year 1 (FY04): a) Complete molecular cloning and characterization of carotene excentric cleavage enzyme-9'10'- monooxygenase and its regulation of carotenoid metabolism (Manuscript in revision). b) Complete our ongoing study to determine the effect of combined antioxidant nutrients (beta-carotene, vitamin E and C) supplementation on retinoic acid receptor-beta and p53 tumor suppressor function in the lungs of smoke-exposed ferrets. c) Complete our ongoing study to determine the chemopreventive effect of retinoid supplementation on chemical carcinogen-induced, alcohol-promoted hepatic (and peripheral tissue including esophagus, stomach and colon) carcinogenesis in a rat model. d) Complete our ongoing study to determine the chemopreventive effect of combined nutrients (9-cis-retinoic acid and 1,25-vitamin D3) supplementation on retinoid signaling, cell cycle progression and apoptosis on chemical carcinogen induced lung carcinogenesis in an AJ mice model. e) Begin our collaboration with the Vitamin and Carcinogenesis Laboratory to determine the effects of cigarette smoke exposure in the ferrets on one-carbon metabolism and pro-carcinogenic genetic events in the colon. Year 2 (FY05): a) Complete our ongoing study to determine the protective effects of lycopene against carcinogen-induced lung cancer in smoke-exposed ferret model. b) Complete our ongoing study to determine the interaction of lycopene with IGF-1/IGFBP-3 signaling in human lung cancer cells. c) Determine the protective effect of lycopene supplementation on alcohol-altered IGF-1/IGFBP-3 signaling in alcohol-fed rats. Year 3 (FY06): a) Complete our ongoing study to determine the interaction of lycopene metabolite (apo-10'-lycopenoid) with IGF-1/IGFBP-3 signaling in human lung cancer cells. b) Complete our ongoing study to determine the protective effects of lycopene metabolite (apo-10'-lycopenoid) against carcinogen-induced lung cancer in smoke-exposed ferret model. c) Complete our collaborative work with the Vitamin and Carcinogenesis Laboratory to determine the effects of cigarette smoke exposure in the ferrets on one-carbon metabolism and pro-carcinogenic genetic events in the colon. Year 4 (FY07): a) Complete our ongoing study to determine the protective effects of lycopene metabolite (apo-10'-lycopenoid) against carcinogen-induced lung cancer in smoke-exposed ferret model. Year 5 (FY08): a) Complete our ongoing study to determine the protective effects of lycopene metabolite (apo-10'-lycopenoid) against carcinogen-induced lung cancer in smoke-exposed ferret model. 3.Milestones: 3A. Milestones for year 1 not yet accomplished: Year 1 (FY 04): a) Conduct a study in mice to examine select biochemical, molecular and histopathologic consequences of folate depletion, and folate depletion combined with deficiencies of vitamins B2, B6, and B12 (animal study #1). b) In conjunction with a collaborator, initiate the development of a genetically engineered mouse that possesses the murine homologue of the common 677 homozygous MTHFR polymorphism in humans. c) With collaborators, initiate a human folate-depletion trial in which biochemical and molecular changes in the colon can be monitored. Animal study #1 is presently underway. 3B. Milestones to be completed in FYs 2005-2007: FY 2005: a) Analyze the blood and tissue samples from animal study #1, and analyze the data so obtained. b) Complete development of the 677 MTHFR mouse, and conduct experiments to demonstrate that it behaves physiologically similar to the human. c) Initiate studies in two strains of mice genetically-predisposed to colon cancer (apc1638 and smad3 animals) to determine the susceptibility of these strains to modulation of carcinogenesis by folate status (animal studies 2a and 2b) d) Analyze tissue and blood samples from the human depletion trial, and continue to enroll subjects FY 2006: e) Complete animal studies 2a and 2b (see above) to determine the modulation of colon carcinogenesis in apc1638 and smad3 animals by folate status. Select one of the two strains to proceed with animal study #3. f) Using the strain most sensitive to folate modulation in studies 2a and 2b, perform a study to examine how the superimposition of vitamins B2, B6, and B12 depletion further modulate colonic carcinogenesis (animal study #3) g) Complete recruitment of human subjects for depletion study, analyze samples and data FY 2007 h) Analyze results of animal study #3 i) Conduct a mouse study with the 677 MTHFR mice and wild-type mice to determine whether biochemical and molecular changes that occur in the colon in response to folate depletion are genotype-specific (animal study #4). Analyze data from this experiment. j) Initiate animal study #5: the MTHFR 677 mice will be crossed with a strain predisposed to cancer (either the apc1638 or smad3 animals) and carcinogenesis will be examined under control conditions vs. those under folate depletion conditions to determine whether the polymorphism, folate status, and cancer have a 3-way interaction which replicates that human situation. 4.What were the most significant accomplishments this past year? This project was recently established following completion of the Office of Scientific Review Quality Review process. Please see report for 1950-51000-056-00D. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. This project was recently established following completion of the Office of Scientific Review Quality Review process. Please see report for 1950-51000-056-00D. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? This project was recently established following completion of the Office of Scientific Review Quality Review process. Please see report for 1950-51000-056-00D. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. This project was recently established following completion of the Office of Scientific Review Quality Review process. Please see report for 1950-51000-056-00D.