Chung S. ParkAnimal Science Department

North Dakota State UniversityNovember, 2012

Canola Oil and Breast Cancer Research Projects

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

Maternal lipotrope (methyl) and mammary carcinogenesis in offspring

LIPOCAN= Lipotrope + Canola Oil

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)

Lipotropes (methyl nutrients)

Lipotropes are methyl (-CH3) group containing amino acid and vitamins (methionine, choline, folate, and vitamin B12)

Play major role in one-carbon metabolism (DNA methylation and synthesis), which is involved in epigenetic imprinting

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

“Things written in pen you can’t change. That’s DNA. Things written in pencil you can. That’s epigenetics.”-Danielle Reed

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

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

“You are what your mother ate”

Your mother’s diet during pregnancy can cause critical changes

that stick with you into adulthood

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

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

Maternal diet with3x 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

Hypothesis

“In Utero exposure to lipotropes decreases mammary carcinogenesis in offspring”

Funded by Department of Defense-Medical Research Program

Findings

Mammary carcinogenesis in offspring

Item Control Methyl-diet

Tumor incidence (%) 100 53.3* (47% ↓)

Latency period (weeks) 7.93 ± 0.32 11.27 ± 0.85* (42% ↑)

Tumor multiplicity (# of tumors/rat) 2.93 ± 0.21 1.20 ± 0.23* (59% ↓)

(* P<0.05)

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

“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

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)

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

Maternal canola oil supplementation and mammary

cancer risk in offspring

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

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

Results

Mammary carcinogenesis data in offspring

Item Control Canola P

Tumor incidence (%) 8773

( 16%)

Latency period

(weeks)11.08±0.90

11.90±0.9

60.71

Tumor multiplicity/no. 1.93±0.32

1.02±0.250.04

Results

0100020003000400050006000700080009000

Control

Weeks after NMU injection

Tu

mo

r vo

lum

e (

mm

3)

* * * * *

Summary

Maternal canola oil diet reduced overall mammary carcinogenesis in offspring

Manuscript of this work is currently undergoing revision in the Journal of Nutrition

Lipotropes plus canola oil (lipocan) projects

Rationale

Lipotropes

Canola oil

Supplementation

Anti-breast cancer effect

+Synergistic

Background

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

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

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

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

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

Support and Personnel

Northern Canola Growers Association

L. Mabasa K. Cho S. Bae M. Walters Dr. Park

Acknowledgment