Our understanding ofthe life cycles of EBV and HCMV has progressed so that we know much about their infections in tractable cell hosts including B-cells for EBV and fibroblasts for HCMV. We shall now focus on more technically demanding cell types central to the viral life cycles including epithelial cells for EBV and CD34+ cells for HCMV. EBV infects epithelial cells in vivo;it causes nasopharyngeal carcinomas, the undifferentiated form of which is uniformly EBV-positive. However, these tumors usually lose EBV upon explanting into cell culture. This loss may reflect EBV providing the tumor cells selective advantages that act only in vivo or for other unknown reasons. Conditions for infecting primary and established epithelial cells with EBV have recently been described, allowing a critical examination of EBV's life-cycle in them. We propose to characterize the dynamics of EBV visually in these tractable epithelial cells to elucidate how infections are established and maintained. In collaboration with Dr. Lambert in Project 1, we will assess the role of differentiation of epithelial cells in triggering EBV's lytic cycle. All of these experiments to visualize the life-cycle of EBV in epithelial cells will complement those to assess biochemically the latent to lytic switch in this cell type described in Project 4. We shall in parallel examine latent infections by HCMV, reactivation of which causes severe disease in the absence of a well-functioning immune system. At least one latent reservoir of HCMV is CD34+ hematopoietic progenitor cells. Primary CD34+ cells present experimental challenges such as heterogeneity and spontaneous differentiation, and thus our understanding of molecular mechanisms for the establishment, maintenance, and reactivation of HCMV latency is minimal. For example, unlike latent EBV, it is unclear if HCMV requires or even has mechanisms to replicate, maintain, and partition its genome within these cells. We have now infected established CD34+ cell lines and shown that they support latent infection of HCMV. We propose to characterize the dynamics of HCMV in these tractable CD34+ cells to elucidate how infections are established and maintained. Our collaborative studies of EBV and HCMV will use multiple approaches including live-cell imaging of visible derivatives of both viruses in these physiologically critical cell-types.

Public Health Relevance

EBV causes two kinds of carcinomas and HCMV both causes severe disease in immunocompromised patients and has been associated with glioblastoma multiforme. We have developed new techniques that allow us to study the life-cycles of these viruses in cell-types difficult to examine but critical for this pathogenesis. Our analysis of EBV and of HCMV with live-cell imaging will reveal how the viruses establish and maintain themselves in these pivotal cells and indicate how best to target them therapeutically.

Agency
National Institute of Health (NIH)
Type
Research Program Projects (P01)
Project #
5P01CA022443-37
Application #
8675201
Study Section
Special Emphasis Panel (ZCA1)
Project Start
Project End
Budget Start
Budget End
Support Year
37
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
DUNS #
City
Madison
State
WI
Country
United States
Zip Code
53715
Park, Soyeong; Park, Jung Wook; Pitot, Henry C et al. (2016) Loss of Dependence on Continued Expression of the Human Papillomavirus 16 E7 Oncogene in Cervical Cancers and Precancerous Lesions Arising in Fanconi Anemia Pathway-Deficient Mice. MBio 7:
Lee, Denis; Norby, Kathryn; Hayes, Mitchell et al. (2016) Using Organotypic Epithelial Tissue Culture to Study the Human Papillomavirus Life Cycle. Curr Protoc Microbiol 41:14B.8.1-14B.8.19
Bodelon, Clara; Vinokurova, Svetlana; Sampson, Joshua N et al. (2016) Chromosomal copy number alterations and HPV integration in cervical precancer and invasive cancer. Carcinogenesis 37:188-96
Unchwaniwala, Nuruddin; Sherer, Nathan M; Loeb, Daniel D (2016) Hepatitis B Virus Polymerase Localizes to the Mitochondria, and Its Terminal Protein Domain Contains the Mitochondrial Targeting Signal. J Virol 90:8705-19
Jones, Richard J; Iempridee, Tawin; Wang, Xiaobin et al. (2016) Lenalidomide, Thalidomide, and Pomalidomide Reactivate the Epstein-Barr Virus Lytic Cycle through Phosphoinositide 3-Kinase Signaling and Ikaros Expression. Clin Cancer Res 22:4901-4912
Zeng, Hao; Lu, Li; Chan, Ngai Ting et al. (2016) Systematic identification of Ctr9 regulome in ERα-positive breast cancer. BMC Genomics 17:902
Tan, Xiaojun; Lambert, Paul F; Rapraeger, Alan C et al. (2016) Stress-Induced EGFR Trafficking: Mechanisms, Functions, and Therapeutic Implications. Trends Cell Biol 26:352-66
Makielski, Kathleen R; Lee, Denis; Lorenz, Laurel D et al. (2016) Human papillomavirus promotes Epstein-Barr virus maintenance and lytic reactivation in immortalized oral keratinocytes. Virology 495:52-62
Chiu, Ya-Fang; Sugden, Bill (2016) Epstein-Barr Virus: The Path from Latent to Productive Infection. Annu Rev Virol 3:359-372
Chandra, Janin; Kuo, Paula T Y; Hahn, Anne M et al. (2016) Batf3 selectively determines acquisition of CD8(+) dendritic cell phenotype and function. Immunol Cell Biol :

Showing the most recent 10 out of 400 publications