Bach Marie Lykke, Laftih Sara, Andresen Jesper K, Pedersen Rune M, Andersen Thomas Emil, Madsen Lone W, Madsen Kirsten, Hinrichs Gitte R, Zachar Rikke, Svenningsen Per, Lund Lars, Johansen Isik S, Hansen Lennart Friis, Palarasah Yaseelan, Jensen Boye L
Unit of Cardiovascular and Renal Research, Department of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
Department of Clinical Microbiology, Odense University Hospital, and Research Unit for Clinical Microbiology, University of Southern Denmark, Odense, Denmark.
Pflugers Arch. 2025 Jan;477(1):83-98. doi: 10.1007/s00424-024-03022-y. Epub 2024 Oct 9.
SARS-CoV-2 virus infects cells by engaging with ACE2 requiring protease TMPRSS2. ACE2 is highly expressed in kidneys. Predictors for severe disease are high age and male sex. We hypothesized that ACE2 and TMPRSS2 proteins are more abundant (1) in males and with increasing age in kidney and (2) in urine and extracellular vesicles (EVs) from male patients with COVID-19 and (3) SARS-CoV-2 is present in urine and EVs during infection. Kidney cortex samples from patients subjected to cancer nephrectomy (male/female; < 50 years/˃75 years, n = 24; ˃80 years, n = 15) were analyzed for ACE2 and TMPRSS2 protein levels. Urine from patients hospitalized with SARS-CoV-2 infection was analyzed for ACE2 and TMPRSS2. uEVs were used for immunoblotting and SARS-CoV-2 mRNA and antigen detection. Tissue ACE2 and TMPRSS2 protein levels did not change with age. ACE2 was not more abundant in male kidneys in any age group. ACE2 protein was associated with proximal tubule apical membranes in cortex. TMPRSS2 was observed predominantly in the medulla. ACE2 was elevated significantly in uEVs and urine from patients with COVID-19 with no sex difference compared with urine from controls w/wo albuminuria. TMPRSS2 was elevated in uEVs from males compared to female. ACE2 and TMPRSS2 did not co-localize in uEVs/apical membranes. SARS-CoV-2 nucleoprotein and mRNA were not detected in urine. Higher kidney ACE2 protein abundance is unlikely to explain higher susceptibility to SARS-CoV-2 infection in males. Kidney tubular cells appear not highly susceptible to SARS-CoV-2 infection. Loss of ACE2 into urine in COVID could impact susceptibility and angiotensin metabolism.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）通过与需要蛋白酶TMPRSS2的血管紧张素转换酶2（ACE2）结合来感染细胞。ACE2在肾脏中高度表达。严重疾病的预测因素是高龄和男性。我们假设ACE2和TMPRSS2蛋白（1）在男性中以及在肾脏中随年龄增长而更为丰富，（2）在患有新型冠状病毒肺炎（COVID-19）的男性患者的尿液和细胞外囊泡（EVs）中更为丰富，以及（3）在感染期间SARS-CoV-2存在于尿液和EVs中。对接受癌症肾切除术的患者（男性/女性；<50岁/>75岁，n = 24；>80岁，n = 15）的肾皮质样本进行ACE2和TMPRSS2蛋白水平分析。对因SARS-CoV-2感染住院的患者的尿液进行ACE2和TMPRSS2分析。使用尿细胞外囊泡进行免疫印迹以及SARS-CoV-2 mRNA和抗原检测。组织ACE2和TMPRSS2蛋白水平不随年龄变化。在任何年龄组中，ACE2在男性肾脏中并不更为丰富。ACE2蛋白与皮质中的近端小管顶端膜相关。TMPRSS2主要在髓质中观察到。与有或无蛋白尿的对照患者的尿液相比，COVID-19患者的尿细胞外囊泡和尿液中的ACE2显著升高，且无性别差异。与女性相比，男性尿细胞外囊泡中的TMPRSS2升高。ACE2和TMPRSS2在尿细胞外囊泡/顶端膜中不共定位。尿液中未检测到SARS-CoV-2核蛋白和mRNA。肾脏中较高的ACE2蛋白丰度不太可能解释男性对SARS-CoV-2感染较高的易感性。肾小管细胞似乎对SARS-CoV-2感染不太敏感。COVID患者尿液中ACE2的丢失可能会影响易感性和血管紧张素代谢。
