Orthogonal integration of virus-host interactions to develop broad-spectrum antivirals

This study will establish animal models for antiviral development against picornaviruses which cause commonly encountered infections in human (fits in the research field marked in the Form Section 1, item #3-3). We are engaged in a long-term collaboration with Dr. Tsu-An Hsu and other scientists at the Institute of Biotechnology and Pharmaceutical Research at the NHRI to develop therapeutic agents against RNA virus infections. In the current project, we would like to focus on the universal cellular factors that associate with different viruses in the family Picornaviridae as drug targets, and develop broad-spectrum antivirals. Picornaviruses include a large and diverse array of viruses (over 200 serotypes). Some picornavirus infections cause severe complications, such as poliomyelitis, encephalitis, myocarditis, meningitis, neurological pulmonary edema, and even death. The vast number of serotypes in this virus family makes it challenging to develop vaccines that can prevent multiple picornaviruses. Therefore, broad-spectrum antivirals are warranted. There are four specific aims in the proposal, (1) to search for common host factors involved in the replication of different picornaviruses, (2) to integrate omics data and identify the mechanistic paradigm (linchpins) as targets for broad-spectrum antivirals, (3) to establish a high throughput screening assay and identify potential inhibitors, and (4) to evaluate the functionality of inhibitors in newly established animal models (new animal models for EV-A71, Coxackie A, Coxackie B, EV-D68, and Rhinoviruses). In Specific Aim 1, in the past one year, we have identified several host factors which are involved in viral replication of EV-D68 by using CRISPR-Cas9 screening, including EIF6, LAMA2, MRPS24, RPS11, KCTD11, PDXDC1, ST6GAL1, SLC35A1, SLC35A2, and SCARB2. In addition to the genes (ST6GAL1, SLC35A1, SLC35A2, and SCARB2) that have been reported to require for EV-D68 or EV-A71 infection, other top-ranking genes (EIF6, LAMA2, MRPS24, RPS11, KCTD11, and PDXDC1) will be used for further validation and to investigate their biological roles in virus replication. In addition, metabolic labeling method combined with the advanced proteomic analysis and NGS will be also applied to explore the mechanisms of these novel factors involved in picornavirus infection. Dr. Hsu’s team (NHRI) will be actively involved in establishing the high throughput screening assay based on the function of the cell targets, for example, interference assay for SCARB2, a receptor for several enteroviruses. Once the high throughput screening assays are established, the compound library collected by the Institute of Biotechnology and Pharmaceutical Research at the NHRI will be applied toward an antiviral drug-screening assay. We believe this project will not only contribute to broad-spectrum antiviral development, but also provide insight into the complex molecular interactions between viruses and hosts that is crucial to understand their impact on viral pathogenesis.

Project ID:PG10801-0151
External Project ID:NHRI-EX108-10833SI