BACKGROUND:Epsin endocytic adaptor proteins are implicated in the progression of atherosclerosis; however, the underlying molecular mechanisms have not yet been fully defined. In this study, we determined how epsins enhance endothelial-to-mesenchymal transition (EndoMT) in atherosclerosis and assessed the efficacy of a therapeutic peptide in a preclinical model of this disease.METHODS:Using single-cell RNA sequencing combined with molecular, cellular, and biochemical analyses, we investigated the role of epsins in stimulating EndoMT using knockout in Apoe^−/− and lineage tracing/proprotein convertase subtilisin/kexin type 9 serine protease mutant viral-induced atherosclerotic mouse models. The therapeutic efficacy of a synthetic peptide targeting atherosclerotic plaques was then assessed in Apoe^−/− mice.RESULTS:Single-cell RNA sequencing and lineage tracing revealed that epsins 1 and 2 promote EndoMT and that the loss of endothelial epsins inhibits EndoMT marker expression and transforming growth factor-β signaling in vitro and in atherosclerotic mice, which is associated with smaller lesions in the Apoe^−/− mouse model. Mechanistically, the loss of endothelial cell epsins results in increased fibroblast growth factor receptor-1 expression, which inhibits transforming growth factor-β signaling and EndoMT. Epsins directly bind ubiquitinated fibroblast growth factor receptor-1 through their ubiquitin-interacting motif, which results in endocytosis and degradation of this receptor complex. Consequently, administration of a synthetic ubiquitin-interacting motif–containing peptide atheroma ubiquitin-interacting motif peptide inhibitor significantly attenuates EndoMT and progression of atherosclerosis.CONCLUSIONS:We conclude that epsins potentiate EndoMT during atherogenesis by increasing transforming growth factor-β signaling through fibroblast growth factor receptor-1 internalization and degradation. Inhibition of EndoMT by reducing epsin–fibroblast growth factor receptor-1 interaction with a therapeutic peptide may represent a novel treatment strategy for atherosclerosis.

中文翻译：

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靶向 Epsins 抑制成纤维细胞生长因子信号传导，同时增强转化生长因子-β 信号传导限制动脉粥样硬化中的内皮细胞向间质转化

背景：Epsin 内吞衔接蛋白与动脉粥样硬化的进展有关。然而，潜在的分子机制尚未完全明确。在这项研究中，我们确定了epsins如何增强动脉粥样硬化中的内皮间质转化（EndoMT），并评估了治疗肽在该疾病的临床前模型中的功效。方法：使用单细胞RNA测序结合分子、细胞、和生化分析，我们使用Apoe ^−/−敲除和谱系追踪/前蛋白转化酶枯草杆菌蛋白酶/kexin 9 型丝氨酸蛋白酶突变体病毒诱导的动脉粥样硬化小鼠模型研究了 epsins 在刺激 EndoMT 中的作用。然后在Apoe ^−/−小鼠中评估了针对动脉粥样硬化斑块的合成肽的治疗效果。 结果：单细胞 RNA 测序和谱系追踪显示，epsins 1 和 2 促进 EndoMT，内皮 epsins 的缺失会抑制 EndoMT 标记物的表达，并且在体外和动脉粥样硬化小鼠中转化生长因子-β 信号，这与Apoe ^−/−小鼠模型中较小的病变有关。从机制上讲，内皮细胞 Epsins 的缺失会导致成纤维细胞生长因子受体 1 表达增加，从而抑制转化生长因子 β 信号传导和 EndoMT。Epsins 通过其泛素相互作用基序直接结合泛素化成纤维细胞生长因子受体 1，从而导致该受体复合物的内吞作用和降解。因此，使用合成的含有泛素相互作用基序的肽粥样斑块泛素相互作用基序肽抑制剂可显着减弱 EndoMT 和动脉粥样硬化的进展。 结论：我们得出结论，epsins 通过通过成纤维细胞生长因子增加转化生长因子-β 信号传导，在动脉粥样硬化形成过程中增强 EndoMT受体1内化和降解。通过减少epsin-成纤维细胞生长因子受体1与治疗肽的相互作用来抑制EndoMT可能代表了动脉粥样硬化的一种新的治疗策略。