Center for Thrombosis and Hemostasis (V.G., Q.L., J.P., M.A., T.S.N., K.G., M.M., S.F., W.R., P.W.), University Medical Center Mainz, Germany.
Department of Cardiology (V.G., Q.L., M.A., M.M., S.F., P.W.), University Medical Center Mainz, Germany.
Circ Res. 2024 Sep 27;135(8):841-855. doi: 10.1161/CIRCRESAHA.123.324114. Epub 2024 Sep 5.
Excess fibrotic remodeling causes cardiac dysfunction in ischemic heart disease, driven by MAP (mitogen-activated protein) kinase-dependent TGF-ß1 (transforming growth factor-ß1) activation by coagulation signaling of myeloid cells. How coagulation-inflammatory circuits can be specifically targeted to achieve beneficial macrophage reprogramming after myocardial infarction (MI) is not completely understood.
Mice with permanent ligation of the left anterior descending artery were used to model nonreperfused MI and analyzed by single-cell RNA sequencing, protein expression changes, confocal microscopy, and longitudinal monitoring of recovery. We probed the role of the tissue factor (TF)-FVIIa (activated factor VII)-integrin ß1-PAR2 (protease-activated receptor 2) signaling complex by utilizing genetic mouse models and pharmacological intervention.
Cleavage-insensitive PAR2 and myeloid cell integrin ß1-deficient mice had improved cardiac function after MI compared with controls. Proximity ligation assays of monocytic cells demonstrated that colocalization of FVIIa with integrin ß1 was diminished in monocyte/macrophage FVII-deficient mice after MI. Compared with controls, F7 CX3CR1 (CX3C motif chemokine receptor 1) mice showed reduced TGF-ß1 and MAP kinase activation, as well as cardiac dysfunction after MI, despite unaltered overall recruitment of myeloid cells. Single-cell mRNA sequencing of CD45 (cluster of differentiation 45) cells 3 and 7 days after MI uncovered a trajectory from recruited monocytes to inflammatory TF/TREM (triggered receptor expressed on myeloid cells) 1 macrophages requiring F7. As early as 7 days after MI, macrophage F7 deletion led to an expansion of reparative Olfml 3 (olfactomedin-like protein 3) macrophages and, conversely, to a reduction of TF/TREM1 macrophages, which were also reduced in PAR2 mice. Short-term treatment from days 1 to 5 after nonreperfused MI with a monoclonal antibody inhibiting the macrophage TF-FVIIa-PAR2 signaling complex without anticoagulant activity improved cardiac dysfunction, decreased excess fibrosis, attenuated vascular endothelial dysfunction, and increased survival 28 days after MI.
Extravascular TF-FVIIa-PAR2 complex signaling drives inflammatory macrophage polarization in ischemic heart disease. Targeting this signaling complex for specific therapeutic macrophage reprogramming following MI attenuates cardiac fibrosis and improves cardiovascular function.
在缺血性心脏病中，过多的纤维性重塑导致心脏功能障碍，这是由髓样细胞的凝血信号驱动的 MAP（有丝分裂原激活蛋白）激酶依赖性 TGF-β1（转化生长因子-β1）激活引起的。不完全清楚如何特异性地靶向凝血-炎症回路，以实现心肌梗死后（MI）有益的巨噬细胞重编程。
利用左前降支永久性结扎的小鼠模型进行非再灌注 MI，并通过单细胞 RNA 测序、蛋白表达变化、共聚焦显微镜和恢复的纵向监测进行分析。我们利用基因敲除小鼠模型和药理学干预来研究组织因子（TF）-FVIIa（激活的因子 VII）-整合素β1-PAR2（蛋白酶激活受体 2）信号复合物的作用。
与对照组相比，PAR2 切割不敏感和髓样细胞整合素β1 缺陷小鼠在 MI 后心功能得到改善。单核细胞的接近连接测定显示，MI 后单核细胞/巨噬细胞 FVII 缺陷小鼠中 FVIIa 与整合素β1 的共定位减少。与对照组相比，F7 CX3CR1（CX3C 基序趋化因子受体 1）小鼠在 MI 后显示 TGF-β1 和 MAP 激酶激活减少以及心功能障碍，尽管整体募集的髓样细胞没有改变。MI 后 3 天和 7 天 CD45（分化簇 45）细胞的单细胞 mRNA 测序揭示了一条从募集的单核细胞到炎症 TF/TREM（触发受体表达在髓样细胞上）1 巨噬细胞的轨迹，该轨迹需要 F7。早在 MI 后 7 天，巨噬细胞 F7 缺失导致修复性 Olfml3（olfactomedin-like protein 3）巨噬细胞的扩张，相反，TF/TREM1 巨噬细胞减少，PAR2 小鼠中也减少了 TF/TREM1 巨噬细胞。在非再灌注 MI 后 1 至 5 天用单克隆抗体抑制无抗凝活性的巨噬细胞 TF-FVIIa-PAR2 信号复合物进行短期治疗可改善心功能障碍、减少过度纤维化、减轻血管内皮功能障碍，并增加 MI 后 28 天的存活率。
细胞外 TF-FVIIa-PAR2 复合物信号驱动缺血性心脏病中的炎症性巨噬细胞极化。针对 MI 后这种信号复合物进行特定的治疗性巨噬细胞重编程可减轻心肌纤维化并改善心血管功能。
