Biological Markers: New Horizons from -omics

1. Biological Markers: New Horizons from -omics Jose M. Ordovas JM-USDA-HNRCA at Tufts Univ. Boston, MA

2. Current and Future Practice of Disease Prevention/Therapy 2004 Public Health Policy/ Diagnosis Global Advice/Therapy Focused Screening Early detection Personalized Prevention/Therapy Therapeutic Monitoring Predisposition circa2010 Based on Classical and New Biomarkers Based on Genetics

5. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

6. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

7. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

8. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

9. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

10. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

11. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

12. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

13. Dietary Fat & Cholesterol Peripheral Tissues Bile Acids + Cholesterol LDL Liver Intestine HDL IDL Chylomicron Remnant VLDL Chylomicron LPL LPL FFA FFA Lipoprotein MetabolismExogenous - Pathway - Endogenous APOE

14. APOE Gene and Plasma Cholesterol

16. APOE Genotype, Glucose and BMI in the Framingham Offspring Study P interaction = 0.031 Elosua et al. Obes Res. 2003 Dec;11:1502-8.

17. Lipoprotein Metabolism Exogenous - Pathway -Endogenous LPL Dietary Fat & Cholesterol Peripheral Tissues Bile Acids + Cholesterol LDL Liver Intestine CHM HDL IDL Chylomicron Remnant VLDL LPL LPL FFA FFA

18. Genotype/Phenotype Associations: Lack of Consistency • Meta-analyses of population-based studies of effect of Gly188Glu, Asp9Asn, Asn291Ser, and Ser447Ter substitutions at the LPL locus on plasma triglycerides. Wittrup: Circulation, Volume 99(22).June 8, 1999.2901-2907

19. LPL, influence of obesity over the phenotypic expression of the genotype Corella and Ordovas. The metabolic syndrome: a crossroad for genotype-phenotype associations in atherosclerosis. Current Atherosclerosis Reports, 2004.

20. Hypertension • Blood lipids • Obesity / Overweight • Diabetes • Glucose Intolerance CVD RISK FACTORS and Prevention Priorities • Non-modifiable • Risk Factors • Age • Sex • Genes • Coronary heart • disease • Stroke • Peripheral vascular • disease • Health, wellbeing End Points Intermediate Risk Factors • Behavioural • Risk Factors • Tobacco • Diet • Physical Activity • Alcohol Socio-economic, Cultural & Environmental Conditions Modernization, Mechanization, Urbanization, Globalization

21. Perilipin function and Gene Structure Hormone sensitive lipase Perilipin Triacylglycerols Exon1 Exon2 Exon3 Exon4 Exon5 Exon6 Exon7 Exon8 Exon9 Perilipin 6209 10171 11482 13041 14995 (T>C) (A>T) (G>A) (A>G) (A>T)

22. Knock-out mice: the effects on body fat accumulation Martinez-Botas J , Nat Genet. 2000 Dec;26(4):474-9

23. Valencia

24. Odds Ratio for obesity according to PLIN genotypes MEN WOMEN Lu et al. Clin Genet 2004: 66: 299–310

25. Combined effect of the PLIN polymorphisms on weight and BMI (Valencia) PLIN1 PLIN4 PLIN5 PLIN6 Obese phenotype Lean phenotype

26. Replication of PLIN Associations Lu et al. Obesity Research, 2004 Lu et al. J Mol Med, 2005

28. Weight reduction in response to a low caloric diet depending on the PLIN (11482G>A ) polymorphism in obese subjects P=0.015 for interaction detween diet and PLIN

29. Current and Future Practice of Disease Prevention/Therapy 2004 Public Health Policy/ Diagnosis Global Advice/Therapy Focused Screening Early detection Personalized Prevention/Therapy Therapeutic Monitoring Predisposition circa2010 Based on Classical and New Biomarkers Based on Genetics

30. Chemometrics and Bioinformatics PHENOTYPE AND DISEASE? ENVIRONMENT Measurement of Biological Interactions at Different Levels of Biomolecular Organisation: Systems Biology CONTROL LEVELS Organelle Cell Tissue Organ Biofluid Organism Transcriptomics Genome Gene Regulation/Expression Metabolism Proteins Metabonomics Proteomics

32. NMR “Farm”

34. TrueMass® Analysis (Lipomics) CLINICAL / HEALTH INFORMATION Phosphatidylcholine PATIENT Phosphatidylserine Phosphatidylethanolamine Cardiolipin Sphingomyelin Phosphatidylinositol LipidExtract Lyso-PLCholine Triglycerides SAMPLE Diglycerides DATABASE Free Fatty Acids Cholesterol Esters Sterols

35. Surveyor® Data Visualization

36. Metabonomics Tools800 MHz Ultrashielded NMR spectrometer AVANCE 800 spectrometer with the UltraShield- UltraStabilized 800 magnet at Bruker Application Lab

37. 900 MHz 1H 1D NMR Spectrum of Human Urine Intensity (proportional to micromolar proton concentration) Nmrradiofrequencyscale in arbitary units

38. Human Population Response Vector Analysis: Multivariate Effect of Statin Treatment 4 101 3 102 103 104 105 2 106 107 108 1 111 112 113 0 114 PC2 117 119 -1 120 Good Responders 121 122 -2 123 124 -3 -4 -10 -8 -6 -4 -2 0 2 4 Poor Responders subjects 124 Strong % increase in TRG Strong % decrease in TRG 111 122 108 121 113 129 123 114 104 107 103 102 105 112 106 128 125 119 101 117 % increase in TRG, LDL, CHOL and HDL (bis sample trajectory) 125 120 128 129 PC1 PHARMACO-METABONOMICS:EACH LINE REPRESENTS AN INDIVIDUALS’ RESPONSE VECTOR CONNECTING PRE-AND POST-TREATMENT

39. Pattern Recognition Analysis:Control and Soya Dosed Humans Systematic metabolic effect of diet

40. The New Horizon Remains Obscured by Fog and Obstacles

42. Meeting the Challenges • Population Studies of appropriate experimental design • Clinical trials of adequate size and quality • Genetic associations • Product-specific clinical trials in subjects selected for specific genetic variants • Proper evaluation of benefit (FDA?) • Address DTC genetic testing issues • Educate service providers (physicians, nutritionists) • Publish high quality research!!