Integrative proteomics and phosphoproteomics in pulmonary arterial hypertension

Sci Rep. 2019 Dec 9;9(1):18623. doi: 10.1038/s41598-019-55053-6.

Abstract

Pulmonary arterial endothelial cells (PAEC) are mechanistically linked to origins of pulmonary arterial hypertension (PAH). Here, global proteomics and phosphoproteomics of PAEC from PAH (n = 4) and healthy lungs (n = 5) were performed using LC-MS/MS to confirm known pathways and identify new areas of investigation in PAH. Among PAH and control cells, 170 proteins and 240 phosphopeptides were differentially expressed; of these, 45 proteins and 18 phosphopeptides were located in the mitochondria. Pathologic pathways were identified with integrative bioinformatics and human protein-protein interactome network analyses, then confirmed with targeted proteomics in PAH PAEC and non-targeted metabolomics and targeted high-performance liquid chromatography of metabolites in plasma from PAH patients (n = 30) and healthy controls (n = 12). Dysregulated pathways in PAH include accelerated one carbon metabolism, abnormal tricarboxylic acid (TCA) cycle flux and glutamate metabolism, dysfunctional arginine and nitric oxide pathways, and increased oxidative stress. Functional studies in cells confirmed abnormalities in glucose metabolism, mitochondrial oxygen consumption, and production of reactive oxygen species in PAH. Altogether, the findings indicate that PAH is typified by changes in metabolic pathways that are primarily found in mitochondria.

Publication types

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

MeSH terms

  • Adult
  • Arginine / metabolism
  • Citric Acid Cycle
  • Computational Biology
  • Endothelial Cells / metabolism
  • Female
  • Glucose / metabolism
  • Humans
  • Lung / metabolism
  • Lung Transplantation
  • Male
  • Metabolomics
  • Middle Aged
  • Mitochondria / metabolism
  • Nitric Oxide / metabolism
  • Oxidative Stress
  • Peptides / metabolism*
  • Phosphoproteins / metabolism*
  • Protein Interaction Mapping
  • Proteome
  • Proteomics / methods*
  • Pulmonary Arterial Hypertension / metabolism*
  • Reactive Oxygen Species / metabolism

Substances

  • Peptides
  • Phosphoproteins
  • Proteome
  • Reactive Oxygen Species
  • Nitric Oxide
  • Arginine
  • Glucose