PDE5 inhibition rescues mitochondrial dysfunction and angiogenic responses induced by Akt3 inhibition by promotion of PRC expression

J Biol Chem. 2020 Dec 25;295(52):18091-18104. doi: 10.1074/jbc.RA120.013716. Epub 2020 Oct 21.

Abstract

Akt3 regulates mitochondrial content in endothelial cells through the inhibition of PGC-1α nuclear localization and is also required for angiogenesis. However, whether there is a direct link between mitochondrial function and angiogenesis is unknown. Here we show that Akt3 depletion in primary endothelial cells results in decreased uncoupled oxygen consumption, increased fission, decreased membrane potential, and increased expression of the mitochondria-specific protein chaperones, HSP60 and HSP10, suggesting that Akt3 is required for mitochondrial homeostasis. Direct inhibition of mitochondrial homeostasis by the model oxidant paraquat results in decreased angiogenesis, showing a direct link between angiogenesis and mitochondrial function. Next, in exploring functional links to PGC-1α, the master regulator of mitochondrial biogenesis, we searched for compounds that induce this process. We found that, sildenafil, a phosphodiesterase 5 inhibitor, induced mitochondrial biogenesis as measured by increased uncoupled oxygen consumption, mitochondrial DNA content, and voltage-dependent anion channel protein expression. Sildenafil rescued the effects on mitochondria by Akt3 depletion or pharmacological inhibition and promoted angiogenesis, further supporting that mitochondrial homeostasis is required for angiogenesis. Sildenafil also induces the expression of PGC-1 family member PRC and can compensate for PGC-1α activity during mitochondrial stress by an Akt3-independent mechanism. The induction of PRC by sildenafil depends upon cAMP and the transcription factor CREB. Thus, PRC can functionally substitute during Akt3 depletion for absent PGC-1α activity to restore mitochondrial homeostasis and promote angiogenesis. These findings show that mitochondrial homeostasis as controlled by the PGC family of transcriptional activators is required for angiogenic responses.

Keywords: Akt PKB; Akt3; Akt3/PKBγ; PGC-1α; PPARGC1A; PRC; angiogenesis; endothelial cells; mitochondria; peroxisome proliferator–activated receptor γ coactivator-1 α; phosphodiesterases.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / drug effects
  • Cyclic Nucleotide Phosphodiesterases, Type 5 / chemistry*
  • Endothelium, Vascular / drug effects*
  • Gene Expression Regulation
  • Humans
  • Mitochondria / drug effects*
  • Mitochondria / pathology
  • Neovascularization, Physiologic / drug effects*
  • Organelle Biogenesis
  • Oxygen Consumption
  • Phosphodiesterase 5 Inhibitors / pharmacology*
  • Proto-Oncogene Proteins c-akt / antagonists & inhibitors*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • Phosphodiesterase 5 Inhibitors
  • Transcription Factors
  • peroxisome-proliferator-activated receptor-gamma coactivator-1
  • AKT3 protein, human
  • Proto-Oncogene Proteins c-akt
  • Cyclic Nucleotide Phosphodiesterases, Type 5
  • PDE5A protein, human