PITX2 deficiency leads to atrial mitochondrial dysfunction

Cardiovasc Res. 2024 Dec 4;120(15):1907-1923. doi: 10.1093/cvr/cvae169.

Abstract

Aims: Reduced left atrial PITX2 is associated with atrial cardiomyopathy and atrial fibrillation (AF). PITX2 is restricted to left atrial cardiomyocytes (aCMs) in the adult heart. The links between PITX2 deficiency, atrial cardiomyopathy, and AF are not fully understood.

Methods and results: To identify mechanisms linking PITX2 deficiency to AF, we generated and characterized PITX2-deficient human aCMs derived from human induced pluripotent stem cells (hiPSC) and their controls. PITX2-deficient hiPSC-derived atrial cardiomyocytes showed shorter and disorganized sarcomeres and increased mononucleation. Electron microscopy found an increased number of smaller mitochondria compared with isogenic controls. Mitochondrial protein expression was altered in PITX2-deficient hiPSC-derived atrial cardiomyocytes. Single-nuclear RNA-sequencing found differences in cellular respiration pathways and differentially expressed mitochondrial and ion channel genes in PITX2-deficient hiPSC-derived atrial cardiomyocytes. PITX2 repression in hiPSC-derived atrial cardiomyocytes replicated dysregulation of cellular respiration. Mitochondrial respiration was shifted to increased glycolysis in PITX2-deficient hiPSC-derived atrial cardiomyocytes. PITX2-deficient human hiPSC-derived atrial cardiomyocytes showed higher spontaneous beating rates. Action potential duration was more variable with an overall prolongation of early repolarization, consistent with metabolic defects. Gene expression analyses confirmed changes in mitochondrial genes in left atria from 42 patients with AF compared with 43 patients with sinus rhythm. Dysregulation of left atrial mitochondrial (COX7C) and metabolic (FOXO1) genes was associated with PITX2 expression in human left atria.

Conclusion: PITX2 deficiency causes atrial mitochondrial dysfunction and a metabolic shift to glycolysis in human aCMs. PITX2-dependent metabolic changes can contribute to the structural and functional defects found in PITX2-deficient atria.

Keywords: PITX2; Atrial fibrillation; Human heart tissue; Human induced pluripotent stem cells; Metabolic shift; Mitochondrial dysfunction.

MeSH terms

  • Action Potentials
  • Atrial Fibrillation* / genetics
  • Atrial Fibrillation* / metabolism
  • Atrial Fibrillation* / pathology
  • Atrial Fibrillation* / physiopathology
  • Cardiomyopathies / genetics
  • Cardiomyopathies / metabolism
  • Cardiomyopathies / pathology
  • Cardiomyopathies / physiopathology
  • Case-Control Studies
  • Cell Respiration
  • Cells, Cultured
  • Energy Metabolism / genetics
  • Glycolysis
  • Heart Atria* / metabolism
  • Heart Atria* / pathology
  • Heart Atria* / physiopathology
  • Heart Rate
  • Homeobox Protein PITX2*
  • Homeodomain Proteins* / genetics
  • Homeodomain Proteins* / metabolism
  • Humans
  • Induced Pluripotent Stem Cells* / metabolism
  • Male
  • Mitochondria, Heart* / genetics
  • Mitochondria, Heart* / metabolism
  • Mitochondria, Heart* / pathology
  • Mitochondrial Proteins / deficiency
  • Mitochondrial Proteins / genetics
  • Mitochondrial Proteins / metabolism
  • Myocytes, Cardiac* / metabolism
  • Myocytes, Cardiac* / pathology
  • Transcription Factors* / genetics
  • Transcription Factors* / metabolism

Substances

  • Homeobox Protein PITX2
  • Transcription Factors
  • Homeodomain Proteins
  • Mitochondrial Proteins