Abstract
Dengue virus (DENV) is the cause of the most important arthropod-borne viral disease in the world. Furthermore, its range is spreading partly as a result of climate change and population movement. Knowledge of dengue pathogenesis is incomplete. The only licensed vaccine (Dengvaxia®, CYD-TDV) is not fully effective, and no clinically approved antiviral drugs to treat DENV infections are available at present. In this study, the proteomes of DENV-2 infected Huh-7 cells, replicon containing Huh-7 cells and the sera of DEN patients were systematically analyzed using quantitative mass spectrometry with the aim of increasing the understanding of DENV pathogenesis. Further aims were to identify potential biomarkers for DEN severity and identify degradation pathways for key host proteins during DENV infection.Bioinformatic analysis revealed that the main effect of DENV infection was a significant decrease in the abundance of proteins involved in complement and coagulation cascade and lipid metabolic processes. Evidence is presented in this study that DENV induces host cell arrest in the G1 phase. An explanation has been proposed for the unexpected decreased levels of cellular positive liver acute phase proteins in DENV-2 infected Huh-7 cells. Also, an explanation for the decreased levels of VTN in DENV-2 infected Huh-7 cells and the sera of DEN patients has been proposed.
The results confirm that replicon containing Huh-7 cells are a good surrogate for DENV-2 infected Huh-7 cells for in vitro studies of DENV pathogenesis. The results demonstrate the power of DENV infected Huh-7 / replicon cell systems to analyze viral replication. A model has been proposed to explain how DENV replicons are able to carry out stable, robust replication in Huh-7 cells. An analysis of the biosafety of replicons as a platform for viral studies has been carried out in this study.
Several host proteins that are targeted for degradation by the UPS were identified including DNMT1,UHRF1, PCNA, RAD51, POL2, CDK2, CDK4 and ELL2. Several host proteins that are targeted by DENV for degradation by the ALP were identified including SERPINA1, CEPBA, COCH, SCD, HMGCR, MSOM1 and Reelin. Several potential potent host antiviral proteins to treat DEN patients have been identified including CEPBA, COCH and Reelin. The results indicate that the identified host antiviral proteins are broad spectrum. Two treatment plans to treat DEN patients have been proposed and a public health program targeted at high nutritional status children has been proposed. The results of the analysis suggest that DENV-2 infected Huh-7 cells are a good model for in vivo DEN studies using clinical sera, especially those relating to coagulation and complement cascades, lipid metabolism and liver acute phase proteins.
Evidence is presented in this study that several host proteins are potential biomarkers to discriminate between severe dengue and lesser grades of DENV infection including ANXA3, OMD, IGFB2 and LCP1. Evidence is presented in this study that several host proteins are potential biomarkers that discriminate between dengue infection and pyrexia of unknown origin. Evidence is presented in this study that several host proteins are potential biomarkers to discriminate between severe dengue and pyrexia of unknown origin including OMD, IGFB2 and LCP1.
| Date of Award | 3 Oct 2023 |
|---|---|
| Original language | English |
| Awarding Institution |
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| Supervisor | Andrew D Davidson (Supervisor) |
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