Identification of new host proteins essential for the replication of rhinovirus

Abstract

Rhinovirus (RV) infection causes the common cold and is a major trigger for acute exacerbations of asthma and chronic obstructive pulmonary disease (COPD). However, conventional antiviral approaches focused on directly targeting RV proteins have not led to the development of approved antiviral drugs. An alternative strategy is to interfere with cellular proteins that are exploited by RV during viral infection, which is less likely to lead to the emergence of antiviral resistance. However, current understanding of the host-pathogen interactions of RV is limited.

During infection, RV non-structural proteins (NSPs) interact with cellular proteins to subvert host cells and facilitate viral replication. Some of these cellular proteins, like GBF1, PI4KIIIβ and SETD3, were previously identified and proven to be good antiviral targets. However, we hypothesised that additional NSP-interacting cellular targets exist which could be exploited for antiviral development. To identify such new targets, we infected H1-HeLa cells with RV-A16 and pulled-down NSPs (2C/2BC, 3D/3CD, 3A/3AB and 3C/3CD) and their binding partners by NSP-specific affinity purification. Analysis of the pulled-down proteins by quantitative mass spectrometry (MS) revealed a specific enrichment in over 160 cellular proteins, including GBF1, PI4KIIIB and SETD3. Interestingly, numerous cellular proteins that were not previously shown to be involved in RV replication were enriched in independent pull-down experiments targeting several NSPs. Using small molecule inhibitors and siRNA knockdown, we show that some of these NSP interactors represent novel host factors that are essential for RV replication. In turn, this highlights their potential as drug targets for the treatment of RV infections.

Thesis embargoed until 31 July 2027.

Date of Award	Jul 2022
Original language	English
Awarding Institution	Queen's University Belfast
Sponsors	Northern Ireland Department for the Economy, Asthma and Lung UK & Medical Research Foundation
Supervisor	Aurelie Mousnier (Supervisor) & Ultan Power (Supervisor)