Department of Surgery, Amsterdam University Medical Centers, Amsterdam Cardiovascular Sciences, Location Vrije Universiteit (VU) Medical Center and Academic Medical Centre (AMC), the Netherlands (K.B.R., T.A.R.v.M., N.B., K.K.Y.).
Department of Physiology, Amsterdam University Medical Centers, Amsterdam Cardiovascular Sciences, Location VU Medical Center, the Netherlands (K.B.R., T.A.R.v.M., N.B., J.v.d.V., K.K.Y.).
Arterioscler Thromb Vasc Biol. 2024 Oct;44(10):2226-2243. doi: 10.1161/ATVBAHA.124.321087. Epub 2024 Aug 29.
Abdominal aortic aneurysm (AAA) is characterized by weakening and dilatation of the aortic wall in the abdomen. The aim of this study was to gain insight into cell-specific mechanisms involved in AAA pathophysiology by analyzing the (phospho)proteome of vascular smooth muscle cells derived from patients with AAA compared with those of healthy donors.
A (phospho)proteomics analysis based on tandem mass spectrometry was performed on vascular smooth muscle cells derived from patients with AAA (n=24) and healthy, control individuals (C-SMC, n=8). Following protein identification and quantification using MaxQuant, integrative inferred kinase activity analysis was used to calculate kinase activity scores.
Expression differences between vascular smooth muscle cells derived from patients with AAA and healthy, control individuals were predominantly found in proteins involved in ECM (extracellular matrix) remodeling (THSD4 [thrombospondin type-1 domain-containing protein 4] and ADAMTS1 [A disintegrin and metalloproteinase with thrombospondin motifs 1]), energy metabolism (GYS1 [glycogen synthase 1] and PCK2 [phosphoenolpyruvate carboxykinase 2, mitochondrial]), and contractility (CACNA2D1 [calcium voltage-dependent channel subunit α-2/δ-1] and TPM1 [tropomyosin α-1 chain]). Phosphorylation patterns on proteins related to actin cytoskeleton organization dominated the phosphoproteome of vascular smooth muscle cells derived from patients with AAA . Besides, phosphorylation changes on proteins related to energy metabolism (GYS1), contractility (PARVA [α-parvin], PPP1R12A [protein phosphatase 1 regulatory subunit 12A], and CALD1 [caldesmon 1]), and intracellular communication (GJA1 [gap junction α-1 protein]) were seen. Kinase activity of NUAK1 (NUAK family SNF1-like kinase 1), FYN (tyrosine-protein kinase Fyn), MAPK7 (mitogen-activated protein kinase 7), and STK10 (serine/threonine kinase 10) was different in vascular smooth muscle cells derived from patients with AAA compared with those from healthy, control individuals.
This study revealed changes in expression and phosphorylation levels of proteins involved in various processes responsible for AAA progression and development (eg, energy metabolism, ECM remodeling, actin cytoskeleton organization, contractility, intracellular communication, and cell adhesion). These newly identified proteins, phosphosites, and related kinases provide further insight into the underlying mechanism of vascular smooth muscle cell dysfunction within the aneurysmal wall. Our omics data thereby offer the opportunity to study the relevance, either as drug target or biomarker, of these proteins in AAA development.
腹主动脉瘤（AAA）的特征是腹部主动脉壁的弱化和扩张。本研究的目的是通过分析来自 AAA 患者的血管平滑肌细胞与健康供体的血管平滑肌细胞的（磷酸化）蛋白质组，深入了解 AAA 病理生理学中的细胞特异性机制。
采用基于串联质谱的（磷酸化）蛋白质组学分析方法，对 24 例 AAA 患者来源的血管平滑肌细胞（AAA-SMC）和 8 例健康对照个体来源的血管平滑肌细胞（C-SMC）进行分析。使用 MaxQuant 进行蛋白质鉴定和定量后，采用整合推断的激酶活性分析计算激酶活性评分。
AAA 患者来源的血管平滑肌细胞与健康对照个体来源的血管平滑肌细胞之间的表达差异主要存在于细胞外基质（ECM）重塑（THSD4[血栓素型-1 域包含蛋白 4]和 ADAMTS1[A 型纤溶酶原激活物抑制剂 1 金属蛋白酶 1]）、能量代谢（GYS1[糖原合酶 1]和 PCK2[磷酸烯醇丙酮酸羧激酶 2，线粒体]）和收缩性（CACNA2D1[钙电压依赖性通道亚基α-2/δ-1]和 TPM1[原肌球蛋白α-1 链]）相关蛋白。AAA 患者来源的血管平滑肌细胞的磷酸化蛋白质组主要由与肌动蛋白细胞骨架组织相关的蛋白质的磷酸化模式主导。此外，还观察到与能量代谢（GYS1）、收缩性（PARVA[α-辅肌动蛋白]、PPP1R12A[蛋白磷酸酶 1 调节亚基 12A]和 CALD1[钙调蛋白 1]）和细胞内通讯（GJA1[间隙连接蛋白α-1 蛋白]）相关的蛋白质的磷酸化变化。与健康对照个体来源的血管平滑肌细胞相比，AAA 患者来源的血管平滑肌细胞中 NUAK1（NUAK 家族 SNF1 样激酶 1）、FYN（酪氨酸蛋白激酶 Fyn）、MAPK7（有丝分裂原激活蛋白激酶 7）和 STK10（丝氨酸/苏氨酸激酶 10）的激酶活性存在差异。
本研究揭示了与 AAA 进展和发展相关的各种过程中涉及的蛋白质的表达和磷酸化水平的变化（例如，能量代谢、ECM 重塑、肌动蛋白细胞骨架组织、收缩性、细胞内通讯和细胞黏附）。这些新鉴定的蛋白质、磷酸化位点和相关激酶为血管平滑肌细胞在动脉瘤壁中的功能障碍的潜在机制提供了进一步的认识。我们的组学数据从而提供了研究这些蛋白质在 AAA 发展中的相关性（无论是作为药物靶点还是生物标志物）的机会。
