2006 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? Why 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 (LNVR) 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,. 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 by year the currently approved milestones (indicators of research progress) 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 of terms: 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 that appear at the time of the reexamination but were not present at the initial baseline examination. 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 and 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 regarding 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. 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). 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). 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 glycemic index/glycemic load in Framingham ophthalmologic and nutritional data sets (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). 2009 1. Analyze/summarize data regarding incidence and progress of age-related macular 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.List the single most significant research accomplishment during FY 2006. Glycemic Index and Age-Related Macular Degeneration (AMD) There is presently no treatment for age-related macular degeneration. Antioxidant therapy is marginally useful, and only for delaying the progress of moderate to advanced disease. It is clearly important to find means to diminish risk for early stage disease. Using data from the Nutrition and Vision Project with colleagues from Harvard University, we observed that persons with high dietary glycemic index are more prone to early stage AMD. This is a novel observation as dietary glycemic index has not previously been related to AMD risk, although there have been correlations between dietary carbohydrate intake and diabetic retinopathy. These data imply that similar biochemistry and etiologic factors may be at play in non-diabetic, individuals as in diabetics, although the presentation of lesions may differ. The data indicate that simple dietary modification, begun early in life, can be employed to delay the onset of this devastating debility and aid in diminishing health care costs, resulting in marked improved life quality of our elders. We also showed that nuclear cataract – found in the center of the lens, and cortical cataract - the most common form of cataract (common in diabetics) found in the lower part of the lens, are positively related to dietary carbohydrate intake. The data also suggest that risk for cataract, a disease which afflicts 50% of our elderly, can be diminished by simple limitation of dietary carbohydrate. Since cortical cataract are related to total carbohydrate intake where as nuclear cataract are related to dietary glycemic index, these data further indicate that the mechanism of formation of these two major forms of cataracts are different and that slightly different etiologic factors are involved in nuclear as opposed to cortical cataract. This information is also of importance in designing therapies for treatment of cataracts. These results are aligned with the National Program 107 - Human Nutrition program components: 1. Composition of Foods, 2. Bioavailability of Nutrients and Food Components, 4. Nutrient Requirements, 5. Health Promoting Properties of Plant and Animal Foods, and 6. Prevention of Obesity and Disease: Relationship between Diet, Genetics and Lifestyle. 4b.List other significant research accomplishment(s), if any. We successfully cloned and expressed the ubiquitin mutant UbK6W in various forms. This reagent is invaluable for a myriad of investigations into roles for the ubiquitin pathway in quality control and other functions. Since the pathway is involved in regulation of cell proliferation, differentiation, responses to stress, signal transduction, etc, this reagent is finding uses in many fields. We have used the reagent to demonstrate that cells which can not execute proper ubiquitination responses are more subject to stress induced damage than cells which can mount such a response. We also demonstrated that lens formation and cell proliferation rely on a functional ubiquitin system and that inhibiting this system results in both delayed progress through cell cycle and attenuated proliferation as well as delayed differentiation in tissue culture models. 4c.List significant activities that support special target populations. None 4d.Progress report. None 5.Describe the major accomplishments to date and their predicted or actual impact. The primary objectives of the LNVR are to determine etiologic factors in cataractogenesis and AMD and to apply this information to determine ways to delay the onset or progress of these debilities. It is estimated that only a 10 year delay in cataract will eliminate the need for 50 percent of cataract extractions performed annually in the US. Even more profound advantage can be obtained from delaying the onset or progress of age-related macular degeneration (AMD) since there is no current treatment for this debility. Research conducted within the Nutrition, Aging and Visual Function CRIS define and describe the cellular protein quality control capabilities (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 most protein precipitation diseases. The prevalence of cataract and age-related maculopathy may be diminished by appropriate nutrition early in life. In addition, the amount of public health resources committed to their treatment can be contained by following optimal diets for prolonging visual function. Our research data should be considered in the reevaluation of the Dietary Reference Intakes (DRIs) for carbohydrates and antioxidant nutrients. This work is related to National Program 107 - Human Nutrition program component 6. Prevention of Obesity and Disease: Relationship between Diet, Genetics, and Lifestyle. 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 have also completed several evaluations of dietary carbohydrate content with respect to risk for cataract and AMD. 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? Scientists have adapted modern techniques in proteomics and RNAi technologies that are now available for use by HNRCA investigators. We also have available mutant ubiquitins, which can diminish progress of secondary cataract and cancer progress. We have also produced transgenic animals, the analysis of which will indicate ubiquitin-dependent phenomena which are involved in lens and retina formation, and eventually, in maintenance and function of these organs. We have identified: levels of nutrients (vitamins C and E, and the carotenoids, lutein and zeaxanthin, dietary carbohydrates, and the quality and quantity measures of dietary carbohydrates that are associated with prolonged retina and lens function. 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). Popular Press: 1.) "Loss of Central Vision with Age May Be Linked to Quality of Dietary Carbohydrates" Tufts University Medical News Today June 7, 2006 2.) "Tufts Expert Recommends Communicating "Nutrient Density" to Consumers" Friedman School of Nutrition Science & Policy: Nutrition Notes March/April 2006 news http://nutrition.tufts.edu/news/notes/2006-05.html#vision 3.) Macular degeneration may be linked to food United Press International June 7, 2006 http://www.upi.com/ConsumerHealthDaily/view.php?StoryID=20060607-121557-8210r 4.) Dietary Carbohydrates Linked to Cataract Formation, Teaching Brief MedPage Today June 7, 2006 By Judith Groch http://www.medpagetoday.com/Surgery/Ophthalmology/tb/3488 5.) Promoting eye and hand care Scripps Howard News Service June 08, 2006 By Lee Bowman http://www.rocklintoday.com/news/templates/health_news.asp?articleid=3463&zoneid=7 6.) JM USDA HNRCA (Allen Taylor and Chung-Jung Chiu): Macular degeneration may be linked to food United Press International June 7, 2006 http://www.upi.com/ConsumerHealthDaily/view.php?StoryID=20060607-121557-8210r 7.) Macular degeneration may be linked to food BOSTON, June 7 (UPI) -- Age-related macular degeneration may be partly dependent on the types of carbohydrates consumed, a U.S. study finds. 8.) NUTRITION (Allen Taylor): Dietary Carbohydrates Linked to Cataract Formation, Teaching Brief MedPage Today June 7, 2006 By Judith Groch http://www.medpagetoday.com/Surgery/Ophthalmology/tb/3488 9.) NUTRITION (Allen Taylor): Promoting eye and hand care Scripps Howard News Service June 08, 2006 By Lee Bowman http://www.rocklintoday.com/news/templates/health_news.asp?articleid=3463&zoneid=7 11.) AARP the Magazine November/December 2005 Navigator section Health Research: Eye Candy 12.) ARS News Service / ARS Magazine August 2, 2005 A Pathway to Clearer Vision By Rosalie Marion Bliss 13.) Macular degeneration may be linked to food United Press International June 7, 2006 14.) Scripps Howard News Service June 8, 2006 Promoting eye and hand care By Lee Bowman Presentations: Feb 2005 ARVO US-Indo Thought Leader Workshop on Collaborative Research Feb 2005 Singapore Eye Research Institute: Lens and retina function: Roles for the ubiquitin pathway, protein quality control, and nutrition Mar 2005 Harvard University, Schepens Eye Research Institute. Ubiquitin pathway: roles in protein quality control and cell cycle, in lens and retina. May 2005 Ubiquitin Containing Protein Aggregates in Lipofuscin Treated Light Exposed RPE - ARVO Glycation of Ubiquitin May Underlie the Molecular Mechanism of Diabetic Cataract - ARVO Impairment of Ubiquitin Proteasome Pathway in Retinal Pigment Epithelial Cells Enhances Expression and Secretion of Vascular Endothelial Growth Factor - ARVO Dietary Glycemic Index and Carbohydrate Intake in Relation to Early Age related Maculopathy- ARVO GSH Modified GammaC Crystallin is Selectively Degraded by the Ubiquitin Proteasome Pathway - ARVO June 2005 Ubiquitin Mutants Compromise Cellular Response to Oxidative Stress -National Cancer Institute, NIH - Fredricksburg MD. June 2005 Israel- Meeting on Visual Function- insights from the revolution in biology at the molecular level- Roles for the ubiquitin pathway in protein quality control responses to oxidative stress in lens and retina June 2005 International Society for Computational Biology- Microarray expression analysis and statistical methods comparison for caloric restriction in Emory mouse Nov 2005 Cooperative Cataract Research Group meeting - Hawaii - Dietary carbohydrate intake and glycemic index in relation to cortical and nuclear lens opacities in the Age-Related Eye Disease Study July 2005 Department of Cell Biology Harvard University Medical School- K6 in Ubiquitin: Essential for recognition of conjugates by the proteasome and for cellular responses to stress. April 2006 National Eye Institute: Guiding International Collaborations May 2006 Case Western University Department of Nutrition- Candy for the Eyes: Protein damage by dietary carbohydrate in the retina and lens of non-diabetics May 2006 Roles of Environment/Nutrition in Eye Disorders in US/Indo Experience: Building International Collaborations Workshop, ARVO ARVO 2006 Presentations: Abstract Title: Ubiquitin-Proteasome Pathway Is an Important Protein Quality Control Mechanism in the Lens Author: F.Shang, M.Zetterberg, X.Zhang, E.J. Dudek, A.Taylor. 1Human Nutrition Res Ctr on Aging, Tufts University, Boston, MA; Department of Ophthalmology, Institute of Clinical Neuroscience, University of Goteborg, Goteborg, Sweden. Abstract Title: Glutathiolation of gC-Crystallin Reveals a Degradation Signal for the Ubiquitin Proteasome Pathway Author: E.J. Dudek, A.Taylor, X.Zhang, B.Liu, J.J. Liang, F.Shang. HNRC, Tufts University, Boston, MA; Ophthalmic Research Center, Brigham & Womens Hospital, Boston, MA. Abstract Title: Dietary Carbohydrate Intake and Glycemic Index in Relation to Age-Related Macular Degeneration in the Age-Related Eye Disease Study Author: C.-J.Chiu, R.C. Milton, G.Gensler, A.Taylor. Human Nutrition Research Ctr, Tufts University, Boston, MA; AREDS Coordinating Center, The EMMES Corporation, Rockville, MD. Abstract Title: The Ubiquitin-Proteasome Pathway in the Lens Is a Target of Oxidative Stress Author: X.Zhang, A.Taylor, F.Shang. Human Nutrition Res Ctr on Aging, Tufts University, Boston, MA. Abstract Title: Dietary Carbohydrate Intake and Glycemic Index in Relation to Cortical and Nuclear Lens Opacities in the Age-Related Eye Disease Study Author: A.Taylor, C.-J.Chiu, R.C. Milton, G.Gensler. Nutrition & Vision Res-USDA-HNRCA, Tufts University, Boston, MA; AREDS Coordinating Center, The EMMES Corporation, Rockville, MD. Abstract Title: Identification of Ubiquitin-Conjugating Enzyme Ubc4 Substrates in Rat Lens Author: J.Zhou, F.Shang, J.DeGnore, A.Taylor. Laboratory for Nutrition & Vision Research, USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA; Tufts University Core Facility, Mass Spectrometry, Tufts University, Boston, MA. Abstract Title: Differential Regulation of IL-8 and MCP-1 Expression by the Ubiquitin-Proteasome Pathway in Retinal Pigment Epithelial Cells Author: A.F. Fernandes, W.Guo, X.Zhang, M.Gallagher, A.Taylor, P.Pereira, F.Shang. 1Human Nutrition Research Center on Aging, Tufts University, Boston, MA; Centre of Ophthalmology, IBILI - Faculty Of Medicine - University of Coimbra, Coimbra, Portugal.