Liver pathology in alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH) and forms of toxicant-associated steatohepatitis (TASH) is indistinguishable. ALD pathogenesis may be related to generation of toxic acetaldehyde (AcAld), whereas NASH and TASH are associated with generation of malondialdehyde (MDA), 4-hydroxynonenal (HNE), chloracetaldehyde (ClAcAld) and others by lipid peroxidation and/or toxicant metabolism. With the major exceptions of oxygen and short chain fatty acids, all mitochondrial metabolites cross mitochondrial outer membranes (MOM) via open voltage dependent anion channels (VDAC). The central hypothesis of this project is that ethanol and aldehydes close VDAC, decrease permeability of MOM and suppress normal mitochondrial function. Such VDAC closure allows the selective and more rapid mitochondrial oxidation of toxic aldehydes, which permeate mitochondria freely. Although VDAC closure is adaptive in promoting aldehyde detoxification, VDAC closure may be maladaptive in promoting steatosis and lipotoxicity. Accordingly, we propose to: 1) Characterize the effects of ethanol and aldehydes on ureagenesis in cultured hepatocytes, since ureagenesis is a major energy-consuming process that is dependent on exchange of metabolites across mitochondrial membranes. We will characterize the effects of ethanol, AcAld, MDA, HNE, ClAcAld and other aldehydes on ureagenic respiration and outer membrane permeability with the expectation that exogenous aldehydes and AcAld formed by ethanol metabolism will cause dose-dependent inhibition of ureagenesis and a decrease of MOM permeability to low molecular weight (? 3 kDa) solutes. Follow-on experiments identify kinase pathways mediating VDAC closure. 2) Determine the contributions of the three individual VDAC isoforms to suppression of ureagenesis in hepatocytes by ethanol and aldehydes using phosphorothioate siRNA single and double knockout/knockdowns of the VDAC isoforms. 3) Evaluate the role of aldehydes, kinases and VDAC in steatosis and lipotoxicity in vitro and in vivo. In preliminary experiments, aldehyde promoted steatosis dependent on c-Jun N-terminal kinase (JNK) after incubation of hepatocytes with Intralipid and sensitized to tumor necrosis factor-? (TNF?)-dependent apoptosis. We will evaluate mechanisms underlying this cell killing and the role of specific VDAC isoforms in promoting both steatosis and cell death. ALD and NASH are widely prevalent diseases in the U.S. for which therapy is largely ineffective. Lack of effective therapy reflects our ignorance of the underlying etiologies. In particular, the basis for the identical histopathology of ALD, NASH and TASH is unknown. Aldehyde-dependent VDAC closure provides a shared mechanism for the underlying pathophysiology of these diseases, which will likely lead to better strategies for treatment and prevention.

Public Health Relevance

Alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH) and toxiciant-associated steatohepatitis (TASH) are widely prevalent or underreported liver diseases in the U.S. that have identical histopathology, poorly understood pathophysiology and, consequently, largely ineffective therapies. This project will examine the novel hypothesis that aldehyde generation common to these diseases causes closure of voltage dependent anion channels (VDAC) in mitochondria, an event that on the one hand is adaptive in promoting mitochondrial metabolism and detoxification of freely permeant aldehydes but on the other hand is maladaptive in pre-disposing to lipotoxicity and cell death. An understanding of the shared mechanisms underlying ALD, NASH and TASH will lead to better strategies for treatment and prevention.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
1R01AA021191-01A1
Application #
8761184
Study Section
Hepatobiliary Pathophysiology Study Section (HBPP)
Program Officer
Orosz, Andras
Project Start
2014-09-15
Project End
2019-06-30
Budget Start
2014-09-15
Budget End
2015-06-30
Support Year
1
Fiscal Year
2014
Total Cost
$336,375
Indirect Cost
$111,375
Name
Medical University of South Carolina
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
183710748
City
Charleston
State
SC
Country
United States
Zip Code
29425
Maldonado, Eduardo N; Lemasters, John J (2014) ATP/ADP ratio, the missed connection between mitochondria and the Warburg effect. Mitochondrion 19 Pt A:78-84