The objective of this proposal is to determine the involvement of the lipid phosphatase PTEN in the formation of steatosis and liver damage during alcoholic liver disease (ALD). Although there are publications concerning the role of PTEN in ALD, these reports are using a static time point in a fluid model and do not address the possibility of both PTEN inactivation via aldehyde modification and changes in PTEN regulation/expression over a period of alcohol exposure. Our working hypothesis states the reactive aldehyde 4-HNE, produced in the liver of chronic ethanol treated mice covalently adducts PTEN thereby reducing enzymatic activity leading to disregulation in PTEN downstream signaling. 4-HNE has already been implicated in the inactivation of proteins during chronic ethanol exposure in the liver. It is also hypothesized that increased PTEN expression leads to changes in downstream pathways ultimately leading to increased steatosis seen in ALD. In order to fulfill the objectives of this research proposal, experiments will be performed using mass spectrometry to identify the sites of 4-HNE modification on PTEN, the ability of these adducts to inhibit enzymatic activity, membrane association and Trx1 association. In addition, using western blotting, immunohistochemistry and mRNA expression, the effects of variable PTEN expression on downstream signaling pathways will be examined following 9-weeks of chronic ethanol exposure.

Public Health Relevance

In the United States today, alcohol induced liver disease is a major cause of morbidity and mortality. Current statistics indicate that among chronic alcohol users, 15% will eventually be diagnosed with alcoholic liver disease. These symptoms include steatosis, alcoholic hepatitis and cirrhosis. The 5 and 10 year survival rates for alcohol induced cirrhosis are 23% and 7% respectively. Completion of the proposed experiments will provide greater insight into the mechanisms of chronic alcohol toxicity especially with respect to steatosis and damage caused by increased oxidative stress and altered PTEN signaling.

National Institute of Health (NIH)
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Postdoctoral Individual National Research Service Award (F32)
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Health Services Research Review Subcommittee (AA)
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Radaeva, Svetlana
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University of Colorado Denver
Schools of Pharmacy
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Petersen, Dennis R; Saba, Laura M; Sayin, Volkan I et al. (2018) Elevated Nrf-2 responses are insufficient to mitigate protein carbonylation in hepatospecific PTEN deletion mice. PLoS One 13:e0198139
Shearn, Colin T; Saba, Laura M; Roede, James R et al. (2017) Differential carbonylation of proteins in end-stage human fatty and nonfatty NASH. Free Radic Biol Med 113:280-290
Shearn, Colin T; Orlicky, David J; McCullough, Rebecca L et al. (2016) Liver-Specific Deletion of Phosphatase and Tensin Homolog Deleted on Chromosome 10 Significantly Ameliorates Chronic EtOH-Induced Increases in Hepatocellular Damage. PLoS One 11:e0154152
Shearn, C T; Orlicky, D J; Saba, L M et al. (2015) Increased hepatocellular protein carbonylation in human end-stage alcoholic cirrhosis. Free Radic Biol Med 89:1144-53
Shearn, Colin T; Petersen, Dennis R (2015) Understanding the tumor suppressor PTEN in chronic alcoholism and hepatocellular carcinoma. Adv Exp Med Biol 815:173-84
Shearn, Colin T; Mercer, Kelly E; Orlicky, David J et al. (2014) Short term feeding of a high fat diet exerts an additive effect on hepatocellular damage and steatosis in liver-specific PTEN knockout mice. PLoS One 9:e96553
Shearn, Colin T; Backos, Donald S; Orlicky, David J et al. (2014) Identification of 5' AMP-activated kinase as a target of reactive aldehydes during chronic ingestion of high concentrations of ethanol. J Biol Chem 289:15449-62
Shearn, C T; Smathers, R L; Backos, D S et al. (2013) Increased carbonylation of the lipid phosphatase PTEN contributes to Akt2 activation in a murine model of early alcohol-induced steatosis. Free Radic Biol Med 65:680-692
Shearn, Colin T; Smathers, Rebecca L; Jiang, Hua et al. (2013) Increased dietary fat contributes to dysregulation of the LKB1/AMPK pathway and increased damage in a mouse model of early-stage ethanol-mediated steatosis. J Nutr Biochem 24:1436-45
Shearn, Colin T; Reigan, Philip; Petersen, Dennis R (2012) Inhibition of hydrogen peroxide signaling by 4-hydroxynonenal due to differential regulation of Akt1 and Akt2 contributes to decreases in cell survival and proliferation in hepatocellular carcinoma cells. Free Radic Biol Med 53:1-11

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