PRJNA637909: A gateway for SARS-CoV-2 infection in the human olfactory neuroepithelium
概要: Various reports indicate an association between COVID-19 and anosmia, suggesting an infection of the olfactory sensory epithelium, and thus a possible direct virus access to the brain. To test this hypothesis, we generated RNA-seq libraries from human olfactory neuroepithelia, in which we found substantial expression of the genes coding for the virus receptor angiotensin-converting enzyme-2 (ACE2), and for the virus internalization enhancer TMPRSS2. We analyzed a human olfactory single-cell RNA-seq dataset and determined that sustentacular cells, which maintain the integrity of olfactory sensory neurons, express ACE2 and TMPRSS2. We then observed that the ACE2 protein was highly expressed in a subset of sustentacular cells in human and mouse olfactory tissues. Finally, we found ACE2 transcripts in specific brain cell types, both in mice and humans. Sustentacular cells thus represent a potential entry door for SARS-CoV-2 in a neuronal sensory system that is in direct connection with the brain
项目整体设计: With the aim of exploring the potential expression of ACE2 and TMPRSS2 in olfactory sensory tissue, we collected biopsies via nasal endoscopic surgery from 4 adult patients. Samples of both nasal respiratory and olfactory sensory epithelia were harvested. Bulk tissue RNA was extracted, libraries generated, and sequenced
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Biopsies were snap frozen in liquid nitrogen immediately upon collection. |
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For RNA extraction, tissues were placed in RLT Buffer (Qiagen) with beta-mercaptoethanol and lysed with stainless steel balls (5mm diameter, Schieritz and Huenstein AG) using a homogenizer (Big Prep, MB Biomedicals). Total RNA was isolated using RNeasy Mini Kit (Qiagen) following the manufacturer protocol. The quality and quantity of total RNA were evaluated with a Bioanalyzer (Agilent). |
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Stranded cDNA libraries with suitable adapters for multiplexing were generated with Truseq RNA and DNA sample preparation kits (Illumina) following ribodepletion of the total RNA (200ng of total RNA per sample). Samples were multiplexed for sequencing in a HiSeq 2500 Sequencing system, generating 100bp single-end reads. |
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Illumina HiSeq 2500 |
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SARS-CoV-2 Receptors and Entry Genes Are Expressed in the Human Olfactory Neuroepithelium and Brain.
iScience . 2020-11-25 [PMID:
33251489]
