Macrophage-mediated IL-6 signaling drives ryanodine receptor-2 calcium leak in postoperative atrial fibrillation

J Clin Invest. 2025 Mar 6;135(9):e187711. doi: 10.1172/JCI187711. eCollection 2025 May 1.

Abstract

Postoperative atrial fibrillation (poAF) is AF occurring days after surgery, with a prevalence of 33% among patients undergoing open-heart surgery. The degree of postoperative inflammation correlates with poAF risk, but less is known about the cellular and molecular mechanisms driving postoperative atrial arrhythmogenesis. We performed single-cell RNA-seq comparing atrial nonmyocytes from mice with and without poAF, which revealed infiltrating CCR2+ macrophages to be the most altered cell type. Pseudotime trajectory analyses identified Il-6 as a gene of interest driving in macrophages, which we confirmed in pericardial fluid collected from human patients after cardiac surgery. Indeed, macrophage depletion and macrophage-specific Il6ra conditional knockout (cKO) prevented poAF in mice. Downstream STAT3 inhibition with TTI-101 and cardiomyocyte-specific Stat3 cKO rescued poAF, indicating a proarrhythmogenic role of STAT3 in poAF development. Confocal imaging in isolated atrial cardiomyocytes (ACMs) uncovered what we believe to be a novel link between STAT3 and CaMKII-mediated ryanodine receptor-2 (RyR2)-Ser(S)2814 phosphorylation. Indeed, nonphosphorylatable RyR2S2814A mice were protected from poAF, and CaMKII inhibition prevented arrhythmogenic Ca2+ mishandling in ACMs from mice with poAF. Altogether, we provide multiomic, biochemical, and functional evidence from mice and humans that IL-6-STAT3-CaMKII signaling driven by infiltrating atrial macrophages is a pivotal driver of poAF, which portends therapeutic utility for poAF prevention.

Keywords: Arrhythmias; Calcium signaling; Cardiology; Immunology; Macrophages.

MeSH terms

  • Animals
  • Atrial Fibrillation* / etiology
  • Atrial Fibrillation* / genetics
  • Atrial Fibrillation* / metabolism
  • Atrial Fibrillation* / pathology
  • Calcium Signaling*
  • Calcium* / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / genetics
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / metabolism
  • Female
  • Humans
  • Interleukin-6* / genetics
  • Interleukin-6* / metabolism
  • Macrophages* / metabolism
  • Macrophages* / pathology
  • Male
  • Mice
  • Mice, Knockout
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / pathology
  • Postoperative Complications* / genetics
  • Postoperative Complications* / metabolism
  • Postoperative Complications* / pathology
  • Receptors, Interleukin-6 / genetics
  • Receptors, Interleukin-6 / metabolism
  • Ryanodine Receptor Calcium Release Channel* / genetics
  • Ryanodine Receptor Calcium Release Channel* / metabolism
  • STAT3 Transcription Factor / genetics
  • STAT3 Transcription Factor / metabolism
  • Signal Transduction*

Substances

  • Ryanodine Receptor Calcium Release Channel
  • Interleukin-6
  • STAT3 Transcription Factor
  • ryanodine receptor 2. mouse
  • Calcium
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Stat3 protein, mouse
  • RyR2 protein, human
  • interleukin-6, mouse
  • Il6ra protein, mouse
  • STAT3 protein, human
  • IL6 protein, human
  • Receptors, Interleukin-6