Preliminary analysis of scRNA sequencing of children’s lung tissues excludes the expression of SARS-CoV-2 entry related genes as the key reason for the milder syndromes of COVID-19 in children
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Abstract
To explore whether the expression levels of viral-entry associated genes might contribute to the milder symptoms in children, we analyzed the expression of these genes in both children and adults’ lung tissues by single cell RNA sequencing (scRNA-seq) and immunohistochemistry (IHC). Both scRNA-seq and IHC analyses showed comparable levels of the key genes for SARS-CoV-2 entry in children and adults, including ACE2, TMPRSS2 and FURIN , suggesting that instead of lower virus intrusion rate, other factors are more likely to be the key reasons for the milder symptoms of SARS-CoV-2 infected children.
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SciScore for 10.1101/2020.05.25.20110890: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: The study was approved by the Institutional Review Board and the Ethics Committee of Shanghai Children’s Medical Center (SCMCIRB-K2020034), and written informed consent was obtained from (the parents of) each patient.
Consent: The study was approved by the Institutional Review Board and the Ethics Committee of Shanghai Children’s Medical Center (SCMCIRB-K2020034), and written informed consent was obtained from (the parents of) each patient.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources After deparaffinization, antigen retrieval and blocking, … SciScore for 10.1101/2020.05.25.20110890: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: The study was approved by the Institutional Review Board and the Ethics Committee of Shanghai Children’s Medical Center (SCMCIRB-K2020034), and written informed consent was obtained from (the parents of) each patient.
Consent: The study was approved by the Institutional Review Board and the Ethics Committee of Shanghai Children’s Medical Center (SCMCIRB-K2020034), and written informed consent was obtained from (the parents of) each patient.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources After deparaffinization, antigen retrieval and blocking, the slides were stained using the Leica Bond (Leica Biosystems) automated staining system with the following primary antibodies for 60 min at room temperature: anti-ACE2 antibody (rabbit, ab108252 from Abcam, 1:100) and anti-TMPRSS2 antibody (rabbit, ab92323 from Abcam, 1:2000) for cohort 1; anti-ACE2 antibody (mouse, 66699-1-Ig, Proteintech, 1:1200) and anti-FURIN antibody (rabbit, 18413-1-AP, Proteintech, 1:1200) for cohort 2. anti-ACE2suggested: (Enzo Life Sciences Cat# ALX-804-722-C100, RRID:AB_11180102)anti-TMPRSS2suggested: (Abcam Cat# ab92323, RRID:AB_10585592)anti-FURINsuggested: (Proteintech Cat# 18413-1-AP, RRID:AB_2878542)After three washes, the slides were then incubated with HRP-conjugated goat anti-rabbit antibody (cohort 1) or biotin-conjugated anti-mouse/rabbit secondary antibody (cohort 2) for 15-30 min at room temperature. anti-rabbitsuggested: Noneanti-mouse/rabbitsuggested: (Biorbyt Cat# orb27530, RRID:AB_10954533)Software and Algorithms Sentences Resources Briefly, FastQC (V0.11.7), fastp (1) FastQCsuggested: (FastQC, RRID:SCR_014583)STAR aligner (2.5.3a) and featureCounts (1.6.2) were used for quality evaluation, trimming, alignment and transcript counting, respectively STARsuggested: (STAR, RRID:SCR_015899)featureCountssuggested: (featureCounts, RRID:SCR_012919)Adapters and poly(A) tails were trimmed before aligning read 2 to GRCh38 with Ensembl 92 annotation. Ensemblsuggested: (Ensembl, RRID:SCR_002344)Integrated analysis of children lung data with public adult lung data: The public data of 8 healthy adults were downloaded from Gene Expression Omnibus (GSE122960) Gene Expression Omnibussuggested: (Gene Expression Omnibus (GEO, RRID:SCR_005012)Statistical analyses of IHC scores were performed in GraphPad Prism 8 using Mann-Whitney test with an exact p value computed. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: Thank you for sharing your data.
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.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: Please consider improving the rainbow (“jet”) colormap(s) used on page 13. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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