Identification of cell lines CL-14, CL-40 and CAL-51 as suitable models for SARS-CoV-2 infection studies
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
The SARS-CoV-2 pandemic is a major global threat that sparked global research efforts. Pre-clinical and biochemical SARS-CoV-2 studies firstly rely on cell culture experiments where the importance of choosing an appropriate cell culture model is often underestimated. We here present a bottom-up approach to identify suitable permissive cancer cell lines for drug screening and virus research. Human cancer cell lines were screened for the SARS-CoV-2 cellular entry factors ACE2 and TMPRSS2 based on RNA-seq data of the Cancer Cell Line Encyclopedia (CCLE). However, experimentally testing permissiveness towards SARS-CoV-2 infection, we found limited correlation between receptor expression and permissiveness. This underlines that permissiveness of cells towards viral infection is determined not only by the presence of entry receptors but is defined by the availability of cellular resources, intrinsic immunity, and apoptosis. Aside from established cell culture infection models CACO-2 and CALU-3, three highly permissive human cell lines, colon cancer cell lines CL-14 and CL-40 and the breast cancer cell line CAL-51 and several low permissive cell lines were identified. Cell lines were characterised in more detail offering a broader choice of non-overexpression in vitro infection models to the scientific community. For some cell lines a truncated ACE2 mRNA and missense variants in TMPRSS2 might hint at disturbed host susceptibility towards viral entry.
Article activity feed
-
-
SciScore for 10.1101/2020.07.09.195040: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Western blots: Antibodies against ACE2 (ab15348), TMPRSS2 (ab109131), and GAPDH (ab8245) were obtained from Abcam (Cambridge, UK). ACE2suggested: NoneTMPRSS2suggested: (Abcam Cat# ab109131, RRID:AB_10863728)GAPDHsuggested: (Abcam Cat# ab8245, RRID:AB_2107448)Primary antibody used to stain dsRNA was mouse anti-dsRNA, clone rJ2 (Merck Millipore, Cat. no. MABE1134); secondary antibody was goat anti-rabbit-Cy5 (Life Technologies, Cat. no. A10523). anti-dsRNAsugg…SciScore for 10.1101/2020.07.09.195040: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Western blots: Antibodies against ACE2 (ab15348), TMPRSS2 (ab109131), and GAPDH (ab8245) were obtained from Abcam (Cambridge, UK). ACE2suggested: NoneTMPRSS2suggested: (Abcam Cat# ab109131, RRID:AB_10863728)GAPDHsuggested: (Abcam Cat# ab8245, RRID:AB_2107448)Primary antibody used to stain dsRNA was mouse anti-dsRNA, clone rJ2 (Merck Millipore, Cat. no. MABE1134); secondary antibody was goat anti-rabbit-Cy5 (Life Technologies, Cat. no. A10523). anti-dsRNAsuggested: (Millipore Cat# MABE1134, RRID:AB_2819101)anti-rabbit-Cy5suggested: NoneExperimental Models: Cell Lines Sentences Resources Supernatants of infected cells were taken 5 dpi, serially diluted, and used to infect confluent Vero E6 cells on 96-well cell culture plates for one hour. Vero E6suggested: NoneSoftware and Algorithms Sentences Resources Briefly, alignment files were downloaded from CGHub via genetorrent, sorted (samtools 0.1.19) and converted to fastq files (bedtools v2.21.0). samtoolssuggested: (SAMTOOLS, RRID:SCR_002105)bedtoolssuggested: (BEDTools, RRID:SCR_006646)STAR (2.5.3a) served as read mapper to the human genome Gencode 26, HTSeq (0.11.3) was used as count tool. STARsuggested: (STAR, RRID:SCR_015899)HTSeqsuggested: (HTSeq, RRID:SCR_005514)Raw counts were normalised and transformed to FPKM (fragments per kilobase million) via R/Bioconductor (3.6.9) loading DESeq2 (1.26.0) and visualised via ggplot2 (3.1.1). DESeq2suggested: (DESeq, RRID:SCR_000154)ggplot2suggested: (ggplot2, RRID:SCR_014601)Proteins on nitrocellulose membranes were visualized with the biotin/streptavidin-horseradish peroxidase system (GE Healthcare; Little Chalfont, UK) in combination with the “Renaissance Western Blot Chemoluminescence Reagent” (Perkin Elmer; Waltham, MA, USA). GE Healthcaresuggested: (GE Healthcare, RRID:SCR_000004)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: 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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- No conflict of interest statement was detected. If there are no conflicts, we encourage authors to explicit state so.
- 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.
-
