Gene Expression
Nebulas
Gene Expression NebulasSummary: Because epithelial surface pH affects airway inflammation and host defense, we study the benefit of an inhaled buffer in patients with asthma. Inhaled Optate is designed safely to raise airway surface pH and is well-tolerated in humans. Epithelial infection with SARS-CoV2 is attenuated by raising intracellular pH. Here, we show that Optate raises intracellular pH in primary normal human airway epithelial (NHAE) cells without affecting cell viability. Moreover, doses of Optate used in our human trials inhibit SARS-CoV-2 replication in NHAE cells. Specifically, Optate almost completely ablates viral plaque formation and nucleocapsid protein expression, decreasing expression of critical processing proteins. Optate could be a candidate to evaluate for treating COVID-19 patients.
Overall Design: Six samples of extracted and purified RNA from one line of primary human airway epithelial cells were processed for RNAseq analysis. This included three samples that were exposed to Optate for 48 hours and three control samples.
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| Growth Protocol: | 1 X 10^5 cells (second passage) were plated and allowed to adhere to the culture plate overnight. These cells underwent 48 hours of treatment (or control) and then underwent RNA extraction; both treatment and extraction are described below. |
| Treatment Protocol: | - |
| Extract Protocol: | Cells were lysed and RNA was extracted using the Qiagen RNeasy Plus Mini Kit per manufacturers protocol (Qiagen, USA, Cat No. 74134). |
| Library Construction Protocol: | - |
| Molecule Type: | poly(A)+ RNA |
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| Library Layout: | SINGLE |
| Library Strand: | Forward |
| Platform: | ILLUMINA |
| Instrument Model: | Illumina NextSeq 500 |
| Strand-Specific: | Specific |
| Data Resource | GEN Sample ID | GEN Dataset ID | Project ID | BioProject ID | Sample ID | Sample Name | BioSample ID | Sample Accession | Experiment Accession | Release Date | Submission Date | Update Date | Species | Race | Ethnicity | Age | Age Unit | Gender | Source Name | Tissue | Cell Type | Cell Subtype | Cell Line | Disease | Disease State | Development Stage | Mutation | Phenotype | Case Detail | Control Detail | Growth Protocol | Treatment Protocol | Extract Protocol | Library Construction Protocol | Molecule Type | Library Layout | Strand-Specific | Library Strand | Spike-In | Strategy | Platform | Instrument Model | Cell Number | Reads Number | Gbases | AvgSpotLen1 | AvgSpotLen2 | Uniq Mapping Rate | Multiple Mapping Rate | Coverage Rate |
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