Isotopically nonstationary 13C flux analysis of Myc-induced metabolic reprogramming in B-cells

Taylor A. Murphy, Chi V. Dang, Jamey D. Young

Research output: Contribution to journalArticle

Abstract

We assessed several methods of 13C metabolic flux analysis (MFA) and found that isotopically nonstationary MFA achieved maximum flux resolution in cultured P493-6 B-cells, which have been engineered to provide tunable expression of the Myc oncoprotein. Comparison of metabolic flux maps obtained under oncogenic (High) and endogenous (Low) Myc expression levels revealed network-wide reprogramming in response to ectopic Myc expression. High Myc cells relied more heavily on mitochondrial oxidative metabolism than Low Myc cells and globally upregulated their consumption of amino acids relative to glucose. TCA cycle and amphibolic mitochondrial pathways exhibited 2- to 4-fold flux increases in High Myc cells, in contrast to modest increases in glucose uptake and lactate excretion. Because our MFA approach relied exclusively upon isotopic measurements of protein-bound amino acids and RNA-bound ribose, it is readily applicable to more complex tumor models that are not amenable to direct extraction and isotopic analysis of free intracellular metabolites.

Original languageEnglish (US)
Pages (from-to)206-217
Number of pages12
JournalMetabolic Engineering
Volume15
Issue number1
DOIs
StatePublished - Jan 2013
Externally publishedYes

Fingerprint

Metabolic Flux Analysis
B-Lymphocytes
Cells
Fluxes
Amino Acids
Glucose
Ribose
Oncogene Proteins
Amino acids
Lactic Acid
RNA
Metabolites
Metabolism
Tumors
Neoplasms
Proteins

Keywords

  • Cancer metabolism
  • Isotopomer analysis
  • Lymphoma
  • Mass spectrometry
  • Metabolic flux analysis
  • Warburg effect

ASJC Scopus subject areas

  • Bioengineering
  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Isotopically nonstationary 13C flux analysis of Myc-induced metabolic reprogramming in B-cells. / Murphy, Taylor A.; Dang, Chi V.; Young, Jamey D.

In: Metabolic Engineering, Vol. 15, No. 1, 01.2013, p. 206-217.

Research output: Contribution to journalArticle

Murphy, Taylor A. ; Dang, Chi V. ; Young, Jamey D. / Isotopically nonstationary 13C flux analysis of Myc-induced metabolic reprogramming in B-cells. In: Metabolic Engineering. 2013 ; Vol. 15, No. 1. pp. 206-217.
@article{48d9188f07204dc2ae93e2b8fbe93511,
title = "Isotopically nonstationary 13C flux analysis of Myc-induced metabolic reprogramming in B-cells",
abstract = "We assessed several methods of 13C metabolic flux analysis (MFA) and found that isotopically nonstationary MFA achieved maximum flux resolution in cultured P493-6 B-cells, which have been engineered to provide tunable expression of the Myc oncoprotein. Comparison of metabolic flux maps obtained under oncogenic (High) and endogenous (Low) Myc expression levels revealed network-wide reprogramming in response to ectopic Myc expression. High Myc cells relied more heavily on mitochondrial oxidative metabolism than Low Myc cells and globally upregulated their consumption of amino acids relative to glucose. TCA cycle and amphibolic mitochondrial pathways exhibited 2- to 4-fold flux increases in High Myc cells, in contrast to modest increases in glucose uptake and lactate excretion. Because our MFA approach relied exclusively upon isotopic measurements of protein-bound amino acids and RNA-bound ribose, it is readily applicable to more complex tumor models that are not amenable to direct extraction and isotopic analysis of free intracellular metabolites.",
keywords = "Cancer metabolism, Isotopomer analysis, Lymphoma, Mass spectrometry, Metabolic flux analysis, Warburg effect",
author = "Murphy, {Taylor A.} and Dang, {Chi V.} and Young, {Jamey D.}",
year = "2013",
month = "1",
doi = "10.1016/j.ymben.2012.07.008",
language = "English (US)",
volume = "15",
pages = "206--217",
journal = "Metabolic Engineering",
issn = "1096-7176",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Isotopically nonstationary 13C flux analysis of Myc-induced metabolic reprogramming in B-cells

AU - Murphy, Taylor A.

AU - Dang, Chi V.

AU - Young, Jamey D.

PY - 2013/1

Y1 - 2013/1

N2 - We assessed several methods of 13C metabolic flux analysis (MFA) and found that isotopically nonstationary MFA achieved maximum flux resolution in cultured P493-6 B-cells, which have been engineered to provide tunable expression of the Myc oncoprotein. Comparison of metabolic flux maps obtained under oncogenic (High) and endogenous (Low) Myc expression levels revealed network-wide reprogramming in response to ectopic Myc expression. High Myc cells relied more heavily on mitochondrial oxidative metabolism than Low Myc cells and globally upregulated their consumption of amino acids relative to glucose. TCA cycle and amphibolic mitochondrial pathways exhibited 2- to 4-fold flux increases in High Myc cells, in contrast to modest increases in glucose uptake and lactate excretion. Because our MFA approach relied exclusively upon isotopic measurements of protein-bound amino acids and RNA-bound ribose, it is readily applicable to more complex tumor models that are not amenable to direct extraction and isotopic analysis of free intracellular metabolites.

AB - We assessed several methods of 13C metabolic flux analysis (MFA) and found that isotopically nonstationary MFA achieved maximum flux resolution in cultured P493-6 B-cells, which have been engineered to provide tunable expression of the Myc oncoprotein. Comparison of metabolic flux maps obtained under oncogenic (High) and endogenous (Low) Myc expression levels revealed network-wide reprogramming in response to ectopic Myc expression. High Myc cells relied more heavily on mitochondrial oxidative metabolism than Low Myc cells and globally upregulated their consumption of amino acids relative to glucose. TCA cycle and amphibolic mitochondrial pathways exhibited 2- to 4-fold flux increases in High Myc cells, in contrast to modest increases in glucose uptake and lactate excretion. Because our MFA approach relied exclusively upon isotopic measurements of protein-bound amino acids and RNA-bound ribose, it is readily applicable to more complex tumor models that are not amenable to direct extraction and isotopic analysis of free intracellular metabolites.

KW - Cancer metabolism

KW - Isotopomer analysis

KW - Lymphoma

KW - Mass spectrometry

KW - Metabolic flux analysis

KW - Warburg effect

UR - http://www.scopus.com/inward/record.url?scp=84872376676&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84872376676&partnerID=8YFLogxK

U2 - 10.1016/j.ymben.2012.07.008

DO - 10.1016/j.ymben.2012.07.008

M3 - Article

C2 - 22898717

AN - SCOPUS:84872376676

VL - 15

SP - 206

EP - 217

JO - Metabolic Engineering

JF - Metabolic Engineering

SN - 1096-7176

IS - 1

ER -