1. Companion PowerPoint slide set Obesity-associated breast cancer risk: a role for epigenetics? This teacher slide set was created by Dana Haine, MS, of the UNC Center for Environmental Health and Susceptibility, which is funded by the National Institute of Environmental Health Sciences (P30-ES010126).

2. Dr. Liza Makowski Studies Basal-like Breast Cancer (BBC) Aggressive cancer No specific drug therapies Short survival time if it spreads beyond breast tissue (metastasis) More prevalent in young pre-menopausal women More prevalent in African Americans Obesity is a risk factor for all breast cancers

3. Obesity causes BBC tumors to form at a faster rate compared to lean mice Number of weeks until tumor formation Fat-fed (Lean) Fat-fed (obese)

4. Cross-section of mammary (milk) duct http://www.womenshealth.gov/breast-cancer/what-is-breast-cancer/

5. Inside (lumen) of milk duct Label epithelial cells, fat cells and fibroblasts on the diagram below:

6. Epithelial Cells of Milk Duct Inside (lumen) of milk duct Fibroblasts Adipocytes (Fat cells) Immune cells are also present but are not shown in order to simplify schematic of the stroma. Stroma of mammary tissue

7. microenvironment

8. Cancer & the microenvironment The microenvironment (“soil”) surrounding a cell (“seed”) plays a role in tumor formation. HEALTHY CELL CANCER CELL “How does obesity alter the microenvironment in breast cancer?” Liza Makowski:

9. Cell to cell communication HGF-cMET paracrine signaling HGF C-met receptor Adapted from Melissa Troester/Patricia Casbas-Hernandez @ UNC

10. Cell to cell communication HGF-cMET paracrine signaling HGF cMET receptor Promotes Cell Division Promotes motility (leads to Metastasis) Promotes growth of blood vessels to supply tumor Prevents cell death Melissa Troester/Patricia Casbas-Hernandez @ UNC P HGF C-met

11. Sundaram, S., Makowski, L., et. al. Cancer Prevention Research (Under Review, 2012) Obesity increases levels of HGF mRNA and protein in normal mammary tissue *

12. Lean Mice Obese Mice Normal Cancer- associated Isolate Fibroblasts Culture Fibroblasts HGF Secretion Measure Ex vivo cell culture model Cell Division

13. HGF secretion in cultured cells

14. Lean Mice Obese Mice Normal Cancer- associated Isolate Fibroblasts Culture Fibroblasts HGF Secretion

15. Analysis of c-Met protein levels in BBC tumor cell line C. c-Met Control Phospho-c-Met BBC Tumor cell line BBC Tumor cell line + CM BBC Tumor cell line + mHGF

16. Fat cellFibroblast HGF Secretion c-Met activation Cancer formation “How does obesity alter the microenvironment in breast cancer?” Microenvironment Signal Epithelial cell Conclusion: growth factor signaling pathway (HGF/cMet) was upregulated with obesity.