Table1_SARS-CoV-2 Employ BSG/CD147 and ACE2 Receptors to Directly Infect Human Induced Pluripotent Stem Cell-Derived Kidney Podocytes.XLSX

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes the Coronavirus disease 2019 (COVID-19), which has resulted in over 5.9 million deaths worldwide. While cells in the respiratory system are the initial target of SARS-CoV-2, there is mounting evidence that COVID-19 is a multi-organ disease. Still, the direct affinity of SARS-CoV-2 for cells in other organs such as the kidneys, which are often targeted in severe COVID-19, remains poorly understood. We employed a human induced pluripotent stem (iPS) cell-derived model to investigate the affinity of SARS-CoV-2 for kidney glomerular podocytes, and examined the expression of host factors for binding and processing of the virus. We studied cellular uptake of the live SARS-CoV-2 virus as well as a pseudotyped virus. Infection of podocytes with live SARS-CoV-2 or spike-pseudotyped lentiviral particles revealed cellular uptake even at low multiplicity of infection (MOI) of 0.01. We found that direct infection of human iPS cell-derived podocytes by SARS-CoV-2 virus can cause cell death and podocyte foot process retraction, a hallmark of podocytopathies and progressive glomerular diseases including collapsing glomerulopathy observed in patients with severe COVID-19 disease. We identified BSG/CD147 and ACE2 receptors as key mediators of spike binding activity in human iPS cell-derived podocytes. These results show that SARS-CoV-2 can infect kidney glomerular podocytes in vitro via multiple binding interactions and partners, which may underlie the high affinity of SARS-CoV-2 for kidney tissues. This stem cell-derived model is potentially useful for kidney-specific antiviral drug screening and mechanistic studies of COVID-19 organotropism.

严重急性呼吸综合征冠状病毒2（SARS-CoV-2）可引发新型冠状病毒肺炎（COVID-19），该疾病已在全球造成超过590万例死亡。尽管呼吸系统细胞是SARS-CoV-2的初始感染靶点，但越来越多的证据表明，新型冠状病毒肺炎是一种多器官疾病。然而，SARS-CoV-2对重症COVID-19患者常累及的肾脏等其他器官细胞的直接亲和力，目前仍不甚明确。本研究采用人类诱导多能干细胞（iPS）衍生模型，探究SARS-CoV-2对肾脏肾小球足细胞的亲和力，并检测了病毒结合与加工过程所需的宿主因子表达水平。我们分别研究了活SARS-CoV-2与假型病毒的细胞摄取情况。研究发现，即使用感染复数（MOI）低至0.01的活SARS-CoV-2或刺突蛋白假型慢病毒颗粒感染足细胞，仍可实现细胞摄取。我们发现，SARS-CoV-2直接感染人类iPS细胞衍生的足细胞可引发细胞死亡及足细胞足突回缩——这正是重症COVID-19患者中观察到的足细胞病、塌陷性肾小球病等进行性肾小球疾病的标志性特征。我们确定，BSG/CD147与血管紧张素转换酶2（ACE2）受体是人类iPS细胞衍生足细胞中刺突蛋白结合活性的关键介导因子。上述结果表明，SARS-CoV-2可通过多种结合相互作用及结合伴侣在体外感染肾脏肾小球足细胞，这或许是SARS-CoV-2对肾脏组织具有高亲和力的分子基础。该干细胞衍生模型有望用于肾脏特异性抗病毒药物筛选以及COVID-19器官嗜性的机制研究。