Pharmacoidea Ltd., H-6726 Szeged, Hungary.
Boeckeler Instruments, Inc., Tucson, AZ 85714, USA.
Int J Mol Sci. 2022 Jul 9;23(14):7609. doi: 10.3390/ijms23147609.
Despite the growing list of identified SARS-CoV-2 receptors, the human angiotensin-converting enzyme 2 (ACE2) is still viewed as the main cell entry receptor mediating SARS-CoV-2 internalization. It has been reported that wild-type mice, like other rodent species of the Muridae family, cannot be infected with SARS-CoV-2 due to differences in their ACE2 receptors. On the other hand, the consensus heparin-binding motif of SARS-CoV-2's spike protein, PRRAR, enables the attachment to rodent heparan sulfate proteoglycans (HSPGs), including syndecans, a transmembrane HSPG family with a well-established role in clathrin- and caveolin-independent endocytosis. As mammalian syndecans possess a relatively conserved structure, we analyzed the cellular uptake of inactivated SARS-CoV-2 particles in in vitro and in vivo mice models. Cellular studies revealed efficient uptake into murine cell lines with established syndecan-4 expression. After intravenous administration, inactivated SARS-CoV-2 was taken up by several organs in vivo and could also be detected in the brain. Internalized by various tissues, inactivated SARS-CoV-2 raised tissue TNF-α levels, especially in the heart, reflecting the onset of inflammation. Our studies on in vitro and in vivo mice models thus shed light on unknown details of SARS-CoV-2 internalization and help broaden the understanding of the molecular interactions of SARS-CoV-2.
尽管已鉴定出越来越多的 SARS-CoV-2 受体,但人类血管紧张素转换酶 2(ACE2)仍然被视为介导 SARS-CoV-2 内化的主要细胞进入受体。据报道,由于 ACE2 受体的差异,野生型小鼠(像其他鼠科动物)不能感染 SARS-CoV-2。另一方面,SARS-CoV-2 刺突蛋白的肝素结合基序 PRRAR 使病毒能够附着在鼠类肝素硫酸蛋白聚糖(HSPG)上,包括连接蛋白,这是一种在网格蛋白和小窝蛋白非依赖性内吞作用中有明确作用的跨膜 HSPG 家族。由于哺乳动物连接蛋白具有相对保守的结构,我们分析了在体外和体内小鼠模型中失活的 SARS-CoV-2 颗粒的细胞摄取。细胞研究显示,在具有稳定表达的连接蛋白-4 的鼠系细胞中,可有效摄取失活的 SARS-CoV-2。静脉注射后,失活的 SARS-CoV-2 在体内被几个器官摄取,并且在大脑中也可以检测到。失活的 SARS-CoV-2 被各种组织内化,导致组织 TNF-α 水平升高,尤其是在心脏,反映出炎症的发生。因此,我们在体外和体内小鼠模型中的研究揭示了 SARS-CoV-2 内化的未知细节,并有助于加深对 SARS-CoV-2 分子相互作用的理解。