Project description:RNAseq analysis of human lung AT2 cell organoids infected with CoV-2, treated with inhibitors Results: Differentially expressed genes after infection and treatment with inhibitors.
Project description:Purpose: To identify the diferentially expressed genes in SARS-CoV-2 susceptible and resistant organoids during the ifnection. Method: We selected 3 susceptible (C8, C9, and C10)- and 3 restant (C1, C2, and C7)-organoids lines and infected SARS-CoV-2 at multiplicity of infection (MOI) of 4 for 24 and 72 hrs. The RNAs were collected and then sequenced by CEL-seq2. Sequencing was performed on Illumina NovaSeq 6000. Results: Longitudinal transcriptome analyses identified robust yet late transcriptional changes induced by SARS-CoV-2, the magnitude of which corresponded to the levels of viral infection.
Project description:The distal lung contains terminal bronchioles and alveoli that facilitate gas exchange. Three-dimensional in vitro human distal lung culture systems would strongly facilitate investigation of pathologies including interstitial lung disease, cancer, and SARS-CoV-2-associated COVID-19 pneumonia. We generated long-term feeder-free, chemically-defined culture of distal lung progenitors as organoids derived from single adult human alveolar epithelial type II (AT2) or KRT5+ basal cells. AT2 organoids exhibited AT1 transdifferentiation potential while basal cell organoids developed lumens lined by differentiated club and ciliated cells. Single cell analysis of basal organoid KRT5+ cells revealed a distinct ITGA6+ITGB4+ mitotic population whose proliferation further segregated to a TNFRSF12Ahi subfraction comprising ~10% of KRT5+ basal cells, residing in clusters within terminal bronchioles and exhibiting enriched clonogenic organoid growth activity. Distal lung organoids were created with apical-out polarity to display ACE2 on the exposed external surface, facilitating SARS-CoV-2 infection of AT2 and basal cultures and identifying club cells as a novel target population. This long-term, feeder-free organoid culture of human distal lung, coupled with single cell analysis, identifies unsuspected basal cell functional heterogeneity and establishes a facile in vitro organoid model for human distal lung infections including COVID-19-associated pneumonia.
Project description:We performed RNA-Seq of SARS-Cov-2 infection in human bronchial epithelium organoids. The organoids were infected with SARS-Cov-2 for 48hours or 72hours respectively, and compared with uninfected mock control.
Project description:We performed RNA-Seq of SARS-Cov-2 infection in human airway epithelium organoids. The organoids were infected with SARS-Cov-2 for 24hours or 48hours respectively, and compared with uninfected mock control.
Project description:Purpose: The goal of this study is to understand the response and pathology of the epigenome upon infection with SARS-CoV-2 in lung and heart tissues Methods: AT2 and induced cardiomyocytes were infected (MOI 4) with SARS-CoV-2 or Mock infection control for 48 hours followed by chromatin immunoprecipitation and sequencing. Results: global differences in hPTMs were observed upon infection related to SARS-CoV-2 histone mimicry Conclusions: SARS-CoV-2 sucks
Project description:Alveolar type 2 (AT2) cells function as stem cells in the adult lung and aid in injury-repair. The current study aimed to understand the signaling events that control differentiation of this therapeutically relevant cell type during human development through differentiation of lung progenitor organoids to AT2 cells and benchmarking against primary AT2 organoids.
Project description:The SARS-CoV-2 virus has already caused over a million COVID-19 cases and over fifty-thousand deaths globally. There is an urgent need to create novel models to study SARS-CoV-2 virus using human disease-relevant cells and tissues to understand key features of virus biology. We present a platform comprised of nine different cell and organoid derivatives from human pluripotent stem cells (hPSCs) representing all three primary germ layers, including lung progenitors and alveolar type II (AT2) cells, pancreatic endocrine cells, liver organoids, endothelial cells, cardiomyocytes, macrophages, microglia, and both cortical and dopaminergic neurons. We systematically probed which cell types are permissive to SARS-CoV-2 infection. Human pancreatic beta cells and hepatocytes were strikingly permissive to SARS-CoV-2 infection, further validated using adult primary human islets and liver organoids. Both in vitro and in a humanized mouse model, human lung progenitors and AT2 cells express the ACE2 viral receptor and were highly permissive to SARS-CoV-2 infection. Transcriptomic analysis following SARS-CoV-2 infection of hPSC-derived pancreatic and lung organoids revealed upregulation of chemokines but not type I/III interferon signaling, similar to what was seen in primary human COVID-19 pulmonary infection. Therefore, hPSC-derived cells phenocopy human COVID-19 disease and provide a valuable resource to understand SARS-CoV-2 biology and search for novel therapeutics.