TNFα increases the degradation of pyruvate dehydrogenase kinase 4 by the Lon protease to support proinflammatory genes

Nabil E. Boutagy, Joseph W. Fowler, Kariona A. Grabinska, Rebecca Cardone, Qiushi Sun, Kyla R. Vazquez, Michael B. Whalen, Xiaolong Zhu, Raja Chakraborty, Kathleen A. Martin, Michael Simons, Casey E. Romanoski, Richard G. Kibbey, William C. Sessa

Research output: Contribution to journalArticlepeer-review

Abstract

The endothelium is a major target of the proinflammatory cytokine, tumor necrosis factor alpha (TNFα). Exposure of endothelial cells (EC) to proinflammatory stimuli leads to an increase in mitochondrial metabolism; however, the function and regulation of elevated mitochondrial metabolism in EC in response to proinflammatory cytokines remain unclear. Studies using high-resolution metabolomics and 13C-glucose and 13C-glutamine labeling flux techniques showed that pyruvate dehydrogenase activity (PDH) and oxidative tricarboxylic acid cycle (TCA) flux are elevated in human umbilical vein ECs in response to overnight (16 h) treatment with TNFα (10 ng/mL). Mechanistic studies indicated that TNFα mediated these metabolic changes via mitochondrial-specific protein degradation of pyruvate dehydrogenase kinase 4 (PDK4, inhibitor of PDH) by the Lon protease via an NF-κB-dependent mechanism. Using RNA sequencing following siRNA-mediated knockdown of the catalytically active subunit of PDH, PDHE1α (PDHA1 gene), we show that PDH flux controls the transcription of approximately one-third of the genes that are up-regulated by TNFα stimulation. Notably, TNFα-induced PDH flux regulates a unique signature of proinflammatory mediators (cytokines and chemokines) but not inducible adhesion molecules. Metabolomics and ChIP sequencing for acetylated modification on lysine 27 of histone 3 (H3K27ac) showed that TNFα-induced PDH flux promotes histone acetylation of specific gene loci via citrate accumulation and ATP-citrate lyase–mediated generation of acetyl CoA. Together, these results uncover a mechanism by which TNFα signaling increases oxidative TCA flux of glucose to support TNFα-induced gene transcription through extramitochondrial acetyl CoA generation and histone acetylation.

Original languageEnglish (US)
Article numbere2218150120
JournalProceedings of the National Academy of Sciences of the United States of America
Volume120
Issue number38
DOIs
StatePublished - Sep 19 2023
Externally publishedYes

Keywords

  • TNF
  • endothelium
  • gene expression
  • inflammation
  • vascular

ASJC Scopus subject areas

  • General

Fingerprint

Dive into the research topics of 'TNFα increases the degradation of pyruvate dehydrogenase kinase 4 by the Lon protease to support proinflammatory genes'. Together they form a unique fingerprint.

Cite this