1. Oncogene integration of “omic” networks to increase mitochondrial function, cell cycle entry and tumor progression. Fionnuala Morrish, Ph.D. Clinical Division Fred Hutchinson Cancer Research Center

2. Outline of talk Interconnection of –omic networks Interconnection of –omic networks The oncogene c-Myc The oncogene c-Myc Regulation of mitochondrial function and cell metabolism during cell cycle entry Regulation of mitochondrial function and cell metabolism during cell cycle entry Regulation of substrate supply for protein acetylation Regulation of substrate supply for protein acetylation Regulation of metabolism during tumor progression and regression Regulation of metabolism during tumor progression and regression

3. Interconnected –omic networks Transcriptome Metabolome Proteome Metabolite modified Proteome Palmitoylation Acetylation O-GlcNAcylation Farnsylation

4. The oncogene c-Myc Transcription factor regulating 15% of genome Transcription factor regulating 15% of genome Chromatin remodelling Chromatin remodelling Embryonic stem cell re-programming Embryonic stem cell re-programming Cell cycle regulator Cell cycle regulator Pervasive oncogene Pervasive oncogene Endogenous Myc required for tumor maintenance Endogenous Myc required for tumor maintenance

5. Myc regulation of mitochondrial function and cell metabolism during serum induced cell cycle entry c-myc +/+c-myc-/- 0hr 16hr0hr16hr Landay et al 2000

6. Mitochondrial genes targets induced by Myc on cell cycle entry Morrish et al. Genes and Development 2003

7. Network of genes regulated by Myc

8. Myc bioenergetic phenotype Myc+/+ Myc-/-MycER Myc-/- Myc-/-E1A Oxygen consumption and extracellular acidification rate

9. Bioenergetic cell cycle checkpoints

10. Experimental design for analysis of metabolism during cell cycle entry Myc-/-Myc+/+Myc-/-MycER Quiescent Cell cycle analysis ATP Pyruvate Lactate Glucose Oxygen Plate in 10% serum Mitochondrial mass And potential

11. 3 hr 8 hrs 16 hrs 3 hr 8 hrs 16 hrs Temporal regulation of cellular bioenergetics Morrish et al. Cell Cycle 2008 3 hr 8 hr16 hr Cell cycle entry Quiescent Serum addition

12. Mitochondrial function is required during Myc induced cell cycle entry 2-DG Oligomycin Rotenone

13. Myc regulates metabolic cell cycle checkpoints in response to serum Mitochondrial biogenesis and function Mitochondrial biogenesis and function Increased membrane potential Increased membrane potential Increased mitochondrial mass Increased mitochondrial mass Increased oxygen consumption Increased oxygen consumption Increased ATP Increased ATP Glycolysis Glycolysis Increased lactate production Increased lactate production Increased glucose uptake Increased glucose uptake

14. Myc regulation of substrate supply for protein acetylation

15. Myc regulation of gene transcription in interconnected metabolic networks Morrish et al. Cell Cycle 2008

16. Myc regulated flux increases acetyl CoA production from mitochondria

17. Myc regulation of histone acetylation TCA cycle [U- 13 C] Glucose [U- 13 C]-Pyruvate 13 C-FA, 13 C-acetate and 13 C-acetoacetate 13 C-Ac GCN5 [U- 13 C]-Acetyl-CoA [U- 13 C]-Citrate Nuclear histones