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Transcriptome analysis of human colonic organoids infected SARS-CoV-2

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE179949 | GEO | 2022/02/16

REPOSITORIES: GEO

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Project description:Coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is a global public health emergency. COVID-19 typically manifests as a respiratory illness but an increasing number of clinical reports describe gastrointestinal (GI) symptoms. This is particularly true in children in whom GI symptoms are frequent and viral shedding outlasts viral clearance from the respiratory system. By contrast, fetuses seem to be rarely affected by COVID-19, although the virus has been detected in placentas of affected women. These observations raise the question of whether the virus can infect and replicate within the stomach once ingested. Moreover, it is not yet clear whether active replication of SARS-CoV-2 is possible in the stomach of children or in fetuses at different developmental stages. Here we show the novel derivation of fetal gastric organoids from 8-21 post-conception week (PCW) fetuses, and from pediatric biopsies, to be used as an in vitro model for SARS-CoV-2 gastric infection. Gastric organoids recapitulate human stomach with linear increase of gastric mucin 5AC along developmental stages, and expression of gastric markers pepsinogen, somatostatin, gastrin and chromogranin A. In order to investigate SARS-CoV-2 infection with minimal perturbation and under steady-state conditions, we induced a reversed polarity in the gastric organoids (RP-GOs) in suspension. In this condition of exposed apical polarity, the virus can easily access viral receptor angiotensin-converting enzyme 2 (ACE2). The pediatric RP-GOs are fully susceptible to infection with SARS-CoV-2, where viral nucleoprotein is expressed in cells undergoing programmed cell death, while the efficiency of infection is significantly lower in fetal organoids. The RP-GOs derived from pediatric patients show sustained robust viral replication of SARS-CoV-2, compared with organoids derived from fetal stomachs. Transcriptomic analysis shows a moderate innate antiviral response and the lack of differentially expressed genes belonging to the interferon family. Collectively, we established the first expandable human gastric organoid culture across fetal developmental stages, and we support the hypothesis that fetal tissue seems to be less susceptible to SARS-CoV-2 infection, especially in early stages of development. However, the virus can efficiently infect gastric epithelium in pediatric patients, suggesting that the stomach might have an active role in fecal-oral transmission of SARS-CoV-2.

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Transcriptome analysis of human colonic organoids infected SARS-CoV-2

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