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        <datestamp>2023-05-30</datestamp>
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          <dc:identifier>https://doi.org/10.17044/SCILIFELAB.14188403</dc:identifier>
          <dc:title xml:lang="en">A phenomics approach for antiviral drug discovery - Images, analysis pipelines and feature data</dc:title>
          <dc:creator>Jonne Rietdijk</dc:creator>
          <dc:creator>https://orcid.org/0000-0001-5744-3206</dc:creator>
          <dc:creator>Aleksandra Pettke</dc:creator>
          <dc:creator>Polina Georgieva</dc:creator>
          <dc:creator>Maris Lapins</dc:creator>
          <dc:creator>Ulrika Warpman Berglund</dc:creator>
          <dc:creator>Ola Spjuth</dc:creator>
          <dc:creator>Marjo-Riitta Puumalainen</dc:creator>
          <dc:creator>Jordi Carreras Puigvert</dc:creator>
          <dc:subject xml:lang="en">Medical and Health Sciences</dc:subject>
          <dc:subject xml:lang="sv">Medicin och hälsovetenskap</dc:subject>
          <dc:subject xml:lang="en">Biochemistry</dc:subject>
          <dc:subject xml:lang="sv">Biokemi</dc:subject>
          <dc:description xml:lang="en">Abstract:

The current COVID-19 pandemic has highlighted the need for new and fast methods to identify novel or repurposed therapeutic drugs. Here we present a method for untargeted phenotypic drug screening of virus-infected cells, combining Cell Painting with antibody-based detection of viral infection in a single assay. We designed an image analysis pipeline for segmentation and classification of virus-infected and non-infected cells, followed by extraction of morphological properties. We show that the methodology can successfully capture virus-induced phenotypic signatures of MRC-5 human lung fibroblasts infected with Human coronavirus 229E (CoV-229E). Moreover, we demonstrate that our method can be used in phenotypic drug screening using a panel of nine host- and virus-targeting antivirals. Treatment with effective  antiviral compounds reversed the morphological profile of the host cells towards a non-infected state. The method can be used in drug discovery for morphological profiling of novel antiviral compounds on both infected and non-infected cells. 

Screen description: 
The images are of MRC-5 human lung fibroblasts infected with Human coronavirus 229E (CoV-229E) and treated with a panel of nine host- and virus-targeting antivirals. Cells are labelled with five labels that characterise seven cellular components (from the "Cell Painting" assay) as well as with a Coronavirus pan monoclonal antibody combined with a secondary antibody. This experiment consists of 5 plates. Each plate has 60 wells, and 9 fields of view per well. Each field was imaged in five channels (detection wavelengths), and each channel is stored as a separate, grayscale image file in TIFF format.The channel names (w1-w5) correspond to the following stains: w1 = Hoechst 33342 (HOECHST); w2= Coronavirus pan Monoclonal Antibody (FIPV3-70) + Goat Anti-Mouse IgG H&amp;L secondary antibody (MITO); w3= Wheat Germ Agglutinin/Alexa Fluor 555 + Phalloidin/Alexa Fluor 568 (PHAandWGA); w4= SYTO 14 green (SYTO); w5= Concanavalin A/Alexa Fluor 488 (CONC)
Organization of files:
1) Raw image data:
- MRC5_HCoV229_Plate1.tar.gz 
- MRC5_HCoV229_Plate2.tar.gz - MRC5_Plate3.tar.gz - MRC5_Plate4.tar.gz - MRC5_HCoV229_Plate5.tar.gz 
2) Image analysis pipelines (CellProfiler 4.0.7):
Cell Profiler project with a subset of images to try out the analysis pipeline:- Example_PipelineAndData.tar.gz 
Quality control, illumination correction and feature extraction pipelines:- AnalysisPipelines.tar.gz

3) Extracted feature data:
- features_MRC5_HCoV229_Plate1.tar.gz
- features_MRC5_HCoV229_Plate2.tar.gz- features_MRC5_Plate3.tar.gz- features_MRC5_Plate4.tar.gz- features_MRC5_HCoV229_Plate5.tar.gz
Metadata:
The file “Metadata_MRC5_HCoV229E_plate1-5.csv“ contains the metadata in CSV format, with the following fields:
- Plate_id: corresponds to the experimental plate
- Well: well allocation in the 96-well plate
- virus: "virus +" when cells are exposed to virus, and "virus -' for non-infected controls- Compound: name of compound
- Dose [μM]: dose of compound
For full information, see the manuscript to which this data is linked.</dc:description>
          <dc:rights>https://creativecommons.org/licenses/by/4.0/</dc:rights>
          <dc:publisher xml:lang="en">Uppsala University</dc:publisher>
          <dc:publisher xml:lang="sv">Uppsala universitet</dc:publisher>
          <dc:date>2021-03-22T00:00:00Z</dc:date>
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