SARS-CoV-2 Cell Entry Factors ACE2 and TMPRSS2 are Expressed in the Pancreas but are Not Enriched in Islet Endocrine Cells
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Abstract
Reports of new-onset diabetes and diabetic ketoacidosis in individuals with COVID-19 have led to the hypothesis that SARS-CoV-2, the virus that causes COVID-19, is directly cytotoxic to pancreatic islet β cells. This would require binding and entry of SARS-CoV-2 into host β cells via cell surface co-expression of ACE2 and TMPRSS2, the putative receptor and effector protease, respectively. To define ACE2 and TMPRSS2 expression in the human pancreas, we examined six transcriptional datasets from primary human islet cells and assessed protein expression by immunofluorescence in pancreata from donors with and without diabetes. ACE2 and TMPRSS2 transcripts were low or undetectable in pancreatic islet endocrine cells as determined by bulk or single cell RNA sequencing, and neither protein was detected in α or β cells from these donors. Instead, ACE2 protein was expressed in the islet and exocrine tissue microvasculature and also found in a subset of pancreatic ducts, whereas TMPRSS2 protein was restricted to ductal cells. The absence of significant ACE2 and TMPRSS2 co-expression in islet endocrine cells reduces the likelihood that SARS-CoV-2 directly infects pancreatic islet β cells through these cell entry proteins.
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SciScore for 10.1101/2020.08.31.275719: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: The Vanderbilt University Institutional Review Board has declared that studies on de-identified human pancreatic specimens do not quality as human subjects research. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Peptide Competition: We combined 1μg of anti-ACE2 antibody (ab15348) with or without 10μg of the immunizing human ACE2 peptide (ab15352) in antibody buffer solution (0.1% Triton X-100/1% BSA/1X PBS) and incubated overnight at 4°C with gentle agitation. anti-ACE2suggested: (Abcam Cat# ab15348, RRID:AB_301861)Software and Algorithms Sentences …SciScore for 10.1101/2020.08.31.275719: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: The Vanderbilt University Institutional Review Board has declared that studies on de-identified human pancreatic specimens do not quality as human subjects research. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Peptide Competition: We combined 1μg of anti-ACE2 antibody (ab15348) with or without 10μg of the immunizing human ACE2 peptide (ab15352) in antibody buffer solution (0.1% Triton X-100/1% BSA/1X PBS) and incubated overnight at 4°C with gentle agitation. anti-ACE2suggested: (Abcam Cat# ab15348, RRID:AB_301861)Software and Algorithms Sentences Resources GraphPad Prism v8 was used to generate plots in Figure 1A. Single Cell RNA-seq Data Acquisition: GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Raw gene count matrices were extracted from existing single cell RNA-seq datasets (Baron et al., 2016; Segerstolpe et al., 2016; Yang et al., 2020) and from the Human Pancreas Analysis Program (HPAP) Database (https://hpap.pmacs.upenn.edu), a Human Islet Research Network consortium. Human Pancreas Analysis Programsuggested: (HIRN Human Pancreas Analysis Program, RRID:SCR_016202)Gene count matrices were further analyzed using the R package Seurat version 3.1 (Stuart et al., 2019) Seuratsuggested: (SEURAT, RRID:SCR_007322)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:As outlined below in the “Limitations of the Study” section, additional studies are needed to investigate whether direct SARS-CoV-2 infection of β cells occurs or is detrimental to β cell health or function by other mechanisms. However, based on current data, it appears that the interaction of diabetes and SARS-CoV-2 is mediated by systemic inflammation and/or metabolic changes in other organs such as liver, muscle or adipose tissue. Limitations of the Study: We did not directly measure SARS-CoV-2 binding or entry into human β cells but instead assessed expression of canonical, co-receptors, ACE2 and TMPRSS2, in the human pancreas. While studies culturing SARS-CoV-2 and human islets in vitro will be of considerable interest, such experiments may be challenging to interpret as the islet isolation process itself may impact ACE2 or TMPRSS2 expression or susceptibility to viral infection. In addition, infection of islet cells within an intact, isolated islet with a variety of viruses has proven difficult as is selection of an appropriate ratio of viral particles and islet cells reflective of the in vivo environment. Furthermore, it is not clear if infection of an isolated islet accurately models the physiology of a vascularized, innervated islet within the context of the pancreas. While the preponderance of research suggests that ACE2 and TMPRSS2 are the primary means for SARS-CoV-2 entry into host cells, knowledge about additional pathways and/or mechanisms of SARS-CoV-2 cell en...
Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
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- No protocol registration statement was detected.
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