Targeting myeloid cell coagulation signaling blocks MAP kinase/TGF-β1–driven fibrotic remodeling in ischemic heart failure
Venkata Garlapati, Michael Molitor, Thomas Michna, Gregory S. Harms, Stefanie Finger, Rebecca Jung, Jeremy Lagrange, Panagiotis Efentakis, Johannes Wild, Maike Knorr, Susanne Karbach, Sabine Wild, Ksenija Vujacic-Mirski, Thomas Münzel, Andreas Daiber, Moritz Brandt, Tommaso Gori, Hendrik Milting, Stefan Tenzer, Wolfram Ruf, Philip Wenzel
Venkata Garlapati, Michael Molitor, Thomas Michna, Gregory S. Harms, Stefanie Finger, Rebecca Jung, Jeremy Lagrange, Panagiotis Efentakis, Johannes Wild, Maike Knorr, Susanne Karbach, Sabine Wild, Ksenija Vujacic-Mirski, Thomas Münzel, Andreas Daiber, Moritz Brandt, Tommaso Gori, Hendrik Milting, Stefan Tenzer, Wolfram Ruf, Philip Wenzel
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Research Article Immunology

Targeting myeloid cell coagulation signaling blocks MAP kinase/TGF-β1–driven fibrotic remodeling in ischemic heart failure

Abstract

Despite major advances in acute interventions for myocardial infarction (MI), adverse cardiac remodeling and excess fibrosis after MI causing ischemic heart failure (IHF) remain a leading cause of death worldwide. Here we identify a profibrotic coagulation signaling pathway that can be targeted for improved cardiac function following MI with persistent ischemia. Quantitative phosphoproteomics of cardiac tissue revealed an upregulated mitogen-activated protein kinase (MAPK) pathway in human IHF. Intervention in this pathway with trametinib improves myocardial function and prevents fibrotic remodeling in a murine model of non-reperfused MI. MAPK activation in MI requires myeloid cell signaling of protease-activated receptor 2 linked to the cytoplasmic domain of the coagulation initiator tissue factor (TF). They act upstream of pro-oxidant NOX2 NADPH oxidase, ERK1/2 phosphorylation, and activation of profibrotic TGF-β1. Specific targeting with the TF inhibitor nematode anticoagulant protein c2 (NAPc2) starting 1 day after established experimental MI averts IHF. Increased TF cytoplasmic domain phosphorylation in circulating monocytes from patients with subacute MI identifies a potential thromboinflammatory biomarker reflective of increased risk for IHF and suitable for patient selection to receive targeted TF inhibition therapy.

Authors

Venkata Garlapati, Michael Molitor, Thomas Michna, Gregory S. Harms, Stefanie Finger, Rebecca Jung, Jeremy Lagrange, Panagiotis Efentakis, Johannes Wild, Maike Knorr, Susanne Karbach, Sabine Wild, Ksenija Vujacic-Mirski, Thomas Münzel, Andreas Daiber, Moritz Brandt, Tommaso Gori, Hendrik Milting, Stefan Tenzer, Wolfram Ruf, Philip Wenzel

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Figure 6

Myeloid cell–derived TF-PAR2 complex is required for TGF-β1 activation.

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Myeloid cell–derived TF-PAR2 complex is required for TGF-β1 activation. ...
(A) Confocal microscopy of myocardial cryosections obtained from n = 5 sham-operated and n = 5 LAD-ligated WT (C57BL/6J) mice at day 7. Representative images and quantification of TF+ cells costained for CD45. Unpaired, 2-sided t test. Scale bar: 50 μm. (B–D) TFfl/fl LysMCre and TFfl/fl littermates were subjected to permanent LAD ligation versus sham surgery and investigated after 7 days; n = 5–7 animals per group. (B) Protein expression analysis of p-ERK1/2 (normalized to total ERK1/2) in the infarcted myocardium. Ordinary 1-way ANOVA, Šidák’s multiple-comparison test. (C) Western blot analysis of activated TGF-β1 (normalized to GAPDH) and p-SMAD2 (normalized to total SMAD2) in the infarcted myocardium obtained from TFfl/fl LysMCre and TFfl/fl littermates. Representative blots and quantification of biological replicates. (D) High-frequency ultrasound echocardiography obtained from TFfl/fl LysMCre and TFfl/fl littermates. Ordinary 1-way ANOVA, Šidák’s multiple-comparison test. (E) Sirius red staining and deconvoluted images of fibrotic area on paraffin-embedded heart sections 4 weeks after permanent LAD ligation to induce IHF versus sham surgery. Representative images and quantification of fibrotic areas normalized to surface area. Ordinary 1-way ANOVA, Šidák’s multiple-comparison test; n = 5 animals per group. (F) Kaplan-Meier survival analysis of permanently LAD-ligated TFfl/fl LysMCre and TFfl/fl littermate mice over 4 weeks. Log-rank (Mantel-Cox) test; n = 10–15 animals per group. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001.