1. RCIU Risque cardio-vasculaire chez ladulte EA2193, Université de la Méditerranée INSERM UMR608 Service de néonatologie, AP-HM, Marseille

2. Barker et al, Lancet 1989

3. Growth of 357 boys who later developed coronary heart disease in a cohort of 4630 boys born in Helsinki 0123456789101112 Age (years) -0.25 -0.2 -0.15 -0.1 -0.05 0 0.05 Standard deviation (Z)-score Height Weight BMI Cohort Eriksson et al, 2001 Barker et al, 2005

5. Arterial Blood Pressure in 20-23 Year Old Adults mmHg *

6. The development of hypertension 204060800 120 100 140 160 180 200 || Systolic Pressure (mmHg) Age (years) Low birthweight Normal

7. Improved early nutrition Improved neurodevelopmental outcome Cardio-vascular disease & type 2 diabetes in adulthood

8. Programmation précoce des maladies cardio-vasculaires et métaboliques Hypertension artérielle: – Mécanismes artériels – Mécanismes rénaux – Mécanismes endocrines Résistance à linsuline

9. Pulse wave velocity in adults

10. Systemic Arterial Compliance (C art ) in premature infants Preterm <30wks Term

11. Umbilical Artery: Elastin Content Term newborn Preterm 30 wks (catechin)

12. Umbilical artery elastin content

13. Umbilical artery collagen content

14. Quarters of weight change (g) Flow mediated dilation (%) Singhal et al, Circulation, 2004 Flow-mediated endothelium-dependent dilation (FMD) and early VLBW infant growth

15. Programmation précoce des maladies cardio-vasculaires et métaboliques Hypertension artérielle: – Mécanismes artériels – Mécanismes rénaux – Mécanismes endocrines Résistance à linsuline

17. Material and Methods Birth Weaning Control IUGR OF IUGR+OF Normal diet LP diet Normal dietEarly OF Normal diet - IUGR: maternal low protein diet (LP) - Early postnatal overfeeding (OF): reduction of litter size during the suckling period 4 groups of rats:

18. Birth weight and glomerular number Birth weight (moy +/- SEM) * Glomerular number (moy +/- SEM) * gn Control IUGR OF IUGR+OF Control IUGR *

19. Histomorphometric analysis of postnatal glomerulogenesis in extremely premature infants Glomerulogenesis markedly decreased in extremely premature infants (EPI) –Correlation with GA Active glomerulogenesis absent in longer (> 40D) surviving EPI Rodriguez et al, Pediatr Dev Pathol 2004

20. Systolic blood pressure (SBP) * * * * * * *

21. Control IUGR OF IUGR+OF Proteinuria (mean+/-SD) CreatCl (mean+/-SD) mg/kg/d Renal function at 4 months mL/min/kgBW *

22. IUGR (nephron number reduction) î Glomerular filtration surface aera Glomerular Pressure Brenner et al. Am J Hypertens 1988 Postnatal overfeeding Arterial blood pressure RAS Vascular remodelling Endocrine factors Glomerulosclerosis Single nephron glomerular filtration rate (SNGFR) Proteinuria

23. Renal function at 12 months (males) Control IUGR OF IUGR+OF * CreatCl (mean+/-SD)

24. Systolic blood pressure at 12 months (males) * *

25. Glomerular number (12 months) (mean+/- SE) * * *

26. Glomerular volume (mean+/- SE) * *

27. HNPN C IUGR IUGR+OF Control OF Control

28. Conclusion In this study: IUGR is associated with nephron number reduction, and elevated arterial blood pressure in young adult rats Postnatal overfeeding of IUGR rats accelerates and prolonges such changes, and is responsible for altered glomerular function and structure in aging rats Postnatal overfeeding, per se improves postnatal nephrogenesis and increases durably arterial pressure in adulthood

29. Glomerular volume p < 10 -3 * Control IUGR IUGR+OF OF

30. Conclusion In this study: IUGR is associated with nephron number reduction, and elevated arterial blood pressure in young adult rats Postnatal overfeeding of IUGR rats accelerates and prolonges such changes, and is responsible for altered glomerular function and structure in aging rats Postnatal overfeeding, per se increases durably arterial pressure in adulthood

31. Glucose tolerance Glucose tolerance test (intra-péritonéale injection of glucose, 2g/kg) Control LP BET * * * * *p<0.05

32. Glomerular number (22 months) * * *

33. Environment: - Environment: + Fetal life and early infancy Later development

34. Ureteric bud Epithelial cells (nephrons) Mesenchymal cells (metanephros) C-ret Wnt 11 WT1Midkine Retinol

35. Several genes harbour a dramatic variation of expression between the Substracted Libraries -15 -10 -5 0 5 10 15 20 25 FTH1TPI1CUL4B GlutPerox exon 1+2 GPOXexon1GPOXexon2GPOXexon3 Genes Difference in cycles on the rat SSH products Kidney R-N Placenta R-N On the SSH products, 4 genes present very high variations between IUGR and controls, with differences between the tissues explored: FTH1 is considerably induced in IUGR-Normal placenta SSH TPI1 is considerably induced in IUGR-Normal kidney SSH CUL4B is considerably reduced in Normal-IUGR kidney SSH GPOX is considerably induced in IUGR-Normal SSH, but not the last exon

36. Santé Durée de vie Alimentation Environnement Génétique Epigénétique Style de vie

37. Mécanismes épigénétiques Susceptible dexpliquer une empreinte métabolique Méthylation (inactivation) ou déméthylation (activation) de séquences CpG du DNA ou des histones (acetylation –desacetylation) Mise en évidence par traitement au bisulfite, D-HPLC et RT PCR Les méthyles proviennent de lenvironnement nutritif (maternel) Les métabolites actifs de loxygène induisent une déméthylation

38. Methyl metabolism: major metabolic intermediates, cofactors, dietary sources (adapted from Cooney et al, J Nutr 2002;132:2393S)

39. ADULT DISEASES CARDIORENAL DISEASE HYPERTENSION DIABETES OBESITY …/… EPIGENETIC MECHANISM MATERNAL ENVIRONMENT PLACENTA & FETUS EPIGENETIC MECHANISM

40. Maternal lymphocytes DNA 37-42 wks Preeclampsia IUGR DMR2-IGF2DMR2bis-IGF2 Placenta 8-14 wks 37-42 wks.éclampsie IUGR Igf2 Differentially Methylated Region 2

41. PERTURBATION Nutrition Caloric restriction Environment Experimental/therapeutic drugs Utero-placental circulation Pre-implantation Environment / conditions Mother Placenta ORGAN SYSTEM 0rgan size/ Cellular phenotype Heart Vascular Brain Liver Adrenals Pancreas Kidney Lung Fat Bone Vasculogenis/ Angiogenis PATHOLOGY Cardiorenal disease Hypertension Diabetes Dyslipidemia Obesity Hypercortisolism GH/GF programming Tumors Respiratory Disease Psychiatric disorders Foetus Epigenetic Mechanism ? ? McMillen and Robinson, 2005

42. Longitudinal growth of hospitalized VLBW infants (Ehrenkranz et al, Pediatrics 1999)

43. Cumulative nutritional deficits Energetic deficit Proteins deficit Embleton Pediatrics 2001; 107:270-273

44. Growth in the NICU influences neurodevelopmental and growth outcomes of ELBW infants (18 months) Outcome*Quartile 1Quartile 2Quartile 3Quartile 4 Weight gain (SD) (g/kg/d) 12.0 (2.1)15.6 (0.8)17.8 (0.8)21.2 (2.0) Normal neurol. exam. 70777686 CP2113 6 MDI < 7039373421 PDI < 7035321814 Neurodev. Impairmt 55494129 Weight < 10th perc. 58615146 Length < 10th perc. 47432928 Head circ; < 10th perc. 3118 22 *p<.05Ehrenkranz R et al, Pediatrics 2006

45. Programmation précoce du risque cardio-vasculaire Suivi à long terme des enfants de faible poids de naissance: –PA – microalbuminurie Nutrition postnatale précoce équilibrée –Évitant un retard de croissance extra-utérin –Respectant le potentiel de croissance individuel Allaitement maternel, mesures éducatives concernant la prévention des autres facteurs de risque cardio-vasculaire Perspectives de recherche: –Caractérisation épidémiologique du risque –Optimisation de la nutrition postnatale des enfants de faible PN –Approches préventives pharmacologiques

46. EA 2193 Université Méditerranée, Marseille, France F. Boubred C. Buffat L Tauzin F. Risso C. Oliver U. Simeoni INSERM U709, Paris D. Vaiman H. Jammes INSERM U364, Paris M. Lelièvre-Pégorier INSERM U608, Marseille F. Dignat-George L. Camoin P. Charpiot