Flaviviruses need and manipulate host cell pathways at every step of their infectious cycle. Since many host cell pathways are controlled by protein kinases, this class of host proteins is intimately involved in modulating flaviviral replication both negatively, ie. by controlling innate immune responses, as well as positively, by inducing a virus-supportive environment. The goal of this sub-project is to identify those kinase-controlled host cell pathways that support the flavivirus life cycle. Our preliminary data show that broad-spectrum protein tyrosine kinase inhibitors prevent flaviviral entry whereas MAP-kinase inhibitors interfere with replication. Moreover, we previously demonstrated that Src-kinase inhibitors block flaviviral egress due to a requirement of the c-yes kinase for this step. To identify both specific inhibitors and specific kinases controlling each step of the viral life cycle we will use a chemical genomics approach. We will employ cellbased, high content screening of kinase-directed small molecule libraries to identify kinase inhibitors interfering with different steps of the Dengue-virus (DENV) life cycle. In secondary screens we will select for compounds with a high selectivity index, broad cell type specificity and activity against other flaviviruses (WNV, JEV, YFV). Active compounds will likely inhibit several related kinases thus minimizing the problem of redundancy of signaling pathways and tissue specific expression. The kinases targeted by a given compound will be identified by combining computational, functional genomics and biochemical approaches. Based on these results we will generate models of kinase networks controlling flavivirus infection and examine thise models experimentally by phosphoproteomics and by targeted gene knockdown. Finally, we will examine the therapeutic potential of kinase-targeted antivirally active compounds by examining selected compounds in vivo using a murine model West-Nile Virus infection. The efficacy, toxicity and bio-availability of kinase-directed compounds will be compared with the in vivo activity of TYT-1 analogs (sultam thioureas) generated in sub-project 3 of this program. At the end of these comprehensive studies, we anticipate to have gained further insights into the interaction of flaviviruses with their host cell, identified novel targets for antviral drugs discovery, and characterized novel small molecule inhibitors that are active against flaviviruses in vitro and in vivo.

Public Health Relevance

There is a need for specific therapies for flaviviruses such as Dengue, West Nile, Japanese Encephalitis and Yellow fever virus, but their small genome size limits the number of enzymes for antiviral drug discovery. In contrast, the host expresses hundreds of proteins needed for the virus to replicate, some of them known drug targets. We propose to systematically study how host cell kinases, one of the most intensely studied class of drug targets, control infection by flaviviruses by screening for antiviral activity of kinase inhibitors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54AI081680-05
Application #
8436325
Study Section
Special Emphasis Panel (ZAI1-DDS-M)
Project Start
Project End
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
5
Fiscal Year
2013
Total Cost
$555,720
Indirect Cost
$96,063
Name
Oregon Health and Science University
Department
Type
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Smithey, Megan J; Venturi, Vanessa; Davenport, Miles P et al. (2018) Lifelong CMV infection improves immune defense in old mice by broadening the mobilized TCR repertoire against third-party infection. Proc Natl Acad Sci U S A 115:E6817-E6825
Maurizio, Paul L; Ferris, Martin T; Keele, Gregory R et al. (2018) Bayesian Diallel Analysis Reveals Mx1-Dependent and Mx1-Independent Effects on Response to Influenza A Virus in Mice. G3 (Bethesda) 8:427-445
Uhrlaub, Jennifer L; Smithey, Megan J; Nikolich-Žugich, Janko (2017) Cutting Edge: The Aging Immune System Reveals the Biological Impact of Direct Antigen Presentation on CD8 T Cell Responses. J Immunol 199:403-407
Pryke, Kara M; Abraham, Jinu; Sali, Tina M et al. (2017) A Novel Agonist of the TRIF Pathway Induces a Cellular State Refractory to Replication of Zika, Chikungunya, and Dengue Viruses. MBio 8:
Bottomly, Daniel; Wilmot, Beth; McWeeney, Shannon K (2015) plethy: management of whole body plethysmography data in R. BMC Bioinformatics 16:134
Gralinski, Lisa E; Ferris, Martin T; Aylor, David L et al. (2015) Genome Wide Identification of SARS-CoV Susceptibility Loci Using the Collaborative Cross. PLoS Genet 11:e1005504
Okumura, Atsushi; Rasmussen, Angela L; Halfmann, Peter et al. (2015) Suppressor of Cytokine Signaling 3 Is an Inducible Host Factor That Regulates Virus Egress during Ebola Virus Infection. J Virol 89:10399-406
LaBeaud, A Desiree; Banda, Tamara; Brichard, Julie et al. (2015) High rates of o'nyong nyong and Chikungunya virus transmission in coastal Kenya. PLoS Negl Trop Dis 9:e0003436
Mirrashidi, Kathleen M; Elwell, Cherilyn A; Verschueren, Erik et al. (2015) Global Mapping of the Inc-Human Interactome Reveals that Retromer Restricts Chlamydia Infection. Cell Host Microbe 18:109-21
Davis, Zoe H; Verschueren, Erik; Jang, Gwendolyn M et al. (2015) Global mapping of herpesvirus-host protein complexes reveals a transcription strategy for late genes. Mol Cell 57:349-60

Showing the most recent 10 out of 127 publications