1. Chung S. Park Animal Science Department North Dakota State University November, 2012 Canola Oil and Breast Cancer Research Projects

2. Topics 1.Maternal lipotropes (methyl nutrients) diet and mammary cancer-Chung Park 2.Maternal canola oil supplementation and mammary cancer-Lawrence Mabasa 3.Update of Lipocan project progress-Lawrence Mabasa

3. Maternal lipotrope (methyl) and mammary carcinogenesis in offspring LIPOCAN= Lipotrope + Canola Oil

4. Our previous work Over the past 22 years, our laboratory has studied link between lipotrope and mammary cancer Lipotropes can alter the expression of genes that are associated with cell growth and metabolism in the mammary glands Manipulation of dietary lipotropes can affect susceptibility of virgin female rats to mammary cancer (Park et al., 2010. Advances in Medicine and Biology, Vol.12)

5. Lipotropes (methyl nutrients) Lipotropes are methyl (-CH 3 ) group containing amino acid and vitamins (methionine, choline, folate, and vitamin B 12 ) Play major role in one-carbon metabolism (DNA methylation and synthesis), which is involved in epigenetic imprinting

6. Epigenetics: imprint Permanent alterations in gene expression and metabolism that occur during early fetal development Epigenetic modifications are inheritable and affected by environment, notably nutrition/diet

7. Things written in pen you cant change. Thats DNA. Things written in pencil you can. Thats epigenetics.-Danielle Reed

8. Maternal nutrition & offspring health Maternal nutrition may lead to metabolic imprinting (biological memory) which can be passed on to offspring Prenatal and early postnatal nutrition Metabolic imprinting (program) Lifetime susceptibility to chronic disease in offspring

9. Maternal nutrition and breast cancer More complete and full mammary development during full-term pregnancy is closely associated with a long-term reduction of mammary cancer Maternal diet/nutrition Epigenetic modifications (moms, offspring) Mammary gland development Breast cancer risk

10. You are what your mother ate Your mothers diet during pregnancy can cause critical changes that stick with you into adulthood

11. Methyl nutrients and cancer studies Most studies have been concerned with the effects of methyl deprivation upon carcinogenesis However, dietary methyl deficiency has minimum relevancy because normal human diets are unlikely to be deficient

12. Methyl nutrients and cancer studies Studies address the effect of individual methyl nutrients (e.g., folic acid) on breast cancer However, there have been no studies which address the interplay among all four methyl nutrients on reduction of breast cancer Supplementation – novel concept

13. Maternal diet with 3x methyl nutrients Yellow mouse Reduced lifespan High risk of obesity, diabetes, and cancer Prolonged lifespan Lower risk of obesity, diabetes, and cancer Agouti mouse Cooney et al. (2002) J Nutr. Mom Offspring

14. Hypothesis In Utero exposure to lipotropes decreases mammary carcinogenesis in offspring

15. Funded by Department of Defense-Medical Research Program Findings

16. Mammary carcinogenesis in offspring ItemControlMethyl-diet Tumor incidence (%)10053.3* (47% ) Latency period (weeks)7.93 ± 0.3211.27 ± 0.85* (42% ) Tumor multiplicity (# of tumors/rat) 2.93 ± 0.21 1.20 ± 0.23* (59% ) (* P<0.05)

17. Summary In utero exposure to lipotrope supplementation: Significantly reduced chemically-induced mammary carcinogenesis in offspring The first experimental evidence linking maternal lipotropic diet (all four methyl nutrients) to reduced mammary carcinogenesis in offspring Published in Carcinogenesis June 2012

18. Lipotropes and mammary carcinogenesis Carcinogenesis 33:1106-1112 (2012) Research in Veterinary Science 93:259-263 (2012) In Advances in Medicine and Biology Vol.12:133-139 (2010) In Vitro Cell Dev Biol-Anim 44:268-272 (2008) Recent publications

19. Current and future studies ? IN Prenatal exposure to lipotropes Reduction of mammary carcinogenesis in offspring OUT Black box Currently funded by National Institutes of Health (2012-14)

20. Support Lipotrope and mammary cancer NIH-National Cancer Institute 1998, 2002, 2012 (active) DOD-Breast Cancer Research Program 1994, 2009 American Institute for Cancer Research 2003

21. Maternal canola oil supplementation and mammary cancer risk in offspring

22. Objective The objective of the study was to investigate the role of maternal dietary canola oil on the susceptibility of female offspring to mammary carcinogenesis

23. Protocol Pregnant rats were fed either of the following diets: Control diet (containing 10% soybean oil) Canola oil diet (10% canola oil) After weaning, offspring were fed the control diet and then injected with NMU (which is a chemical carcinogen) to induce mammary cancer

24. Results Mammary carcinogenesis data in offspring ItemControlCanolaP Tumor incidence (%)8773 ( 16%) Latency period (weeks) 11.08±0.9011.90±0.96 0.71 Tumor multiplicity/no. 1.93±0.32 1.02±0.250.04

25. Results * * * * *

26. Summary Maternal canola oil diet reduced overall mammary carcinogenesis in offspring Manuscript of this work is currently undergoing revision in the Journal of Nutrition

27. Lipotropes plus canola oil (lipocan) projects

28. Rationale Lipotropes Canola oil Supplementation Anti-breast cancer effect + Synergistic

29. Background In 2011, we received support from the NCGA to study the effect of lipocan (lipotropes plus canola oil) on mammary cancer risk

30. Strategy To complete the US patent application: Phase I: data gathering on the anticancer effect of lipocan Phase II: studying the mechanism, how lipocan reduces mammary cancer risk

31. Progress Currently two independent projects are ongoing: Experiment 1 deals with the effect of lipocan on virgin rats Experiment 2 involves the role of maternal lipocan on the risk of offspring to cancer

32. Time line Physical evidence, and data collection Mechanism and patent application Animal trials (experiment 1&2) Some tissue sample assays 2011 2012 2013 2014 PHASE I PHASE II Animal trials (experiment 2) Tissue sample assays Writing up reports

33. Implications Results may be useful in development of a nutraceutical product (lipocan) to reduce long term breast cancer risk in humans US patent royalties 30% Inventor: NCGA 30-50% NDSU Animal Sciences Department 20-35% NDSU Research Foundation

34. Support and Personnel Northern Canola Growers Association L. Mabasa K. Cho S. Bae M. Walters Dr. Park Acknowledgment