组织蛋白酶 S 是整合素 αvβ3 触发血管退行性变、夹层和破裂的内源性调节剂。
Legumain Is an Endogenous Modulator of Integrin αvβ3 Triggering Vascular Degeneration, Dissection, and Rupture.
机构信息
Institutes of Biomedical Sciences, Fudan University, Shanghai, China (L.P., S.C., A.S., J.G.).
Key Laboratory of Viral Heart Diseases, National Health Commission, Shanghai, China (L.P., P.B., X.W., J.L., S.C., X.M., A.S., J.G.).
出版信息
Circulation. 2022 Mar;145(9):659-674. doi: 10.1161/CIRCULATIONAHA.121.056640. Epub 2022 Jan 31.
BACKGROUND
The development of thoracic aortic dissection (TAD) is closely related to extracellular matrix degradation and vascular smooth muscle cell (VSMC) transformation from contractile to synthetic type. LGMN (legumain) degrades extracellular matrix components directly or by activating downstream signals. The role of LGMN in VSMC differentiation and the occurrence of TAD remains elusive.
METHODS
Microarray datasets concerning vascular dissection or aneurysm were downloaded from the Gene Expression Omnibus database to screen differentially expressed genes. Four-week-old male knockout mice (Lgmn), macrophage-specific knockout mice (Lgmn;LysM), and RR-11a-treated C57BL/6 mice were given BAPN (β-aminopropionitrile monofumarate; 1 g/kg/d) in drinking water for 4 weeks for TAD modeling. RNA sequencing analysis was performed to recapitulate transcriptome profile changes. Cell interaction was examined in macrophage and VSMC coculture system. The reciprocity of macrophage-derived LGMN with integrin αvβ3 in VSMCs was tested by coimmunoprecipitation assay and colocalization analyses.
RESULTS
Microarray datasets from the Gene Expression Omnibus database indicated upregulated LGMN in aorta from patients with TAD and mice with angiotensin II-induced AAA. Elevated LGMN was evidenced in aorta and sera from patients with TAD and mice with BAPN-induced TAD. BAPN-induced TAD progression was significantly ameliorated in Lgmn-deficient or inhibited mice. Macrophage-specific deletion of alleviated BAPN-induced extracellular matrix degradation. Unbiased profiler polymerase chain reaction array and Gene Ontology analysis displayed that regulated VSMC phenotype transformation. Macrophage-specific deletion of ameliorated VSMC phenotypic switch in BAPN-treated mice. Macrophage-derived LGMN inhibited VSMC differentiation in vitro as assessed by macrophages and the VSMC coculture system. Macrophage-derived LGMN bound to integrin αvβ3 in VSMCs and blocked integrin αvβ3, thereby attenuating Rho GTPase activation, downregulating VSMC differentiation markers and eventually exacerbating TAD development. ROCK (Rho kinase) inhibitor Y-27632 reversed the protective role of LGMN depletion in vascular dissection.
CONCLUSIONS
LGMN signaling may be a novel target for the prevention and treatment of TAD.
背景
胸主动脉夹层（TAD）的发生发展与细胞外基质降解和血管平滑肌细胞（VSMC）从收缩型向合成型转化密切相关。LGMN（溶酶体半胱氨酸蛋白酶）通过直接降解细胞外基质成分或激活下游信号来发挥作用。LGMN 在 VSMC 分化和 TAD 发生中的作用仍不清楚。
方法
从基因表达综合数据库（GEO）中下载与血管夹层或动脉瘤相关的微阵列数据集，以筛选差异表达基因。4 周龄雄性 基因敲除（Lgmn）小鼠、巨噬细胞特异性 基因敲除（Lgmn；LysM）小鼠和 RR-11a 处理的 C57BL/6 小鼠给予 BAPN（β-氨基丙腈单富马酸盐；1 g/kg/d）饮用水 4 周，建立 TAD 模型。通过 RNA 测序分析重现转录组谱变化。在巨噬细胞和 VSMC 共培养系统中检测细胞相互作用。通过免疫共沉淀和共定位分析检测巨噬细胞来源的 LGMN 与 VSMC 整合素 αvβ3 的相互作用。
结果
GEO 数据库中的微阵列数据集表明，TAD 患者和血管紧张素 II 诱导的 AAA 小鼠的主动脉中 LGMN 上调。TAD 患者的主动脉和血清中也检测到 LGMN 升高，BAPN 诱导的 TAD 小鼠也检测到 LGMN 升高。Lgmn 缺陷或抑制小鼠的 BAPN 诱导的 TAD 进展明显改善。巨噬细胞特异性 缺失减轻了 BAPN 诱导的细胞外基质降解。无偏置分析聚合酶链反应（PCR）阵列和基因本体论分析显示， 调节 VSMC 表型转化。巨噬细胞特异性 缺失改善了 BAPN 处理小鼠的 VSMC 表型转换。通过巨噬细胞和 VSMC 共培养系统评估，巨噬细胞衍生的 LGMN 抑制了体外 VSMC 分化。巨噬细胞衍生的 LGMN 与 VSMCs 中的整合素 αvβ3 结合，阻断整合素 αvβ3，从而抑制 Rho GTPase 激活，下调 VSMC 分化标志物，最终加重 TAD 发展。Rho 激酶（ROCK）抑制剂 Y-27632 逆转了 LGMN 耗竭在血管夹层中的保护作用。
结论
LGMN 信号可能是预防和治疗 TAD 的一个新靶点。
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