Although Zika virus (ZIKV) was discovered almost 70 years ago in Zika forest in Uganda, it is only recently recognized as a global public threat. Historically, ZIKV infection has been associated with asymptomatic illnesses such as rash, arthralgia, and conjunctivitis. However, recently several studies have found that ZIKV infection causes severe fetal abnormalities that include microcephaly in infants and the neurological disorder GBS in adults. To date, there is no vaccine or antiviral therapy, and limited information is available for accurately predicting the outcome as well as the resulting complications due to ZIKV infection. There is a knowledge gap in understanding the basic pathogenesis of this virus, which is critical for developing strategies to combat this disease. The role of host factors especially that of host lipids in ZIKV pathogenesis is largely unknown. The profound success of flaviviruses such as ZIKV in causing disease depends on its ability to successfully utilize the host's cellular machineries including the lipid biosynthesis pathway to subvert the immune system. Our understanding of these processes especially that of the host lipid biology in the context of ZIKV is limited.
The aim of the proposed work is to study the role of host's lipid biosynthesis in the entry, replication, assembly and release of ZIKV. For this purpose, individual knockout Huh7 and SH-SY5Y cell lines that lack key genes involved in the biosynthesis of lipids will be generated using CRISPR/Cas9-technology. To complement our genetic approach, small molecule inhibitors will be used to block these key pathways, and their impact in ZIKV infection will be investigated. In addition to plaque assay, automated high-content microscopy will be performed in ZIKV infected mutant and control cells to define the function of host lipids at different stages of ZIKV infection. One of the major cellular antimicrobial processes, autophagy, will be assessed in both knockout and control cells during ZIKV infection. Using lipid reporter tools we also plan to investigate the spatial and temporal (re-)distribution of lipids during ZIKV infection. Understanding these pathways or processes essential for the life cycle of ZIKV is crucial, as they represent potential targets for new therapeutics.

Public Health Relevance

Although Zika virus has recently emerged as a significant global health threat, there is no vaccine or antiviral therapy, and limited information is available for accurately predicting the outcome of ZIKV infection as well as resulting complications. Understanding the pathogenesis of this virus and the host factors including cellular lipids critical for entry, replication and release of the virus is crucial for developing strategies to combat this disease. We will therefore aim to identify and characterize the host lipid factors that are used by Zika virus during infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI133631-01
Application #
9387983
Study Section
Special Emphasis Panel (ZAI1)
Program Officer
Challberg, Mark D
Project Start
2017-09-01
Project End
2019-08-31
Budget Start
2017-09-01
Budget End
2018-08-31
Support Year
1
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Oregon Health and Science University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239