Next Generation Sequencing of SARS-CoV-2 infected lung epithelial cells derived from iPSC
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ABSTRACT:
PROVIDER: PRJNA786238 | ENA |
REPOSITORIES: ENA
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Project description:We present evidence on how a lung epithelial cell culture system based on human induced pluripotent stem cells (hiPSC) could emerge as a relevant and sensitive platform for modelling SARS-CoV-2 infection. In order to develop a human model system, we differentiated hiPSC into lung epithelial lineage (airway and alveolar cells) through a three-step protocol. De novo generated cells were authenticated by spatiotemporal expression of stage specific markers including SOX2, FOXJ1 and CC10 in proximal cells and SOX9, FOXP2, SFTPC, MUC5AC, and CC10 in distal cells. Further we showed upregulation of ACE2 and TMPRSS2, the two proteins responsible for SARS-CoV-2 binding and invasion into the host lung in our iPSC-derived proximal and distal lung epithelial cells. In order to find out whether our in-house created lung cell culture system was permissive to SARS-CoV-2 infection, two viral doses representing two multiplicities of infections (MOIs) was tested. Plaque assay, RT-PCR, and ICC results demonstrated that the virus was able to enter and replicate inside these cells in a reproducible manner. Further, transcriptomics (RNA seq) data uncovered a robust epithelial cell-specific response to virus infection including perturbation of metabolic processes, disruption in alveolar and airway maturation program, in addition to the well-known upregulation of antiviral defense and immune responses.
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Project description:To investigate the impact of SARS-CoV-2 on the peripheral nervous system, we generated human sensory neurons from induced pluripotent stem cells (iPSC) that we further infected with the SARS-CoV-2 strain WA1/2020 for 48 hours.
2023-08-04 | GSE227529 | GEO
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2023-08-04 | GSE227528 | GEO
Project description:HAE cultures were infected with SARS-CoV, SARS-dORF6 or SARS-BatSRBD and were directly compared to A/CA/04/2009 H1N1 influenza-infected cultures. Cell samples were collected at various hours post-infection for analysis. Time Points = 0, 12, 24, 36, 48, 60, 72, 84 and 96 hrs post-infection for SARS-CoV, SARS-dORF6 and SARS-BatSRBD. Time Points = 0, 6, 12, 18, 24, 36 and 48 hrs post-infection for H1N1. Done in triplicate for RNA Triplicates are defined as 3 different wells, plated at the same time and using the same cell stock for all replicates. Time matched mocks done in triplicate from same cell stock as rest of samples. Culture medium (the same as what the virus stock is in) will be used for the mock infections. Infection was done at an MOI of 2 for SARS viruses and an MOI of 1 for H1N1.
2013-07-31 | E-GEOD-47962 | biostudies-arrayexpress
Project description:The coronavirus disease 2019 (COVID-19) pandemic has affected nearly 700 million and claimed more than 6 million lives. However, the large heterogeneity in disease susceptibility and severity of illness (SOI) remains poorly understood. A growing body of evidence suggests that alveolar epithelial cell type 2 (AT2), which constitute 10-15% of all alveolar cells and are critical for alveolar homeostasis, are the primary targets of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection and damage to AT2s may directly contribute to disease severity and poor prognosis in COVID-19 patients. Our in-vitro modeling of SARS-CoV-2 infection, in well-characterized, highly uniform, (r2 at 95% CI = 0.92 ± 0.03), induced pluripotent stem cell (iPSC) derived AT2s from 10 participants of our Mexican American Family Study, showed interindividual variability in infection susceptibility and the post-infection cellular viral load. To understand the underlying mechanism of the AT2’s capacity to regulate SARS-CoV-2 infection and cellular viral load, we performed a systematic characterization of the pre- and post-SARS-CoV-2 challenged AT2 transcriptomes. A total of 1,393 genes were found significantly (Oneway ANOVA FDR corrected p ≤ 0.05; FC abs ≥ 2.0) differentially expressed (DE) between pre- and post-SARS-CoV-2 infection-challenged AT2 and suggest significant upregulation of viral infection-related cellular innate immune response pathways (p-value ≤ 0.05; activation z-score ≥ 3.5), whilst the cholesterol and xenobiotic related metabolic pathways were significantly downregulated (p-value ≤ 0.05; activation z-score ≤ - 3.5). Interestingly, pre-infection expression of 238 DE genes showed a high correlation with the post-infection SARS-CoV-2 viral load (FDR corrected p-value ≤ 0.05, and r2-absolute ≥ 0.57). The 85 genes whose expression was negatively correlated with the viral load showed significant enrichment in viral recognition and cytokine-mediated innate immune GO biological processes (p-value range: 4.65x10-10 to 2.24x10-6). The 153 genes whose expression was positively correlated with the viral load showed significant enrichment in cholesterol homeostasis, extracellular matrix, and MAPK/ERK pathway-related GO biological processes (p-value range: 5.06x10-5 to 6.53x10-4). Overall, our results strongly suggest that AT2s pre-infection innate immunity and metabolic state affects their susceptibility to SARS-CoV-2 infection and viral load.
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| 12454 | ecrin-mdr-crc