2005 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? Age-related cataract is a major public health problem. Worldwide it is the major cause of preventable blindness. In the United States (US) approximately 20 and 50 percent of persons 65-74 and 75 years of age, respectively, have age-related cataract that results in visual impairment. Cataract appears in the posterior subcapsular, cortical, and nuclear portions of the eye lens. Over $6 billion is spent each year on cataract-related expenses. Delay of the onset or progress of cataract are important public health concerns, since over 1.2 million eye-lens extractions are performed annually in the US. Costs associated with cataract extractions in the US account for the largest line item, 12 percent of the total Medicare budget. It is estimated that delaying cataract by only 10 years would diminish by 50 percent the number of cataract extractions required. Age-related maculopathy is a blinding condition involving degeneration of the retina’s macula that is responsible for central vision. There is no surgical cure or treatment for this blinding condition, which afflicts approximately 6 percent of the elderly, nor is there a clear path toward a cure. The compromises in quality of life of our elderly and the burden to our health care budgets provide major incentives to explore the etiology of age-related cataract and age-related macular degeneration, as well as to discover means to delay the onset and/or progress of these blinding diseases. The Laboratory for Nutrition and Vision Research (LVNR) is coupling investigations into the etiologies of these debilities with assessments to determine the extent to which diets rich in certain foods or nutrients can offer protection against these vision dysfunctions. Research within this CRIS falls under the National Program 107 - Human Nutrition program components 4: Nutrient requirements, and 6: Prevention of obesity and disease: Relationship between diet, genetics, and lifestyle, and disease. This work should be of interest to the public; ophthalmologists who at present have no cure for age-related macular degeneration (AMD); physicians; nutritionists; gerontologists; public health professionals; biochemists with interests in mechanisms of age-related dysfunction; and those involved in the economics of age-related disability. 2.List the milestones (indicators of progress) from your Project Plan. Objective 1: To determine foods and nutrients that offer protection against onset or progress of age-related cataract and age-related macular degeneration in humans. Objective 2: To determine: how the lens and retina pigmented epithelial layer are constructed, what proteolytic mechanisms are involved in assembly and maintenance of these critical cell layers during aging or stress, and how nutrients and diet can be exploited in order to prolong the functional life of proteases. Objective 3: To establish animal models to determine foods and nutrients that offer protection against onset, or progress, of age-related cataract and age-related macular degeneration, and that also allow for investigations into the mechanisms of these debilities. Milestones GLOSSARY: Prevalence refers to data (cataract or age-related macular degeneration grades) for a cohort, which is gathered at the start of the study. Progress refers to the change of the grades between the start of the study and a later date when the cohort was reexamined. Incidence refers to the number of new cases of the trait which appear at the time of the reexamination but which were not present at the initial baseline examination. Year 2005: 1. Determine how progress of cataract is related to antioxidant intake in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 2. Determine how prevalence of cataract is related to fat intake in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 3. Determine how prevalence of cataract is related to glycemic index/glycemic load in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 4. Determine how prevalence of age-related macular degeneration is related to glycemic index/glycemic load in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 5. Determine how age-related macular degeneration and alcohol intake are related in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 6. Gather data on how ubiquitin and ubiquitin conjugating enzyme 3 direct the lens and retina cell cycle (Objective 2). 7. Gather data regarding how ubiquitin and ubiquitin conjugating enzyme 7 direct the lens and retina cell cycle (Objective 2). 8. Set up models of oxidative damage and ubiquitin dependent proteolytic pathway-dependent degradation (Objective 2). 9. Establish the Osteogenic Disorder Syndrome (ODS) rat as a new animal model for cataract. This rat cannot make vitamin C; therefore it is very useful for antioxidant studies. We will determine levels of carbohydrate that are required to induce cataract at specific rates in these animals (Objective 3). 10. Breed Emory mice for use in experiments to determine the effects of nutrients on age-related cataract (Objective 3). The Emory mouse model developed at the HNRCA is the only laboratory animal that develops age-related cataract at a fixed rate. Year 2006: 1. Determine how progress of cataract is related to fat intake in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 2. Determine how incidence and progress of cataract is related to glycemic index/glycemic load in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 3. Determine how incidence and/or progress of age-related macular degeneration is related to dietary glycemic index/glycemic load in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 4. Determine if ubiquitin conjugating enzymes change location during various phases of the cell cycle (Objective 2). 5. Determine binding/function partners of ubiquitin conjugating enzymes 3 and 7 during various phases of the cell cycle (Objective 2). 6. Determine if inhibition or inactivation of the ubiquitin dependent proteolytic pathway leads to accumulation of protein aggregates using previously established models of oxidative damage which alter ubiquitin dependent proteolytic pathway activity (Objective 2). 7. Establish the ODS (vitamin C-deficient) rat as a new animal model for cataract. Test interventions using carotenoids, specifically zeaxanthin (Objective 3). 8. When sufficient Emory mice are available, use in experiments to determine the effects of zeaxanthin on age-related cataract (Objective 3). Year 2007: 1. Determine how prevalence for cataract is related to diet patterns in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 2. Determine how progress of age-related macular degeneration is related to diet patterns in Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 3. Determine how prevalence of cataract is related to glycemic index/glycemic load in the Age-Related Eye Diseases Study ophthalmologic and nutritional data sets (Objective 1). 4. Determine how prevalence of age-related macular degeneration is related to glycemic index/glycemic load in the Age-Related Eye Diseases Study ophthalmologic and nutritional data sets (Objective 1). 5. Simulate/measure age-related damage to ubiquitin dependent proteolytic pathway (Objective 2). 6. Determine how/that ubiquitin dependent proteolytic pathway selectively targets and degrades oxidized proteins (Objective 2). 7. Determine if ubiquitin protein aggregates are cytotoxic(Objective 2). 8. Establish the ODS (vitamin C-deficient) rat as a new animal model for cataract. Test interventions including vitamins E and lutein (Objective 3). Year 2008: 1 Determine how incidence and progress of cataract is related to intake of antioxidants in the Age-Related Eye Diseases Study ophthalmologic and nutritional data sets (Objective 1). 2. Determine if incidence and progress of age-related macular degeneration is related to glycemic index/glycemic load in the Age-Related Eye Diseases Study ophthalmologic and nutritional data sets (Objective 1). 3. Determine how prevalence for cataract is related to diet patterns in the Framingham ophthalmologic and nutritional data sets (Objective 1). 4. Determine how the prevalence of cataract is related to intake of antioxidants in Framingham ophthalmologic and nutritional data sets is related to glycemic index/glycemic load. (Objective 1) 5. Analyze/evaluate data regarding cataract and alcohol in Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 6 Analyze/evaluate data summary regarding age-related macular degeneration and glycemic index/glycemic load from Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 7. Analyze/evaluate data summary for diet patterns and age-related macular degeneration and glycemic index/glycemic load from Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 8 Analyze/evaluate data summary for diet patterns and cataract and glycemic index/glycemic load from Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 9. Analyze/evaluate data summary regarding relations between age-related macular degeneration and alcohol intake in the Nutrition and Vision Project ophthalmologic and nutritional data sets (Objective 1). 10 Analyze/evaluate data summary age-related macular degeneration and glycemic index/ glycemic load from Age-Related Eye Diseases Study ophthalmologic and nutritional data sets (Objective 1). 11. Simulate/measure age-related damage to ubiquitin dependent proteolytic pathway (Objective 2). 12. Determine how the ubiquitin dependent proteolytic pathway selectively targets and degrades oxidized proteins (Objective 2). 13. Analyze/summarize data - determine how/if we can harness the ubiquitin proteolytic pathway to prolong lens function (Objective 2). 14. Establish the ODS (vitamin C-deficient) rat as a new animal model for cataract. Test interventions with antioxidants and calorie restriction as means to delay cataract (Objective 3). 15. Analyze/evaluate data regarding zeaxanthin interventions for delay of cataract in Emory mice (Objective 3). Year 2009: 1. Analyze/summarize data regarding incidence and progress of age-related macula degeneration in relation to glycemic index/glycemic load from Age-Related Eye Diseases Study ophthalmologic and nutritional data sets (Objective 1). 2. Analyze/summarize data regarding prevalence of age-related macular degeneration and glycemic index/glycemic load from the Framingham ophthalmologic and nutritional data sets (Objective 1). 3. Analyze/summarize data - determine how/if we can harness the ubiquitin proteolytic pathway to prolong retina function (Objective 2). 4. Analyze/summarize data regarding interventions to delay cataract in the ODS (vitamin C-deficient) rat (Objective 3). 5. Analyze/evaluate data regarding zeaxanthin interventions for delay of cataract in Emory mice (Objective 3). 4a.What was the single most significant accomplishment this past year? Establishing mechanisms for lens and retina formation and maintenance of integrity during stress and aging. The accumulation of damaged proteins is partly responsible for cataract formation, age-related macular degeneration (AMD), and many age-related diseases. LNVR scientists are investigating roles for the ubiquitin system, a major protein editing system, in these diseases. To test hypotheses that the ubiquitin system is involved in the removal of damaged proteins, researchers made a mutant ubiquitin. “Normal” ubiquitin forms complexes that can be degraded, thus averting complications or disease. However, complexes that are formed with the mutant ubiquitin cannot. Using the mutant ubiquitin scientists demonstrated that the ubiquitin proteolytic pathway selectively recognizes oxidized proteins. Cells, which incorporated the mutant ubiquitin were far more susceptible to stresses similar to those which humans face upon aging. We also showed that a compromise in the degradation of the ubiquitin conjugates leads to the protein insolubilization that occurs in cataract. We corroborated data that emphasizes the need to define and retain protein quality control to prolong visual function. This is probably achievable through dietary means. We found that the ubiquitin mutants we prepared showed previously undocumented ways that ubiquitin is attached to proteins. Some of these have now been confirmed, and they have phenotypes. This research is leading to completely novel ways of understanding ubiquitin-related aspects of cellular physiology. 4b.List other significant accomplishments, if any. Determining optimal nutrient intake to prolong visual function. With collaborators at the National Institutes of Health, Boston University/Framingham Study, in the HNRCA Nutritional Epidemiology Program (CRIS project 1950-51530-007-01A) and at Harvard University, this laboratory is leading investigations that seek to determine whether and how nutrient intake is related to risk for onset and progression of cataract and age-related maculopathy. Scientists gathered lens and retina images and obtained access to Nurses’ Health Study nutrient intake and personal lifestyle databases. They also obtained ophthalmologic and nutritional information from the Age Related Eye Diseases Study and have started evaluation of data from the Framingham study cohort. Analysis of these large databases is beginning. We found that moderate intake of alcoholic beverages is related to risk for cortical and nuclear early lens opacities. Specifically, moderate wine intake provided diminished risk for age-related lens cortical opacities. Consuming higher levels of fruits and whole grains also provided diminished risk for cataract. Posterior subcapsular cataract and abdominal adiposity. In another collaboration with the Nutritional Epidemiology Program, scientists showed that the chance of developing posterior subcapsular cataract are lower in persons with higher abdominal adiposity and observed that eating according to the recommendations of the Dietary Guidelines for Americans is associated with diminished risk for nuclear cataract. These results emphasize the value of the Dietary Guidelines as a nutritional guide. Glutathione, vitamins C and E and oxidative stress. We observed interactions between glutathione, vitamins C and E that are different and particularly important and obvious when cells are oxidatively stressed. These results emphasize the need to understand biological synergies between nutrients. Oxidative stress, protein quality control and proteolytic function. We pursued the relationships between oxidative stress, protein quality control and proteolytic function in efforts to determine the mechanisms that may exist that allow cells to function longer when nutrition is optimized. These results suggest that antioxidants may prolong the function of the cellular quality control machinery and emphasize the role of nutrition in healthy cell function. 4c.List any significant activities that support special target populations. None. 4d.Progress report. None. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. Research conducted within the Nutrition, Aging and Visual Function CRIS define and describe the cellular protein quality control capabilities ( Milestone 8 - Oxidative damage and ubiquitin dependent proteolytic pathway-dependent degradation). By constructing and investigating the effects of ubiquitin mutants we are revealing previously unknown cellular systems for targeting proteins for various cellular fates and functions. The investigations indicate that the protein quality control mechanisms are compromised by the oxidative stresses, and that these compromises may be related to cataract and age-related macular degeneration (AMD) and most protein precipitation diseases. Optimal nutrition can extend control of the quality control machinery (Milestone 2 - Cataract and fats; 3 – Cataract and glycemic index/glycemic load; 5 – AMD and alcohol; and 4 – AMD and glycemic index/glycemic load). Furthermore, not only can the prevalence of cataract and age-related maculopathy be diminished by appropriate nutrition early in life, but also, the amount of public health resources committed to their treatment can be contained if people follow optimal diets for prolonging visual function. Data from our investigations should be considered when the Dietary Reference Intakes (DRIs) for antioxidant nutrients are reevaluated. This work is related to National Program 107 – Human Nutrition program component 6. Prevention of Obesity and Disease: Relationship between Diet, Genetics, and Lifestyle and to Performance Measure 4.1.2 Improve Human Health by Better Understanding the Nutrient Requirements of Individuals and the Nutritional Value of Foods. 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? The Laboratory for Nutrition and Vision Research made available to the nutrient/food industry estimates of what foods, groups of foods, and supplements should contain as a means to optimize protection of the eye during aging. We are in the process of evaluating dietary carbohydrate content with respect to risk for cataract and AMD. Investigators have adapted modern techniques in proteomics and RNAi technologies that are now available for use by HNRCA scientists. We also have available mutant ubiquitins, which can diminish progress of secondary cataract and cancer progress. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). A Pathway to Clearer Vision, Agricultural Research, August 2005, p. 13. You See How You Eat: How Carbohydrate Intake is Related to Risk for AMD and Cataract. November 2004. Johnson and Johnson Focused Giving Scientific Symposium. Princeton, NJ. Ubiquitin Functions in the Eye: Control of Protein Quality and Cell Cycle. December 2004. NIH. Bethesda, MD. US-Indo Thought Leader Workshop on Collaborative Research. February 2005. Association for Research in Vision and Ophthalmology, National Eye Institute. Madurai, India and Chennai, India. The Function of the Ubiquitin Proteolytic Pathway in Control of Protein Quality, Cell Proliferation and Differentiation. February 2005. 2nd Annual Singapore Eye Research Institute Symposium. Association for Research in Vision and Ophthalmology. Singapore. Ubiquitin Pathway: Roles in Protein Quality Control and Cell Cycle, in Lens and Retina. March 2005. Harvard University. Schepens Eye Research Institute. Ubiquitin-Containing Protein Aggregates in Lipofuscin-Treated Light-Exposed RPE. May 2005. Association for Research in Vision and Ophthalmology. Fort Lauderdale, FL. Glycation of Ubiquitin May Underlie the Molecular Mechanism of Diabetic Cataract. May 2005. Association for Research in Vision and Ophthalmology. Fort Lauderdale, FL. Impairment of Ubiquitin-Proteasome Pathway in Retinal Pigment Epithelial Cells Enhances Expression and Secretion of Vascular Endothelial Growth Factor. May 2005. Association for Research in Vision and Ophthalmology. Fort Lauderdale, FL. Dietary Glycemic Index and Carbohydrate Intake in Relation to Early Age-related Maculopathy. May 2005. Association for Research in Vision and Ophthalmology. Fort Lauderdale, FL. GSH-Modified GammaC-Crystallin is Selectively Degraded by the Ubiquitin-Proteasome Pathway. May 2005. Association for Research in Vision and Ophthalmology. Fort Lauderdale, FL. Ubiquitin Mutants Compromise Cellular Response to Oxidative Stress. June 2005. National Cancer Institute, NIH. Fredricksburg, MD. Roles for the Ubiquitin Pathway in Protein Quality Control Responses to Oxidative Stress in Lens and Retina. June 2005. Meeting on Visual Function: Insights from the Revolution in Biology at the Molecular Level. Tel Aviv, Israel. Microarray Expression Analysis and Statistical Methods Comparison for Caloric Restriction in Emory Mouse. June 2005. Virginia Polytechnic Institute and State University, Blacksburg, VA. Review Publications Taylor, A., Gallagher, M., Shang, F., Boulton, M., Jarrett, S., Rozanowski, B., Rozanowski, M. 2005. Ubiquitin-containing protein aggregates in lipofuscin-treated light-exposed RPE (retinal pigmented epithelial cells). Association of Research and Vision and Opthalmology. Paper No. 1602/B371. Zhang, X., Marques, C., Gallagher, M., Taylor, A., Pereira, P., Shang, F. 2005. Glycation of ubiquitin may underlie the molecular mechanism of diabetic cataract. Association of Research and Vision and Opthalmology. Paper no. 2888/B441. Shang, F., Guo, W., Marques, C., Taylor, A., Pereira, P. 2005. Dual functions of ubiquitin-proteasome pathway in lens protein quality control [abstract]. Association of Research and Vision and Opthalmology. Paper No. 3849/B207. Guo, W., Gallagher, M., Taylor, A., Shang, F. 2005. Impairment of ubiquitin-proteasome pathway in retinal pigment epithelial cells enhances expression and secretion of vascular endothelial growth factor [abstract]. Association of Research and Vision and Opthalmology. Paper No. 3109/B662. Chiu, C., Hubbard, L., Armstrong, J., Rogers, G., Jacques, P.F., Hankinson, S., Willett, W., Chylack, L.T., Taylor, A. 2005. Dietary glycemic index and carbohydrate intake in relation to early age-related maculopathy [abstract]. Association for Research in Vision and Ophthalmology. Paper No. 3300/B853. Zetterberg, M., Taylor, A., Liang, J.N., Shang, F. 2005. GSH-modified gammaC-crystallin is selectively degraded by the ubiquitin-proteasome pathway [abstract]. Investigative Ophthalmology and Visual Science. 46:E-abstract 3891. Paper No. 3891-B249. Chiu, C., Rogers, G., Morris, M., Jacques, P.F., Chylack, L.T., Tung, W., Hankinson, S.E., Willett, W.C., Taylor, A. 2005. Carbohydrate intake and glycemic index in relation to the odds of early cortical and nuclear lens opacities. American Journal of Clinical Nutrition. 81(6):1411-1416. Girao, H., Pereira, P., Taylor, A., Shang, F. 2005. Subcellular redistribution of components of the ubiquitin-proteasome pathway during lens differentiation and maturation. Investigative Ophthalmology and Visual Science. 46(4):1386-1392. Shang, F., Deng, G., Liu, Q., Guo, W., Haas, A.L., Crosas, B., Finley, D., Taylor, A. 2005. Lys-6-modified ubiquitin inhibits ubiquitin-dependent protein degradation. Journal of Biological Chemistry. [published online]. 280(21) 20365-20374. Available at http://www.jbc.org/cgi/content/full/280/21/20365 Siegal, M., Chiu, C., Taylor, A. 2005. Antioxidant Status and Risk for Cataract. In: Bendich, A. and Deckelbaum, R.J., Editors. Preventive Nutrition: The Comprehensive Guide for Health Professionals, Third Edition. Totowa, New Jersey: Humana Press. pp.463-504. Taylor, A. 2004. Letters to the Editor: Reply to J Gomez-Ambrosi et al. American Journal of Clinical Nutrition. 79(5):889.