Institute for Cardiogenetics, University of Luebeck, Luebeck, Germany.
DZHK (German Research Centre for Cardiovascular Research) Partner Site Hamburg/Luebeck/Kiel, Luebeck, Germany.
Sci Rep. 2024 May 28;14(1):12212. doi: 10.1038/s41598-024-62878-3.
A positive family history is a major independent risk factor for atherosclerosis, and genetic variation is an important aspect of cardiovascular disease research. We identified a heterozygous missense variant p.L245P in the MMP10 gene in two families with premature myocardial infarction using whole-exome sequencing. The aim of this study was to investigate the consequences of this variant using in-silico and functional in-vitro assays. Molecular dynamics simulations were used to analyze protein interactions, calculate free binding energy, and measure the volume of the substrate-binding cleft of MMP10-TIMP1 models. The p.L245P variant showed an altered protein surface, different intra- and intermolecular interactions of MMP10-TIMP1, a lower total free binding energy between MMP10-TIMP1, and a volume-minimized substrate-binding cleft of MMP10 compared to the wild-type. For the functional assays, human THP-1 cells were transfected with plasmids containing MMP10 cDNA carrying the p.L245P and wild-type variant and differentiated into macrophages. Macrophage adhesion and migration assays were then conducted, and pro-inflammatory chemokine levels were evaluated. The p.L245P variant led to macrophages that were more adherent, less migratory, and secreted higher levels of the pro-inflammatory chemokines CXCL1 and CXCL8 than wild-type macrophages. Thus, the p.L245P variant in MMP10 may influence the pathogenesis of atherosclerosis in families with premature myocardial infarction by altering protein - protein interactions, macrophage adhesion and migration, and expression of pro-inflammatory chemokines, which may increase plaque rupture. These results could contribute to the development of selective MMP10 inhibitors and reduce the risk of atherosclerosis in families with a history of premature myocardial infarction.
阳性家族史是动脉粥样硬化的一个主要独立危险因素,遗传变异是心血管疾病研究的一个重要方面。我们通过全外显子组测序在两个早发心肌梗死的家族中发现了 MMP10 基因中的杂合错义变体 p.L245P。本研究旨在通过计算机模拟和体外功能实验来研究该变体的后果。使用分子动力学模拟分析蛋白相互作用,计算自由结合能,并测量 MMP10-TIMP1 模型的底物结合裂隙体积。与野生型相比,p.L245P 变体显示出改变的蛋白表面、MMP10-TIMP1 不同的内部分子和分子间相互作用、MMP10-TIMP1 之间的总自由结合能降低以及 MMP10 的底物结合裂隙体积最小化。对于功能实验,将含有携带 p.L245P 和野生型变体的 MMP10 cDNA 的质粒转染人 THP-1 细胞并分化为巨噬细胞。然后进行巨噬细胞黏附和迁移实验,并评估促炎趋化因子水平。与野生型巨噬细胞相比,p.L245P 变体导致巨噬细胞黏附性增加、迁移性降低且分泌更高水平的促炎趋化因子 CXCL1 和 CXCL8。因此,MMP10 中的 p.L245P 变体可能通过改变蛋白-蛋白相互作用、巨噬细胞黏附和迁移以及促炎趋化因子的表达来影响早发心肌梗死家族中动脉粥样硬化的发病机制,从而增加斑块破裂的风险。这些结果可能有助于开发选择性 MMP10 抑制剂,并降低有早发心肌梗死家族史的个体发生动脉粥样硬化的风险。