1. Epigenetics, Diet and Disease Giles Elliott

2. Diet Epigenetics DNA Methylation Disease Colon Cancer Overview

4. Retained through mitosisC C C C C C A A A A A A A A G G G G G G G G T T T T T T C C C C C C A A A A A A A A G G G G G G G G T T T T T T Dnmt3 Dnmt1

6. Genomic stability

7. Repression of transposable elements

8. High Density – CpG Islands (CGIs)

9. Located in promoters and introns

11. Due to reduction in global methylation - hypomethylation

12. Increased genomic instability – increased mutation rate

13. Reactivation of transposable elements – chromosomal rearrangements

15. Present in both hereditary and sporadic types of cancer

16. Occurs at all stages of cancer development

17. Interacts with genetic lesions – 2 hit

19. DNA methylation in relation to disease – Colorectal cancer Adenoma Lifetime risk 50% Carcinoma Lifetime risk 5% Healthy Mucosa Somatic mutation….. Dietary and microbiological factors act on the mucosa for decades before the emergence of tumours. APC K-ras p53 Epigenetic deregulation… MLH1 p16 ESR-1 Tumour Biology Diet-Related “Field Effects” ?

20. Evidence for the Field Effect

21. Evidence for the Field Effect Adenoma Carcinoma Healthy Mucosa Diet-Related “Field Effects” ?

22. Evidence for the Field Effect Adenoma Carcinoma Healthy Mucosa Diet-Related “Field Effects” ?

24. 78.9% of cancer patients

25. 87.9% of non- cancer patients

26. informative variables: APC, HPP1, p16, SFRP4, WIF1 and ESR1

27. CGI methylation of SFRP4, SFRP5 and WIF1 was used to correctly identify:

28. 61.5% of adanoma patients

29. 78.9% of neoplasia-free subjects

30. This indicates that apparently normal mucosa of patients with neoplasia has undergone significant epigenetic modification associated with the onset of carcinogenesis.

31. Methylation of the genes selected by the models may play a role in the earliest stages of the development of colorectal neoplasia.What influences the field effect? Diet?

32. Age related hypermethylation Inter-individual variation possibly due to diet?

35. What is going on in humans?Treatment of cells with polyphenols can reduce hypermethylatiion by inhibition of dnmt1

37. Impact of western diet highlighted by increased CRC incidence in Japan

38. Risk of CRC increased by obesity

41. Determine if faecal and tissue methylation correlate?

42. Are there significant differences in faecal methylation between patient groups?

43. Predicting disease status

46. Indicated that a panel of genes may be necessary

47. Majority of studies performed using non- or semi-quantitative assays

49. 19 Cancer patients

50. 18 Adenoma patients

51. 20 Patients with no-neoplasia

52. Faecal samples from 169 health individuals

53. ~10 males and 10 females/decade until 80 years old

55. Predicting disease status Multinomial modelling Age + ESR1 Overall = 77% Sensitivity = 63% , Specificity = 84% ESR1 and p14 Overall = 71% Sensitivity = 42%, Specificity = 86%

56. Correlation between tissue and faecal samples ESR1 was the only CGI to correlate

58. Unable to significantly predict disease status from faecal methylation profiles

60. Age difference between patient groups

61. Use of patients with no-neoplasia as controls

64. Determine if faecal and tissue methylation correlate?

65. Are there significant differences in faecal methylation between patient groups?

66. Predicting disease status

67. Could surrogate markers be used in dietary intervention studies?    ?

68. Acknowledgments IFR Nigel Belshaw Ian Johnson Liz Lund KasiaPrzybylska Jack Dainty Newcastle university John Mathers Amanda Coupe Wendy Bal Wansbeck Hospital D M Bradburn TNO Netherlands Robert Kleeman Funding FSA BBSRC CSG NuGO