This project seeks to determine the clinical relevance and potential role in hepatic carcinogenesis of interactions between host cell and hepatitis C virus (HCV) proteins that have been identified in previous cell culture-based experiments. The retinoblastoma susceptibility protein (pRb) and the cellular DEAD-box RNA helicase DDXS play critical roles in regulating the cell cycle. Our previous work indicates that the HCV RNA polymerase, NS5B, fc^nns a complex with pRb, targeting it for ubiquitination and proteasome-dependent degradation, activating E2F-responsive promoters, and stimulating cell proliferation. Similarly, HCV cofe protein expression alters the cellular abundance and localization of DDX3. Our hypothesis is that these and other interactions with host proteins (p53 and DDX5) promote proliferation of hepatocytes and impair DNA damage responses, tljiereby contributing to HCV carcinogenesis.
In Aim 1, we will develop and validate fluorescent quantum dot probes capable of sensitive detection of HCV proteins expressed in virus-infected cells. We will also determine whether HCV protein expression conrelates with altered tumor suppressor expression in transgenic mice.
Aim 2 will utilize these novel probes in laser scanning confocal and multi photon microscopy studies of liver tissue from patients with chronic hepatitis C, and ask whether the abundance and cellular localization of pRb, p53, DDXS, or DDXS is altered in infected hepatocytes.
In Aim 3, we will determine whether HCV infection is associated on a single-cell basis with increased expression of the proliferation markers Ki-67 and PCNA. We will also determine whether the proportion of cells displaying proliferation markers is reduced after standard-of-care peg-IFN/ribavirin therapy. These results will be correlated with oligonucleotide microarray and quantitative RT-PCR assays to determine whether liver specimens with a high proportion of HCV-infected cells display transcriptional patterns indicative of hepatocellular proliferation and/or E2F transcription factor activation.

Public Health Relevance

TO PUBLIC HEATH:,' Liver cancer is one qf the most rapidly increasing types of cancers in the United States, reflecting an increased prevalence ^nd risk of liver cancer in persons with chronic hepatitis C. This project seeks a better understanding of the interaction of HCV proteins with important regulators of cell proliferation (pRb, p53, DDXS, and DDXS) anc( the role of such interactions in liver cancer caused by HCV infection. PERFORMANCE SITE

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory Grants (P20)
Project #
5P20CA150343-03
Application #
8321047
Study Section
Special Emphasis Panel (ZCA1)
Project Start
Project End
2013-08-31
Budget Start
2011-09-01
Budget End
2012-08-31
Support Year
3
Fiscal Year
2011
Total Cost
$276,791
Indirect Cost
Name
University of Kentucky
Department
Type
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40506
Yu, T; Chen, X; Lin, T et al. (2016) KLF4 deletion alters gastric cell lineage and induces MUC2 expression. Cell Death Dis 7:e2255
Valentino, Joseph D; Li, Jing; Zaytseva, Yekaterina Y et al. (2014) Cotargeting the PI3K and RAS pathways for the treatment of neuroendocrine tumors. Clin Cancer Res 20:1212-22
Yamane, Daisuke; McGivern, David R; Wauthier, Eliane et al. (2014) Regulation of the hepatitis C virus RNA replicase by endogenous lipid peroxidation. Nat Med 20:927-35
Ye, Q; Cai, W; Zheng, Y et al. (2014) ERK and AKT signaling cooperate to translationally regulate survivin expression for metastatic progression of colorectal cancer. Oncogene 33:1828-39
Zaytseva, Yekaterina Y; Elliott, Victoria A; Rychahou, Piotr et al. (2014) Cancer cell-associated fatty acid synthase activates endothelial cells and promotes angiogenesis in colorectal cancer. Carcinogenesis 35:1341-51
Wang, Q; Zhou, Y; Rychahou, P et al. (2013) NFAT5 represses canonical Wnt signaling via inhibition of ?-catenin acetylation and participates in regulating intestinal cell differentiation. Cell Death Dis 4:e671
Valentino, Joseph D; Rychahou, Piotr G; Mustain, W Conan et al. (2013) Murine portal vein catheterization to analyze liver-directed therapies. J Surg Res 185:690-6
Li, X; Stevens, P D; Yang, H et al. (2013) The deubiquitination enzyme USP46 functions as a tumor suppressor by controlling PHLPP-dependent attenuation of Akt signaling in colon cancer. Oncogene 32:471-8
Zhang, Wen; Sviripa, Vitaliy; Chen, Xi et al. (2013) Fluorinated N,N-dialkylaminostilbenes repress colon cancer by targeting methionine S-adenosyltransferase 2A. ACS Chem Biol 8:796-803
Kim, Seungtaek; Ishida, Hisashi; Yamane, Daisuke et al. (2013) Contrasting roles of mitogen-activated protein kinases in cellular entry and replication of hepatitis C virus: MKNK1 facilitates cell entry. J Virol 87:4214-24

Showing the most recent 10 out of 30 publications