Human cytomegalovirus (HCMV) is a significant public health concern in the United States. Most important are the effects of the virus on developing fetuses and immunocompromised individuals where it causes a variety of pathological conditions ranging in severity from mild to life-threatening. Since HCMV is present in a persistent or latent form in 50-90% of the world's adult population, the identification of virl gene products that contribute to the establishment and maintenance of a latent infection is an intense and important area of investigation. HCMV encodes 4 GPCR homologs including US28, which has been shown to exhibit both constitutive and agonist-dependent G-protein signaling activities in HCMV infected fibroblasts. Although US28 exhibits high level signaling, it is not essential for lytic viral replication and therefore its function in viral pathogenesis remains unclear. Interestingly, US28 mRNAs are expressed in latently infected hematopoietic progenitors and monocytes, prompting questions regarding the potential role of US28 during latent CMV infections. Our exciting preliminary data indicates that US28 promotes a successful latent infection as US28 null viruses spontaneously enter the lytic phase in hematopoietic cells where wild type virus establishes a latent infection. Therefore, we hypothesize that US28 directed signaling in HCMV infected hematopoietic progenitors leads to the establishment and/or maintenance of latency. To explore this hypothesis, we will take advantage of a newly constructed panel of US28 mutant viruses and recently established methodologies to examine US28 signaling in HCMV infected cells.
In Aim 1, we will examine the biological parameters by which US28 drives latency using viral recombinants in primary hematopoietic progenitor cells, and in Aim 2 we will examine the molecular signaling properties of US28 and determine which pathways contribute to the establishment and/or maintenance of latency. Finally, experiments like those proposed in this application are essential for our understanding of the role of CMV GPCRs in viral pathogenesis and will open novel avenues of research for the development of unique antivirals that specifically target the latent reservoir of virus infection.

Public Health Relevance

Human cytomegalovirus (HCMV) infects the majority of the population and following initial infection can reside silently in infected individuals for the duraion of their lifetime. Under a variety of circumstances, the virus can 'wake-up', or reactivate, from this silent state leading to the onset of disease. In the current study, we will examine the mechanisms by which the HCMV protein US28 functions to establish and/or maintain the silent phase with the ultimate goal of using this knowledge to destabilize the silent state and prevent disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
7R21AI119415-03
Application #
9404714
Study Section
Virology - A Study Section (VIRA)
Program Officer
Beisel, Christopher E
Project Start
2016-12-21
Project End
2017-04-30
Budget Start
2016-12-21
Budget End
2017-04-30
Support Year
3
Fiscal Year
2016
Total Cost
$136,780
Indirect Cost
$41,780
Name
Cleveland Clinic Lerner
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
135781701
City
Cleveland
State
OH
Country
United States
Zip Code
44195