In characterizing the cell-cycle regulatory role of the cyclin-dependent kinase inhibitor, p27kip1, it was discovered that p27 knockout mice exhibit pronounced multi-organomegaly with no increased spontaneous carcinogenesis in the liver. This suggested that p27 manipulation could improve the success of hepatocyte transplantation. It was then shown that donor hepatocytes from p27 knockout mice more actively proliferate and rescue liver failure in the FAH mouse model. These results were limited by the fact that p27 was suppressed throughout the donor animal from early embryogenesis and the risk of carcinogenesis due to longterm p27 absence was not determined. In this proposal we develop two new techniques for targeting p27 in the mouse liver. Mx 1-Cre transgenic mice will allow us to knockout p27 in the liver alone at controlled time points of hepatogenesis. Ectopic expression of the F-box protein Skp2 from Alumin promoter will allow us to lower p27 protein abundance in the hepatocytes. We also propose to inactivate p27 through lentivirus mediated delivery of Skp2 and p27 specific shRNA, which can be directly applicable to human hepatocytes in the future. With these new approaches of p27 inactivation, we will determine the role of p27 in hepatocyte transplantation and hepatocellular carcinogenesis in more clinically relevant settings. The results will give us a clear assessment of the potential benefits of p27 suppression in the treatment of human liver failure.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08DK065880-04
Application #
7448550
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Program Officer
Podskalny, Judith M,
Project Start
2005-07-01
Project End
2010-06-30
Budget Start
2008-07-01
Budget End
2009-06-30
Support Year
4
Fiscal Year
2008
Total Cost
$126,630
Indirect Cost
Name
University of Pittsburgh
Department
Pathology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Limaye, Pallavi B; Alarcon, Gabriela; Walls, Andrew L et al. (2008) Expression of specific hepatocyte and cholangiocyte transcription factors in human liver disease and embryonic development. Lab Invest 88:865-72
Song, Joon Seok; Murase, Noriko; Demetris, Anthony J et al. (2007) Protection from acute cellular injury using Sleeping Beauty mediated telomerase gene transfer. Biochem Biophys Res Commun 363:253-6