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Principal Investigator: Susan Leilani Fink
Organization: UNIVERSITY OF WASHINGTON
Fiscal Year: 2019
Award: $233,250
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary
Zika virus (ZIKV) is a recently recognized global health threat due to the wide geographic distribution of risk
and potential for severe consequences of infection. There remains no antiviral in clinical use for ZIKV, or any
flavivirus, and many unanswered questions regarding the basic biology of these viruses. Our preliminary data
suggest that IRE1α, a component of the cellular response to endoplasmic reticulum (ER) stress, is required for
ZIKV replication. IRE1α also promotes optimal replication of flaviviruses related to ZIKV via an unknown
mechanism. Based on these data, we hypothesize that IRE1α inhibitors may be therapeutic against flaviviral
infection. This proposal aims to assess IRE1α activation in ZIKV infected cells, understand the molecular
mechanism(s) by which IRE1α supports ZIKV replication and determine whether targeting IRE1α could be
efficacious in a mouse model of infection.
IRE1α is activated during the unfolded protein response, a cellular pathway to detect and alleviate
dysfunctional protein folding in the ER. During ER stress, IRE1α initiates nonconventional splicing of XBP1
mRNA. Spliced XBP1 encodes a transcription factor, which upregulates targets involved in ER function.
IRE1α has more recently been shown to target other specific RNAs leading to their degradation. IRE1α
inhibitors are under evaluation for treatment of non-infectious human diseases, and we propose that this
project will provide preclinical evidence for the novel application of these drugs to treat flaviviral infection.
The experiments outlined in this proposal will extend our preliminary findings to important primary cell types
and currently circulating ZIKV strains (Aim 1). We will also determine whether the requirement for IRE1α in
ZIKV infection is XBP1-dependent or -independent, which will direct experiments to dissect the downstream
cellular processes required for ZIKV infection. In addition, we will identify the stage(s) of ZIKV replication
supported by IRE1α by systematically examining viral binding, RNA replication, protein synthesis and ER
remodeling (Aim 2). We predict that IRE1α most likely supports biogenesis of ER-derived viral replication
platforms, and will focus experiments on this hypothesis. Finally, we will determine whether IRE1α could be
targeted as an antiviral therapeutic strategy in mouse models of ZIKV infection (Aim 3). Together these aims
will reveal basic and practical insights into the biology of ZIKV replication, which are likely relevant for other
flaviviruses including established and potentially emergent human pathogens.
Terms: <Antiviral Agents><Antiviral Drugs><Antivirals><Basal Transcription Factor><Basal transcription factor genes><Binding><Biogenesis><Biology><Birth Defects><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Cell Body><Cell Function><Cell Line><Cell Process><Cell physiology><CellLine><Cells><Cellular Function><Cellular Physiology><Cellular Process><Clinical><Congenital Abnormality><Congenital Anatomic Abnormality><Congenital Anatomical Abnormality><Congenital Defects><Congenital Deformity><Congenital Malformation><Critical Paths><Critical Pathways><Cultured Cells><Data><Dengue><Dengue Fever><Drugs><ER stress><Endoplasmic Reticulum><Ergastoplasm><Evaluation><Flavivirus><Future><General Transcription Factor Gene><General Transcription Factors><General Viruses><Genetic><Geographic Distribution><Group B Arbovirus><HCV infection><Hepatitis C><Hepatitis C virus infection><Hepatitis, Viral, Non-A, Non-B, Parenterally-Transmitted><Hepatitus C><In Vitro><Infection><Infection prevention><Japanese B Encephalitis Virus><Japanese encephalitis virus><Knock-out><Knockout><Medical><Medication><Messenger RNA><Modeling><Molecular><Molecular Interaction><Nerve Cells><Nerve Unit><Neural Cell><Neurocyte><Neurons><Non-Polyadenylated RNA><Origin of Life><Pathway interactions><Pharmaceutic Preparations><Pharmaceutical Preparations><Pre-Clinical Model><Preclinical Models><Prevent infection><Protein Biosynthesis><Proteins><Publishing><RNA><RNA Gene Products><RNA Splicing><RNA replication><Ribonucleic Acid><Ribosomal Peptide Biosynthesis><Ribosomal Protein Biosynthesis><Ribosomal Protein Synthesis><Risk><Role><Short interfering RNA><Small Interfering RNA><Splicing><Strains Cell Lines><Subcellular Process><Testing><Therapeutic><Transcription Factor Proto-Oncogene><Transcription factor genes><Translating><Viral><Virus><Virus Replication><XBP1><XBP1 gene><Yellow fever virus><ZIKA><ZIKV><ZIKV infection><Zika Virus><Zika vaccine><Zika virus infection><Zika virus vaccine><anti-viral agents><anti-viral drugs><anti-virals><base><breakbone fever><cell type><cultured cell line><drug development><drug/agent><endoplasmic reticulum stress><experiment><experimental research><experimental study><global health><hep C><hepatitis non A non B><human disease><human pathogen><in vivo><inhibitor><inhibitor/antagonist><insight><mRNA><mouse model><murine model><neuronal><non A, non B hepatitis><non-A, non-B hepatitis><novel><pathway><pre-clinical><preclinical><protein folding><protein synthesis><response><siRNA><social role><therapeutic target><transcription factor><viral multiplication><viral replication><virus multiplication><zika infection><zika viral infection><zikav>