Kaposi's sarcoma-associated herpesvirus (KSHV) encodes 12 pre-miRNAs that are highly expressed in KS lesions and KSHV- associated lymphoproliferative malignancies. Ectopic expression and miRNA inhibition approaches, combined with gene expression profiling, have identified target mRNAs that are involved in the regulation of critical biological processes such as angiogenesis, apoptosis, endothelial cell differentiation, and immune surveillance. In addition, several miRNAs may regulate latency by targeting viral mRNAs. Preliminary analyses of the mRNAs targeted by KSHV miRNAs in PEL cells using HITS-CLIP technology to enrich RISC-associated RNAs have identified hundreds of new mRNA targets and confirmed previously identified targets. Moreover, our analysis showed that a high percentage of RISC complexes within PEL cells contain KSHV- encoded miRNAs. Based on these data, we hypothesize that the KSHV miRNAs cause significant and tissue-specific changes in gene expression that contribute to pathogenesis and tumorigenesis. To directly address this hypothesis we propose to 1) Comprehensively identify KSHV miRNA targets by HITS-CLIP in KSHV-infected PEL and endothelial cells;2) Characterize in vitro phenotypes and their associated regulatory pathways with respect to viral replication, the establishment and maintenance of latency, cell cycle control and resistance to apoptosis by using KSHV mutants lacking single or multiple miRNAs. In summary, combining ribonomics approaches for miRNA target identification with viral genetics, and the ability to determine viral phenotypes in cultured cells, will provide a detailed analysis of KSHV-encoded miRNA functions and may lead to the development of antiviral/antitumor strategies.

Public Health Relevance

Human Herpesviruses cause many diseases including cancer and recently have been found to express miRNAs. Understanding how miRNA regulation of gene expression contributes to viral biology and pathogenesis can be important for the development of antiviral strategies.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA119917-07
Application #
8620613
Study Section
Special Emphasis Panel (ZRG1-AARR-K (03))
Program Officer
Read-Connole, Elizabeth Lee
Project Start
2005-12-01
Project End
2018-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
7
Fiscal Year
2014
Total Cost
$270,967
Indirect Cost
$89,819
Name
University of Florida
Department
Genetics
Type
Schools of Medicine
DUNS #
969663814
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Verma, Dinesh; Li, Da-Jiang; Krueger, Brian et al. (2015) Identification of the physiological gene targets of the essential lytic replicative Kaposi's sarcoma-associated herpesvirus ORF57 protein. J Virol 89:1688-702
Feldman, Emily R; Kara, Mehmet; Coleman, Carrie B et al. (2014) Virus-encoded microRNAs facilitate gammaherpesvirus latency and pathogenesis in vivo. MBio 5:e00981-14
Haecker, Irina; Renne, Rolf (2014) HITS-CLIP and PAR-CLIP advance viral miRNA targetome analysis. Crit Rev Eukaryot Gene Expr 24:101-16
Yang, Yajie; Boss, Isaac W; McIntyre, Lauren M et al. (2014) A systems biology approach identified different regulatory networks targeted by KSHV miR-K12-11 in B cells and endothelial cells. BMC Genomics 15:668
Reese, T A; Wakeman, B S; Choi, H S et al. (2014) Helminth infection reactivates latent ?-herpesvirus via cytokine competition at a viral promoter. Science 345:573-7
Plaisance-Bonstaff, Karlie; Choi, Hong Seok; Beals, Tyler et al. (2014) KSHV miRNAs decrease expression of lytic genes in latently infected PEL and endothelial cells by targeting host transcription factors. Viruses 6:4005-23
Zhu, Ying; Haecker, Irina; Yang, Yajie et al. (2013) ?-Herpesvirus-encoded miRNAs and their roles in viral biology and pathogenesis. Curr Opin Virol 3:266-75
Han, Soo-Jin; Marshall, Vickie; Barsov, Eugene et al. (2013) Kaposi's sarcoma-associated herpesvirus microRNA single-nucleotide polymorphisms identified in clinical samples can affect microRNA processing, level of expression, and silencing activity. J Virol 87:12237-48
Boss, Isaac W; Renne, Rolf (2011) Viral miRNAs and immune evasion. Biochim Biophys Acta 1809:708-14
Boss, Isaac W; Nadeau, Peter E; Abbott, Jeffrey R et al. (2011) A Kaposi's sarcoma-associated herpesvirus-encoded ortholog of microRNA miR-155 induces human splenic B-cell expansion in NOD/LtSz-scid IL2Rýýnull mice. J Virol 85:9877-86

Showing the most recent 10 out of 14 publications