The objectives are to obtain a better understanding of the molecular effects of alcohol upon hepatic function as well as the pathogenesis of hepatic fibrosis in alcoholic liver disease. Two general areas of inquiry will be investigated: (1) What levels of gene regulation and specifically collagen gene regulation are affected in alcoholic liver disease? This will involve an analysis of mRNA steady state levels by cell free protein synthesis and RNA-DNA hybridization. Quantitative determinations of changes in several specific mRNA species including types I, III, and IV procollagen, albumin, fibronectin and -actin will be undertaken. Nuclear transcriptional run-off analysis will be performed in order to examine the basis for observed changes in steady state mRNA contents. Collagen gene promoter structure and function will be determined by constructing partial genomic libraries from cirrhotics and non-cirrhotic alcoholics, isolating and purifying the collagen promoter, and analyzing it by means of S1 nuclease sensitivity methylation, DNA sequencing, and use of a transient expression vector (pSV-0-CAT). These latter studies may explain variations in an individual's fibrotic response to alcohol. (2) What factors predispose an individual to develop alcoholic cirrhosis? This will involve a genetic analysis of the collagen gene using DNA polymorphisms and analysis of the role of sex hormones and corticosteroids in hepatic collagen synthesis using cell culture techniques, molecular hybridization studies and in vivo transient expression vector analysis. The results of such experiments will help us expand our knowledge of the molecular mechanisms responsible for hepatic fibrosis in man. This knowledge will help in developing diagnostic methods for evaluating fibrogenesis predicting susceptibility to develop cirrhosis, and provide the basis for better therapy. These studies will also facilitate my long term objective to become an independent investigator in academic hepatology with expertise in current areas of molecular biology.

Project Start
1986-08-01
Project End
1991-07-31
Budget Start
1988-08-01
Budget End
1989-07-31
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461
Weiner, F R; Shah, A; Biempica, L et al. (1992) The effects of hepatic fibrosis on Ito cell gene expression. Matrix 12:36-43
Weiner, F R; Blaner, W S; Czaja, M J et al. (1992) Ito cell expression of a nuclear retinoic acid receptor. Hepatology 15:336-42
Weiner, F R; Smith, P J; Wertheimer, S et al. (1991) Regulation of gene expression by insulin and tumor necrosis factor alpha in 3T3-L1 cells. Modulation of the transcription of genes encoding acyl-CoA synthetase and stearoyl-CoA desaturase-1. J Biol Chem 266:23525-8
Czaja, M J; Weiner, F R; Freedman, J H (1991) Amplification of the metallothionein-1 and metallothionein-2 genes in copper-resistant hepatoma cells. J Cell Physiol 147:434-8
Weiner, F R; Giambrone, M A; Czaja, M J et al. (1990) Ito-cell gene expression and collagen regulation. Hepatology 11:111-7
Czaja, M J; Flanders, K C; Biempica, L et al. (1989) Expression of tumor necrosis factor-alpha and transforming growth factor-beta 1 in acute liver injury. Growth Factors 1:219-26
Weiner, F R; Shah, A; Smith, P J et al. (1989) Regulation of collagen gene expression in 3T3-L1 cells. Effects of adipocyte differentiation and tumor necrosis factor alpha. Biochemistry 28:4094-9