Soluble ACE2-mediated cell entry of SARS-CoV-2 via interaction with proteins related to the renin-angiotensin system
Authors: Tak Mao Chan
Jasper Fuk-Woo Chan
Sin-Yee Fung
Susanna Kar Pui Lau
Susan Yung
Zongwei Cai
Kwok-Hung Chan
Kelvin Kai-Wang To
Jian-Piao Cai
Runhong Zhou
Patrick Chiu Yat Woo
Kwok-Yung Yuen
Jade Lee Lee Teng
Lin Zhu
Zhiwei Chen
Hanjun Zhao
Chaoyu Zhang
Kam-Leung Siu
Dong-Yan Jin
Lilong Jia
Chengxi Huang
Man Lung Yeung
Journal: Cell (2021)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: ACE2; COVID-19; RNAi; SARS-CoV-2; sACE2
Affiliations: State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Department of Clinical Microbiology and Infection Control, The University of Hong Kong-Shenzhen Hospital, Shenzhen, Guangdong Province, China; Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China. Electronic address: ., State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China., Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China., Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; AIDS Institute, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China., State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Department of Clinical Microbiology and Infection Control, The University of Hong Kong-Shenzhen Hospital, Shenzhen, Guangdong Province, China; Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China., State Key Laboratory of Environmental and Biological Analysis, Department of Chemistry, Hong Kong Baptist University, Hong Kong Special Administrative Region, China., Department of Biochemistry, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China., Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China., State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Department of Clinical Microbiology and Infection Control, The University of Hong Kong-Shenzhen Hospital, Shenzhen, Guangdong Province, China; Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China., State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Department of Clinical Microbiology and Infection Control, The University of Hong Kong-Shenzhen Hospital, Shenzhen, Guangdong Province, China; Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; AIDS Institute, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China., State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China. Electronic address: ., Department of Clinical Microbiology and Infection Control, The University of Hong Kong-Shenzhen Hospital, Shenzhen, Guangdong Province, China; Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China. Electronic address: .

Abstract:  Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can cause acute respiratory disease and multiorgan failure. Finding human host factors that are essential for SARS-CoV-2 infection could facilitate the formulation of treatment strategies. Using a human kidney cell line-HK-2-that is highly susceptible to SARS-CoV-2, we performed a genome-wide RNAi screen and identified virus dependency factors (VDFs), which play regulatory roles in biological pathways linked to clinical manifestations of SARS-CoV-2 infection. We found a role for a secretory form of SARS-CoV-2 receptor, soluble angiotensin converting enzyme 2 (sACE2), in SARS-CoV-2 infection. Further investigation revealed that SARS-CoV-2 exploits receptor-mediated endocytosis through interaction between its spike with sACE2 or sACE2-vasopressin via AT1 or AVPR1B, respectively. Our identification of VDFs and the regulatory effect of sACE2 on SARS-CoV-2 infection shed insight into pathogenesis and cell entry mechanisms of SARS-CoV-2 as well as potential treatment strategies for COVID-19. 

Retracted: No

PubPeer comments:
  Figure 4B from this paper. 1) There are many separate gel pieces including single lane slots pasted together. 2) There are no MW markers to tell the reader the protein sizes. This is problematic because of point 3. 3) The authors report they are using Flag-vasopressin. Vasopressin (AVP) is a small peptide of 1084 MW. The Flag tag, is in a single copy would add 1 Kd. From the STAR Methods:  Recombinant SARS-CoV-2 S proteins (Sino Biological) were incubated with protein lysates expressing V5-tagged or untagged ACE2 and FLAG-tagged vasopressin under previously described conditions... The STAR Methods (Table of Reagents- Recombinant DNA does not list this reagent). What was actually transfected? The coding region of the AVP peptide? The whole precursor protein including vasopressin, neurophysin II, and copeptin? If its the whole precursor what actually binds to ACE2 and how can it bind to the AVPR1? , 
 #1 Thanks for bringing this up, chlamydial parasite of free-living amoebae. I see that the supplementary figures at Mendeley Data are just more of the normal processed figures, with no original data for the Blots or, for that matter, any of the other supplied imagery. Even though: **Mendeley Data Instructions JF - Home: Cell Press:** *u201cIf your paper is accepted, we encourage you to publish the original, unprocessed data alongside your Cell Press paper through Mendeley Data.u201d* And - **Why you should deposit your raw data - Cell:** *u201cThe most important reason to deposit your raw data is to facilitate image screening once your paper is accepted. All accepted manuscripts at Cell Press undergo an image screening process. In this process, technicians (and not editors) examine all images in an accepted paper, looking for evidence of manipulation and for problems with image quality.u201d* Apparently the overworked grunts didnu2019t notice anything untoward but I guess it is more important to rapidly publish COVID-19 papers in Cell than to provide common or garden scientific readers with the underlying data. , 
 Discussed at RetractionWatch: https: retractionwatch.com 2022 10 04 when-failure-to-correct-a-flawed-paper-could-put-patients-lives-at-risk #more-125745 ,