| Author: | Char, Ho Fai |
| Title: | Screening of host microRNAs and potential target genes in response to the expression of SARS-CoV-2 derived small RNAs |
| Advisors: | Huang, Chien-ling (HTI) |
| Degree: | M.Sc. |
| Year: | 2022 |
| Subject: | Small interfering RNA COVID-19 (Disease) Hong Kong Polytechnic University -- Dissertations |
| Department: | Department of Health Technology and Informatics |
| Pages: | xiii, 79 pages : color illustrations |
| Language: | English |
| Abstract: | Introduction: Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and it is continued to be the global health and economic threat. MicroRNA (miRNA) of host and viral origin are indicated in dysregulating host immune response and worsens the diseases pathogenesis. Differential expression of host miRNA is associated with acute respiratory distress synfrome (ARDS). Therefore, in this project, three host miRNAs (miR-145-5p, miR-424-5p and miR-675-5p) were selected for expression profiling by transfecting different v-miRNAs encoded by SARS-CoV-2 N gene (v-miRNA-N-28612, 29094 and 29443) into Calu-3 cells. It is hypothesized that SARS-CoV-2 derived v-miRNA-N transfection significantly decreases the expression level of the host miRNAs. It is expected that this project can facilitate the discovery of host miRNA as a biomarker for disease progression and as a therapeutic target for treatment in the future. Aims: 1. To establish the expression profile of host miRNAs in response to transfection of SARS-CoV-2 derived v-miRNA-Ns into Calu-3 cells. 2. To predict the potential genes associated with ARDS targeted by the host miRNAs. Methods: Three different SARS-CoV-2 derived v-miRNA-Ns (28612, 29094 and 29443) were transfected individually and together into Calu-3 cells. The host miRNAs (miR-145-5p, miR-424-5p and miR-675-5p) were then detected by RT-qPCR. Relative fold changes of different host miRNAs were calculated as 2−∆∆Ct and analyzed by one-way analysis of variance (ANOVA) among different treatment groups. Statistical significance was defined as P-value smaller than 0.05. The ARDS-associated genes (MAP3K1, EGLN1, ITGB8, NLRP3 and CASP1) were selected for target gene prediction by host miRNAs using miRNA databases. The genes identified by at least two databases would be putative genes targeted by host miRNAs. Results: miR-424-5p had a significantly decreased expression level in individually v-miRNA-N transfected cells. No significant difference was found in expression level of miR-145-5p and miR-675-5p between v-miRNA-Ns transfected cells and mock mimics transfected cells. miR-145-5p, miR-424-5p and miR-675-5p were upregulated in three v-miRNA-Ns co-transfected cells. In target gene prediction, MAP3K1 and ITGB8 were targeted by miR-145-5p. EGLN1 and ITGB8 were targeted by miR-424-5p and miR-675-5p, respectively. Conclusions: It is concluded that miR-424-5p expression is strongly associated with SARS-CoV-2 infection. miR-424-5p could be a useful biomarker to estimate disease progression and a potential therapeutic target to improve disease severity. Further studies on overall host miRNA expression level could be conducted in SARS-CoV-2 infected cells. This could establish a more comprehensive host miRNA expression profile during SARS-CoV-2 infection. |
| Rights: | All rights reserved |
| Access: | restricted access |
Files in This Item:
| File | Description | Size | Format | |
|---|---|---|---|---|
| 6533.pdf | For All Users (off-campus access for PolyU Staff & Students only) | 1.73 MB | Adobe PDF | View/Open |
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