The long term goal of this research project is to elucidate the molecular and cellular developments of hepatocellular carcinoma (HCC) in rats by applying in vitro techniques of gene modification and clonal selection (cell selection/ clonal amplification) of cells with in vivo clonal analysis of lesion development after transplantation of modified cells into liver of host rats. In vivo studies are based on the classic experimental model of carcinogenesis in rats, and the cells used are WB-F344 rat liver epithelial cells which have stem-like properties and differentiate into hepatocytes when transplanted into livers of syngeneic host rats. Liver epithelial cells that defined express aberrations in the structure and/or expression of genes normally involved in the regulation of cell proliferation, death, and differentiation have been clonally isolated from tumorigenic populations transformed spontaneously or by exposure to a carcinogenic chemical in vitro, in work on this project. Additional clonal lines will be genetically engineered to contain mutations or deletions and/or constitutively express selected genes or gene combinations involved in regulating cell proliferation, death, and differentiation. Cells bearing known gene aberrations will be transplanted into syngeneic host rats in order to assess their ability to grow progressively to form specific preneoplastic lesions and/or HCC in the regulating environment of the liver. In work done on this project we have already shown that the microenvironment regulates liver epithelial cells unlike the subcutaneous (SC) and intraperitoneal (IP) microenvironments; some lines of aneuploid liver epithelial transformants that are highly tumorigenic in SC or IP sites differentiate into hepatocyte-like cells in the liver, in coordination with slowed proliferation, increased death, and delayed or suppressed tumorigenicity. Dr Grisham believes that by coupling clonal analysis of preneoplastic and neoplastic cellular lesions that develop in the liver from transplanted cells that contain specific aberrations in gene function, he can identify the specific genes whose abnormal functions are responsible for clonal emergence of specific preneoplastic lesions and HCC, and thereby delineate the molecular pathogenesis of HCC.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA029323-21
Application #
6375632
Study Section
Pathology B Study Section (PTHB)
Program Officer
Yang, Shen K
Project Start
1981-02-01
Project End
2003-04-30
Budget Start
2001-05-01
Budget End
2002-04-30
Support Year
21
Fiscal Year
2001
Total Cost
$314,751
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Pathology
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Gordon, Gavin J; Butz, Genelle M; Grisham, Joe W et al. (2002) Isolation, short-term culture, and transplantation of small hepatocyte-like progenitor cells from retrorsine-exposed rats. Transplantation 73:1236-43
Grisham, Joe W; Coleman, William B (2002) Molecular regulation of hepatocyte generation in adult animals. Am J Pathol 161:1107-10
Malouf, N N; Coleman, W B; Grisham, J W et al. (2001) Adult-derived stem cells from the liver become myocytes in the heart in vivo. Am J Pathol 158:1929-35
Kaufmann, W K; Behe, C I; Golubovskaya, V M et al. (2001) Aberrant cell cycle checkpoint function in transformed hepatocytes and WB-F344 hepatic epithelial stem-like cells. Carcinogenesis 22:1257-69
Gordon, G J; Coleman, W B; Hixson, D C et al. (2000) Liver regeneration in rats with retrorsine-induced hepatocellular injury proceeds through a novel cellular response. Am J Pathol 156:607-19
Gordon, G J; Coleman, W B; Grisham, J W (2000) Bax-mediated apoptosis in the livers of rats after partial hepatectomy in the retrorsine model of hepatocellular injury. Hepatology 32:312-20
Gordon, G J; Coleman, W B; Grisham, J W (2000) Induction of cytochrome P450 enzymes in the livers of rats treated with the pyrrolizidine alkaloid retrorsine. Exp Mol Pathol 69:17-26
Gordon, G J; Coleman, W B; Grisham, J W (2000) Temporal analysis of hepatocyte differentiation by small hepatocyte-like progenitor cells during liver regeneration in retrorsine-exposed rats. Am J Pathol 157:771-86
Coleman, W B; Ricketts, S L; Borchert, K M et al. (1999) Induction of rat WT1 gene expression correlates with human chromosome 11p11.2-p12-mediated suppression of tumorigenicity in rat liver epithelial tumor cell lines. Int J Oncol 14:957-63
McCullough, K D; Coleman, W B; Ricketts, S L et al. (1998) Plasticity of the neoplastic phenotype in vivo is regulated by epigenetic factors. Proc Natl Acad Sci U S A 95:15333-8

Showing the most recent 10 out of 44 publications