Phasic metabolic remodeling coordinates cardiac myofibroblast differentiation and function.
4 affiliations
- 1Center for Cardiometabolic Science, Christina Lee Brown Envirome Institute, University of Louisville School of Medicine, Louisville, KY, USA; Department of Biochemistry and Molecular Genetics, University of Louisville School of Medicine, Louisville, KY, USA.
- 2Center for Cardiometabolic Science, Christina Lee Brown Envirome Institute, University of Louisville School of Medicine, Louisville, KY, USA; Department of Physiology, University of Louisville, Louisville, KY, USA.
- 3Center for Cardiometabolic Science, Christina Lee Brown Envirome Institute, University of Louisville School of Medicine, Louisville, KY, USA.
- 4Center for Cardiometabolic Science, Christina Lee Brown Envirome Institute, University of Louisville School of Medicine, Louisville, KY, USA. Electronic address: [email protected].
Abstract
Fibroblast differentiation into matrix-producing, contractile myofibroblasts is accompanied by metabolic reprogramming, but whether these changes precede differentiation or arise from the activated phenotype remains unclear. We integrated time-resolved extracellular flux analysis, transcriptomics, stable isotope-resolved metabolomics, and metabolic interventions to define the relationship between metabolism and myofibroblast differentiation in adult mouse cardiac fibroblasts stimulated with transforming growth factor-β (TGFβ). TGFβ increased glycolysis within 1.5 h, before broad transcriptional remodeling and induction of periostin and α-smooth muscle actin. Mitochondrial respiration initially declined but increased at 48 h as the phenotype emerged, accompanied by smaller, less elongated mitochondria and greater respiratory capacity. [U-¹³C₆]-glucose tracing revealed progressive incorporation of glucose-derived carbon into glycolytic intermediates, nucleotide sugars, and glycerol 3-phosphate, followed later by increased labeling of tricarboxylic acid cycle intermediates. TGFβ-treated fibroblasts also demonstrated enhanced insulin-stimulated glycolysis. Replacing glucose with galactose attenuated collagen and periostin induction, whereas glyceraldehyde 3-phosphate dehydrogenase inhibition prevented induction of collagen, periostin, and α-smooth muscle actin. Conversely, glucose transporter 1 or mutant PFK2 (GlycoHi) overexpression increased glycolysis without inducing myofibroblast markers. Together, these loss- and gain-of-function studies indicate that glycolytic activity is required for complete differentiation but is not sufficient to initiate the phenotype. Inhibition of mitochondrial Complex I or ATP synthase preferentially reduced collagen abundance, while glycolytic and Complex I inhibition diminished TGFβ-induced collagen gel contraction. These findings define a phasic metabolic program in which early glycolytic activation precedes transcriptional and phenotypic remodeling, whereas mitochondrial respiratory capacity increases later and distinct metabolic nodes selectively regulate collagen abundance and contractile behavior.
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BibTeX
@article{wells2026phasic,
title = {{Phasic metabolic remodeling coordinates cardiac myofibroblast differentiation and function.}},
author = {Collin K Wells and Lianay Gutierrez Luque and Daniel C Nguyen and Madison S Teer and Ning Chen and Yania Martinez-Ondaro and Pawel K Lorkiewicz and Richa Singhal and Maleesha De Silva and Bradford G Hill},
journal = {Redox biology},
year = {2026},
month = oct,
doi = {10.1016/j.redox.2026.104441},
url = {https://europepmc.org/article/MED/42893628},
note = {CC BY},
}
RIS
TY - JOUR
TI - Phasic metabolic remodeling coordinates cardiac myofibroblast differentiation and function.
AU - Collin K Wells
AU - Lianay Gutierrez Luque
AU - Daniel C Nguyen
AU - Madison S Teer
AU - Ning Chen
AU - Yania Martinez-Ondaro
AU - Pawel K Lorkiewicz
AU - Richa Singhal
AU - Maleesha De Silva
AU - Bradford G Hill
PY - 2026
DA - 2026/10/16
JO - Redox biology
DO - 10.1016/j.redox.2026.104441
UR - https://europepmc.org/article/MED/42893628
AB - Fibroblast differentiation into matrix-producing, contractile myofibroblasts is accompanied by metabolic reprogramming, but whether these changes precede differentiation or arise from the activated phenotype remains unclear. We integrated time-resolved extracellular flux analysis, transcriptomics, stable isotope-resolved metabolomics, and metabolic interventions to define the relationship between metabolism and myofibroblast differentiation in adult mouse cardiac fibroblasts stimulated with transforming growth factor-β (TGFβ). TGFβ increased glycolysis within 1.5 h, before broad transcriptional remodeling and induction of periostin and α-smooth muscle actin. Mitochondrial respiration initially declined but increased at 48 h as the phenotype emerged, accompanied by smaller, less elongated mitochondria and greater respiratory capacity. [U-¹³C₆]-glucose tracing revealed progressive incorporation of glucose-derived carbon into glycolytic intermediates, nucleotide sugars, and glycerol 3-phosphate, followed later by increased labeling of tricarboxylic acid cycle intermediates. TGFβ-treated fibroblasts also demonstrated enhanced insulin-stimulated glycolysis. Replacing glucose with galactose attenuated collagen and periostin induction, whereas glyceraldehyde 3-phosphate dehydrogenase inhibition prevented induction of collagen, periostin, and α-smooth muscle actin. Conversely, glucose transporter 1 or mutant PFK2 (GlycoHi) overexpression increased glycolysis without inducing myofibroblast markers. Together, these loss- and gain-of-function studies indicate that glycolytic activity is required for complete differentiation but is not sufficient to initiate the phenotype. Inhibition of mitochondrial Complex I or ATP synthase preferentially reduced collagen abundance, while glycolytic and Complex I inhibition diminished TGFβ-induced collagen gel contraction. These findings define a phasic metabolic program in which early glycolytic activation precedes transcriptional and phenotypic remodeling, whereas mitochondrial respiratory capacity increases later and distinct metabolic nodes selectively regulate collagen abundance and contractile behavior.
SN - 2213-2317
ER -