Metabolomics and transcriptomics to identify pathways associated

Metabolomics and transcriptomics to identify
pathways associated with enhanced exercise
capacity in the rat
Charles Burant, MD, PhD
Department of Internal Medicine
Acknowledgments
Burant Lab
Katie Overmyer
Erin Shellman
Mary Treutelaar
Jinghua Xu
Sydney Bridges
Angela Subauste
Tanu Soni
Charles Evans
Steve Britton
Lauren Koch
Naisyin Wang
Funding
DK089503 (MNORC)
DK079084
DK077200
DA021519
U24 DK097153 (MRC)2
Robert C. and Veronica
Atkins Foundation
A. Alfred Taubman
Institute
((MRC)2
Metabolomics Core
Sub Pennathur
Steve Brown
Jaemin Byun
Jie “Jane” Cao
Arun Das
Chunhai Ruan
Sasha Raskind
Heidi Baum
Keri Bonds
Anu Janga
TM Rajendiran
Metscape
Alla Karnovsky
Barbara Mirel
Brian Athey
Terry Weymouth
Tim Hull
Glenn Tarcea
Alex Ade
Zach Wright
Maureen Sartor
Alex Terzian
Marci Brandenburg
Gilbert Omenn
HCR vs LCR: Bred to Run
HCR
LCR
Compared to LCR, HCR rats:
•
•
•
•
•
~ 4x running capacity
Eat the same, gain less weight
‘Healthier’ metabolic phenotypes
Live ~33% longer (but have no friends)
Utilize fatty acids for fuel when exercising
LCR
HCR
12 Month Caloric Restriction in HCR & LCR
LCR‐AL
600
550
Weekly Body Weight (g)
LCR‐CR
HCR‐AL
500
450
HCR‐CR
400
350
300
250
200
150
100
50
0
Weeks of Study
Untargeted metabolomics
70% cutoff
3998 features
85% cutoff
3534 features
100% cutoff
2432 features
Results of Untargeted Metabolomic Profiling in HCR/LCR
Metscape 2
A plugin for Cytoscape (http://cytoscape.org )
Uses EHMN metabolic reconstruction and KEGG
Gene Set Enrichment Analysis
GSEA,
LRpath
Metabolomics Data
Gene Expression Data
Compound Network
Compound ‐ Reaction
Network
Compound ‐ Reaction‐
Enzyme ‐ Gene Network
Karnovsky A, et al. 2012. Metscape 2 bioinformatics tool for the analysis and visualization of metabolomics and gene expression data. Bioinformatics. Feb 1;28(3):373‐80.
Compound ‐ Gene
Network
http://metscape.ncibi.org/
Metabolites, RNA Expression, Pathways
Summary
HCR rats have an increase in exercise capacity and select fatty acids for oxidation
Gene expression shows an increase in fatty acid oxidizing genes in skeletal muscle
Metabolomic measurements directly show increased fatty acid utilization while running
Gene expression also demonstrates increased BCAA metabolizing pathways
Metabolomic measurements show increased BCAA utilization during running
We hypothesize that BCAA transamination allows for greater expansion of span
2 TCA cycle intermediates during exercise and thereby supplies the oxalacetate to allow for fatty acid‐derived acetyl‐CoA to enter into the TCA cycle.
What is the upstream lesion?