Severe alcoholic liver disease (ALD) has a high morbidity and mortality. Recent studies demonstrated that probiotics reversed alcohol-induced hepatic steatosis and inflammation, and improved liver enzymes in animal models and in patients. Our laboratory showed that administration of a probiotic strain, Lactobacillus rhamnosus Gorbach-Goldin (LGG), significantly improved liver enzymes and histology in alcohol treated mice. These mice also had reduced intestinal permeability and plasma endotoxin levels. Gut-derived endotoxins normally penetrate the gut epithelium only in trace amounts due to tightly regulated intestinal barrier function. However, endotoxin leakiness is increased under pathological conditions, including alcohol abuse. We recently reported that chronic alcohol intake in mice caused a marked reduction in the intestinal expression of the master transcription factor, hypoxia inducible factor (HIF), and two HIF related proteins, cathelin-related antimicrobial peptide (CRAMP) and intestinal trefoil factor (ITF), in the intestine. ITF plays a critical role in the formation and stabilization of the intestinal mucus barrier, enhancement of rapid intestinal repair (restitution), and modulation of mucosal differentiation processes and immune responses, while CRAMP is important in blocking intestinal bacterial overgrowth/dysbiosis. In addition, our preliminary data demonstrate a significant decrease of short chain fatty acids (SCFAs), in particular, butyric acid, in the intestinal lumen following alcohol exposure in rodents. Our hypothesis is that alcohol consumption causes decreased intestinal CRAMP and ITF expression and intestinal lumen butyric acid concentration, resulting in bacterial overgrowth/dysbiosis and disrupted gut-barrier function; LGG supplementation attenuates these deleterious effects and protects the liver from alcohol-induced injury. To test this hypothesis, we will have 3 specific aims.
Specific aim 1 will determine the role of CRAMP and the beneficial effects of LGG on CRAMP in alcohol-mediated changes in gut microbiota homeostasis. The experiments will be performed using a metagenomic approach in CRAMP knockout mice.
Specific aim 2 will determine the role of LGG in promoting ITF expression and will evaluate whether ITF positively modulates intestinal tight junctions as a mechanism for improved intestinal barrier integrity. We will use goblet and intestinal epithelial cell culture, and recombinant ITF to delineate the effect of ITF.
Specific aim 3 will determine the role and mechanisms of butyric acid in the beneficial effects of LGG on intestinal tight junctions using a butyric transporter knockout mouse model. This study will have a major impact on the development of probiotic-based new therapeutic strategy for the prevention and treatment of ALD.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Exploratory Grants (P20)
Project #
1P20GM113226-01
Application #
8813883
Study Section
Special Emphasis Panel (ZGM1-TWD-A (C1))
Project Start
2016-06-10
Project End
2021-03-31
Budget Start
2014-12-01
Budget End
2015-11-30
Support Year
1
Fiscal Year
2016
Total Cost
$184,680
Indirect Cost
$64,680
Name
University of Louisville
Department
Type
DUNS #
057588857
City
Louisville
State
KY
Country
United States
Zip Code
40208
Ghosh Dastidar, Shubha; Warner, Jeffrey B; Warner, Dennis R et al. (2018) Rodent Models of Alcoholic Liver Disease: Role of Binge Ethanol Administration. Biomolecules 8:
Gosney, Julie A; Wilkey, Daniel W; Merchant, Michael L et al. (2018) Proteomics reveals novel protein associations with early endosomes in an epidermal growth factor-dependent manner. J Biol Chem 293:5895-5908
Atay, Safinur; Wilkey, Daniel W; Milhem, Mohammed et al. (2018) Insights into the Proteome of Gastrointestinal Stromal Tumors-Derived Exosomes Reveals New Potential Diagnostic Biomarkers. Mol Cell Proteomics 17:495-515
Schuster, Susanne; Johnson, Casey D; Hennebelle, Marie et al. (2018) Oxidized linoleic acid metabolites induce liver mitochondrial dysfunction, apoptosis, and NLRP3 activation in mice. J Lipid Res 59:1597-1609
Lang, Anna L; Chen, Liya; Poff, Gavin D et al. (2018) Vinyl chloride dysregulates metabolic homeostasis and enhances diet-induced liver injury in mice. Hepatol Commun 2:270-284
Pandit, Harshul; Li, Yan; Li, Xuanyi et al. (2018) Enrichment of cancer stem cells via ?-catenin contributing to the tumorigenesis of hepatocellular carcinoma. BMC Cancer 18:783
Das, Samarendra; Rai, Anil; Mishra, D C et al. (2018) Statistical Approach for Gene Set Analysis with Trait Specific Quantitative Trait Loci. Sci Rep 8:2391
He, Liqing; Prodhan, Md Aminul Islam; Yuan, Fang et al. (2018) Simultaneous quantification of straight-chain and branched-chain short chain fatty acids by gas chromatography mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 1092:359-367
Vatsalya, Vatsalya; Kong, Maiying; Cave, Matthew C et al. (2018) Association of serum zinc with markers of liver injury in very heavy drinking alcohol-dependent patients. J Nutr Biochem 59:49-55
Prodhan, Md Aminul Islam; Yin, Xinmin; Kim, Seongho et al. (2018) Surface fitting for calculating the second dimension retention index in comprehensive two-dimensional gas chromatography mass spectrometry. J Chromatogr A 1539:62-70

Showing the most recent 10 out of 68 publications