1. Influence of initial differentiated state of the normal cell on the final tumorigenic phenotype Yesenia Correa Undergraduate Research Program Cold Spring Harbor Laboratory Dr. Maia Chanrion Dr. Scott Powers

2. Influence of normal cell on neoplastic phenotype Normal Breast Tissue WIT medium MEGM medium BPEC Luminal HMEC Myoepithelial Transformation hTERT SV-40 LT/st H-RasV12 BPLER HMLERWeakly tumorigenic Non metastatic 10 4 fold more tumorigenic Lung metastases Ince et al., Cancer cell, 2007

3. Importance of normal cell state in the ability of Myc/p53- combination to transform hepatoblasts into malignancy Zender et al., CSH symposia on Quantitative Biology, 2005 P53-/- Tumor No Tumor Myc Day 14 Day 18 What change in the 4 days causes such a difference?

4. Phenotype Comparison D18/D18 Powers and Chanrion (Unpublished) D13/D13

5. Phenotype Comparison D18/D13 Powers and Chanrion (Unpublished)

6. Method of Ranking Pathways Scoring Metric Microarray Data Annotations A score for each Pathway: indicates how much it was affected by the condition Many Different Scoring Metrics Available

7. PPARA RXRA PPARG Adipocyte differentiation Adaptive thermogenesis Cell survival Ubiquitination Gluconeogenesis Lipid metabolism (ketogenesis, lipid transport, lipogenesis, cholesterol metabolism, fatty acid oxidation PPARG Pathway

8. Knockdown by shRNA No Tumor LMS-Myc D18 P53-/- Tumor sh PPAR pathway P53-/- ?

9. Activation of PPARG D13 Tumor LMS-Myc P53-/- No Tumor PPAR pathway Activation P53-/- ?

10. Shuttle shRNAs from V2 library into LMP vector 16 total LMP-vector shRNA 8 targeting PPARG 2 targeting PPARA 6 targeting RXRA Puro r P PGK GFPIRES LTR ++ X S R 5’miR303’miR30 B Unique sites: B = Bgl II R = EcoR I X = Xho I Bs = BstX I S = Sal I Bs PG6 100bp ladder PG8PG7 Ongoing experiments...

11. Potential cancer therapy: Activation of the PPARG Pathway using Troglitazone Explored as a potential cancer therapeutic in animal models and clinical trials Some but not all studies implicate its potential therapeutic utility in liver cancer Yu et al., Hepatology, 2006

12. Clonogenic Assay with Troglitazone Mouse Cell lines: –Day 14: 18-8/1 and 17-8/3 Tumor cell line from D14 p53-/- and myc hepatoblasts –Day 18: IM+DLCs, 122-2ve, and 118-1 Tumor cell line from D18 p53-/-, myc and shRNA hepatoblasts

13. Ongoing experiments Validation of PPARA, PPARG, and RXRA shRNA through Western Blot Biological validation (tumorigenicity assays) Determinant of troglitazone response?

14. Acknowledgements Dr. Scott PowersDr. Maia Chanrion C. Bliss Memorial Fund Cold Spring Harbor Laboratory Woodbury Genome Center Powers Lab Howard Hughes Medical Institute