Abstract：
UNASSIGNED: Cell phenotype switching is increasingly being recognized in atherosclerosis. However, our understanding of the exact stimuli for such cellular transformations and their significance for human atherosclerosis is still evolving. Intraplaque hemorrhage is thought to be a major contributor to plaque progression in part by stimulating the influx of CD163+ macrophages. Here, we explored the hypothesis that CD163+ macrophages cause plaque progression through the induction of proapoptotic endothelial-to-mesenchymal transition (EndMT) within the fibrous cap.
UNASSIGNED: Human coronary artery sections from CVPath\'s autopsy registry were selected for pathological analysis. Athero-prone ApoE-/- and ApoE-/-/CD163-/- mice were used for in vivo studies. Human peripheral blood mononuclear cell-induced macrophages and human aortic endothelial cells were used for in vitro experiments.
UNASSIGNED: In 107 lesions with acute coronary plaque rupture, 55% had pathological evidence of intraplaque hemorrhage in nonculprit vessels/lesions. Thinner fibrous cap, greater CD163+ macrophage accumulation, and a larger number of CD31/FSP-1 (fibroblast specific protein-1) double-positive cells and TUNEL (terminal deoxynucleotidyl transferase-dUTP nick end labeling) positive cells in the fibrous cap were observed in nonculprit intraplaque hemorrhage lesions, as well as in culprit rupture sections versus nonculprit fibroatheroma sections. Human aortic endothelial cells cultured with supernatants from hemoglobin/haptoglobin-exposed macrophages showed that increased mesenchymal marker proteins (transgelin and FSP-1) while endothelial markers (VE-cadherin and CD31) were reduced, suggesting EndMT induction. Activation of NF-κB (nuclear factor kappa β) signaling by proinflammatory cytokines released from CD163+ macrophages directly regulated the expression of Snail, a critical transcription factor during EndMT induction. Western blot analysis for cleaved caspase-3 and microarray analysis of human aortic endothelial cells indicated that apoptosis was stimulated during CD163+ macrophage-induced EndMT. Additionally, CD163 deletion in athero-prone mice suggested that CD163 is required for EndMT and plaque progression. Using single-cell RNA sequencing from human carotid endarterectomy lesions, a population of EndMT was detected, which demonstrated significant upregulation of apoptosis-related genes.
UNASSIGNED: CD163+ macrophages provoke EndMT, which may promote plaque progression through fibrous cap thinning.
UNASSIGNED: Human coronary artery sections from CVPath\'s autopsy registry were selected for pathological analysis. Athero-prone ApoE-/- and ApoE-/-/CD163-/- mice were used for in vivo studies. Human peripheral blood mononuclear cell-induced macrophages and human aortic endothelial cells were used for in vitro experiments.
UNASSIGNED: In 107 lesions with acute coronary plaque rupture, 55% had pathological evidence of intraplaque hemorrhage in nonculprit vessels/lesions. Thinner fibrous cap, greater CD163+ macrophage accumulation, and a larger number of CD31/FSP-1 (fibroblast specific protein-1) double-positive cells and TUNEL (terminal deoxynucleotidyl transferase-dUTP nick end labeling) positive cells in the fibrous cap were observed in nonculprit intraplaque hemorrhage lesions, as well as in culprit rupture sections versus nonculprit fibroatheroma sections. Human aortic endothelial cells cultured with supernatants from hemoglobin/haptoglobin-exposed macrophages showed that increased mesenchymal marker proteins (transgelin and FSP-1) while endothelial markers (VE-cadherin and CD31) were reduced, suggesting EndMT induction. Activation of NF-κB (nuclear factor kappa β) signaling by proinflammatory cytokines released from CD163+ macrophages directly regulated the expression of Snail, a critical transcription factor during EndMT induction. Western blot analysis for cleaved caspase-3 and microarray analysis of human aortic endothelial cells indicated that apoptosis was stimulated during CD163+ macrophage-induced EndMT. Additionally, CD163 deletion in athero-prone mice suggested that CD163 is required for EndMT and plaque progression. Using single-cell RNA sequencing from human carotid endarterectomy lesions, a population of EndMT was detected, which demonstrated significant upregulation of apoptosis-related genes.
UNASSIGNED: CD163+ macrophages provoke EndMT, which may promote plaque progression through fibrous cap thinning.
摘要：
■在动脉粥样硬化中细胞表型转换越来越被认识到。然而,我们对这种细胞转化的确切刺激及其对人类动脉粥样硬化的意义的理解仍在不断发展。斑块内出血被认为是部分通过刺激CD163+巨噬细胞的流入而导致斑块进展的主要因素。这里,我们探索了以下假设:CD163巨噬细胞通过诱导纤维帽内的促凋亡内皮-间质转化(EndMT)引起斑块进展.
■选择CVPath尸检注册表中的人冠状动脉切片进行病理分析。将有动脉粥样硬化倾向的ApoE-/-和ApoE-/-/CD163-/-小鼠用于体内研究。人外周血单核细胞诱导的巨噬细胞和人主动脉内皮细胞用于体外实验。
■在107个急性冠状动脉斑块破裂的病变中，55％的非罪犯血管/病变中有斑块内出血的病理证据。更薄的纤维帽，更大的CD163+巨噬细胞积累，在非罪犯斑块内出血病变中观察到纤维帽中大量的CD31/FSP-1（成纤维细胞特异性蛋白-1）双阳性细胞和TUNEL阳性细胞，以及罪犯破裂切片与非罪犯纤维粥样硬化切片。用血红蛋白/触珠蛋白暴露的巨噬细胞的上清液培养的人主动脉内皮细胞显示，间充质标记蛋白（transgelin和FSP-1）增加，而内皮标记（VE-cadherin和CD31）减少，暗示EndMT诱导。CD163+巨噬细胞释放的促炎细胞因子激活NF-κB（核因子κβ）信号直接调节Snail的表达，EndMT诱导过程中的关键转录因子。对裂解的半胱天冬酶3的Western印迹分析和人主动脉内皮细胞的微阵列分析指示在CD163+巨噬细胞诱导的EndMT期间刺激细胞凋亡。此外,动脉粥样硬化倾向小鼠中的CD163缺失表明CD163是EndMT和斑块进展所必需的。使用人颈动脉内膜切除术病变的单细胞RNA测序，检测到一个EndMT群体，这表明凋亡相关基因显著上调。
■CD163+巨噬细胞引起EndMT，这可能通过纤维帽变薄促进斑块进展。
■选择CVPath尸检注册表中的人冠状动脉切片进行病理分析。将有动脉粥样硬化倾向的ApoE-/-和ApoE-/-/CD163-/-小鼠用于体内研究。人外周血单核细胞诱导的巨噬细胞和人主动脉内皮细胞用于体外实验。
■在107个急性冠状动脉斑块破裂的病变中，55％的非罪犯血管/病变中有斑块内出血的病理证据。更薄的纤维帽，更大的CD163+巨噬细胞积累，在非罪犯斑块内出血病变中观察到纤维帽中大量的CD31/FSP-1（成纤维细胞特异性蛋白-1）双阳性细胞和TUNEL阳性细胞，以及罪犯破裂切片与非罪犯纤维粥样硬化切片。用血红蛋白/触珠蛋白暴露的巨噬细胞的上清液培养的人主动脉内皮细胞显示，间充质标记蛋白（transgelin和FSP-1）增加，而内皮标记（VE-cadherin和CD31）减少，暗示EndMT诱导。CD163+巨噬细胞释放的促炎细胞因子激活NF-κB（核因子κβ）信号直接调节Snail的表达，EndMT诱导过程中的关键转录因子。对裂解的半胱天冬酶3的Western印迹分析和人主动脉内皮细胞的微阵列分析指示在CD163+巨噬细胞诱导的EndMT期间刺激细胞凋亡。此外,动脉粥样硬化倾向小鼠中的CD163缺失表明CD163是EndMT和斑块进展所必需的。使用人颈动脉内膜切除术病变的单细胞RNA测序，检测到一个EndMT群体，这表明凋亡相关基因显著上调。
■CD163+巨噬细胞引起EndMT，这可能通过纤维帽变薄促进斑块进展。
