Oncogene integration of Oncogene integration of “omic” networks to increase “omic” networks to increase mitochondrial function, cell mitochondrial function, cell

cycle entry and tumor cycle entry and tumor progression.progression.

Fionnuala Morrish, Ph.D.Fionnuala Morrish, Ph.D.Clinical DivisionClinical Division

Fred Hutchinson Cancer Research Fred Hutchinson Cancer Research CenterCenter

Outline of talkOutline of talk

Interconnection of –omic networksInterconnection of –omic networks The oncogene c-MycThe oncogene c-Myc Regulation of mitochondrial function Regulation of mitochondrial function

and cell metabolism during cell cycle and cell metabolism during cell cycle entryentry

Regulation of substrate supply for Regulation of substrate supply for protein acetylationprotein acetylation

Regulation of metabolism during Regulation of metabolism during tumor progression and regressiontumor progression and regression

Interconnected –omic Interconnected –omic networksnetworks

Transcriptome Metabolome

Proteome

Metabolite modified Proteome

PalmitoylationAcetylationO-GlcNAcylationFarnsylation

The oncogene c-MycThe oncogene c-Myc

Transcription factor regulating 15% Transcription factor regulating 15% of genomeof genome

Chromatin remodellingChromatin remodelling Embryonic stem cell re-programmingEmbryonic stem cell re-programming Cell cycle regulatorCell cycle regulator Pervasive oncogenePervasive oncogene Endogenous Myc required for tumor Endogenous Myc required for tumor

maintenance maintenance

Myc regulation of mitochondrial Myc regulation of mitochondrial function and cell metabolism function and cell metabolism

during serum induced cell cycle during serum induced cell cycle entryentry

c-myc +/+ c-myc-/-

0hr 16hr 0hr 16hr

Landay et al 2000

Mitochondrial genes targets Mitochondrial genes targets induced by Myc on cell cycle induced by Myc on cell cycle

entryentry

Morrish et al. Genes and Development 2003

Network of genes regulated by Network of genes regulated by MycMyc

Myc bioenergetic phenotypeMyc bioenergetic phenotype

Myc+/+

Myc-/-MycER

Myc-/-

Myc-/-E1A

Oxygen consumptionand extracellular acidification rate

Bioenergetic cell cycle Bioenergetic cell cycle checkpointscheckpoints

Experimental design for Experimental design for analysis of metabolism during analysis of metabolism during

cell cycle entrycell cycle entry

Myc-/- Myc+/+ Myc-/-MycER

Quiescent

Cell cycle analysis

ATPPyruvate

Lactate

GlucoseOxygen

Plate in 10% serum

Mitochondrial massAnd potential

3 hr8 hrs16 hrs

3 hr8 hrs16 hrs

Temporal regulation of cellular bioenergetics

Morrish et al. Cell Cycle 2008

3 hr 8 hr 16 hr

Cell cycle entryQuiescent

Serum addition

Mitochondrial function is Mitochondrial function is required during Myc induced required during Myc induced

cell cycle entrycell cycle entry

2-DG

Oligomycin

Rotenone

Myc regulates metabolic cell Myc regulates metabolic cell cycle checkpoints in response cycle checkpoints in response

to serumto serum Mitochondrial biogenesis and functionMitochondrial biogenesis and function

Increased membrane potentialIncreased membrane potential Increased mitochondrial massIncreased mitochondrial mass Increased oxygen consumptionIncreased oxygen consumption Increased ATPIncreased ATP

GlycolysisGlycolysis Increased lactate productionIncreased lactate production Increased glucose uptakeIncreased glucose uptake

Myc regulation of Myc regulation of substrate supply for substrate supply for protein acetylationprotein acetylation

Myc regulation of gene transcription in Myc regulation of gene transcription in interconnected metabolic networksinterconnected metabolic networks

Morrish et al. Cell Cycle 2008

Myc regulated flux increases Myc regulated flux increases acetyl CoA production from acetyl CoA production from

mitochondria mitochondria

Myc regulation of histone Myc regulation of histone acetylationacetylation

TCA cycle

[U-13C] Glucose

[U-13C]-Pyruvate 13C-FA, 13C-acetate and 13C-acetoacetate

13C-Ac13C-Ac

GCN5

[U-13C]-Acetyl-CoA

[U-13C]-Citrate

Nuclear histones