studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 265A
decompensation, reduced risk for hepatocellular carcinoma,
and reduced liver-related mortality. We propose that cure of
CHC (defined as SVR of at least 12 weeks) can also lead to
regression of advanced fibrosis and/or cirrhosis and that Fibroscan
elastography may be used to confirm reversal of fibrosis.
METHODS: We conducted a retrospective chart review to
identify patients treated for CHC with concomitant advanced
fibrosis or cirrhosis based on clinical features, Fibroscan scores
and/or biopsy. Advanced fibrosis was defined as FibroScan
score 11-13.9Kpa and cirrhosis was defined as Fibroscan
score > 14KPa. This was followed by prospective and retrospective
Fibroscan and clinical data collection at 6 month intervals,
up to 14 years. RESULTS: 201 subjects were enrolled, 58
are currently eligible for analysis with at least 2 measurements
of fibrosis. Mean age was 60.7 years, 70.7% were male, and
most were Non-Hispanic (81%) white (91.4%). At baseline
25.9% had hypertension, 12.1 % had diabetes, 12.1% had
hyperlipidemia and the average BMI was 27.7 (SD 5). Of
the 34 subjects who had cirrhosis at baseline, 18 (52.9%)
demonstrated improvement by Fibroscan, with a median time
to improvement of 2.8 years (IQR 1.0-3.5). Improvement was
defined as a change in stage of fibrosis rather than a change in
Fibroscan score. Of the 17 subjects who had advanced fibrosis,
16 (66.7%) demonstrated improvement, with a median time of
2.0 years (IQR 1.2-2.5). Gender, BMI, age, individual medical
problems and HCV genotype were not found to be associated
with improvement or worsening of baseline liver disease. ALT,
albumin and INR changes did not correlate with fibrosis regression.
However, platelet count change (increased counts in subjects
who demonstrated improved liver disease) approached
statistical significance with a p=0.06 in subjects with cirrhosis.
CONCLUSION: The majority of our subjects, 34/58 (58.6%),
with advanced fibrosis or cirrhosis demonstrated improvement
by Fibroscan scores after a median follow-up (after treatment
of CHC) of 2.5 years (range: 0.5-14 years, IQR: 1- 3.5 years).
We have yet been not able to establish factors that may predict
improvement of cirrhosis or advanced fibrosis. Most enrolled
patients are still awaiting follow-up Fibroscan test results anticipated
to be completed in 2015.
Disclosures:
Catherine T. Frenette - Speaking and Teaching: Bayer, Salix, Gilead; Stock
Shareholder: Gilead
Paul J. Pockros - Advisory Committees or Review Panels: Janssen, Merck, BMS,
Gilead, AbbVie; Consulting: Lumena, Beckman Coulter; Grant/Research Support:
Intercept, Janssen, BMS, Gilead, Lumena, Beckman Coulter, AbbVie, RMS,
Merck; Speaking and Teaching: AbbVie, Janssen, Gilead
The following authors have nothing to disclose: Ana Maria Crissien, William B.
Minteer, Jason J. Pan
109
Circulating extracellular vesicles and their microRNA
cargos are potential novel biomarkers with a functional
role of miR-122 in monocyte activation in alcoholic hepatitis
Banishree Saha, Fatemah Momen-Heravi, Shashi Bala, Karen
Kodys, Gyongyi Szabo; Medicine, UMASS Med School, Worcester,
MA
Purpose: Alcohol and its metabolites induce hepatocyte damage
and recruitment of inflammatory monocytes/macrophages
and neutrophils leading to alcoholic hepatitis. Currently there
is no reliable biomarker of alcoholic hepatitis. Extracellular
vesicles (EVs) found in the circulation carry mRNA, microRNA
(miRs) and proteins. EVs can serve as biomarkers and mediate
intercellular communication. miR-122 is abundantly expressed
in hepatocytes, not in immune cells and increased levels of circulating
miR-122 were found in liver injury. We hypothesized
that EV-associated miRs can serve as biomarkers and modulate
intercellular signaling between hepatocytes and immune
cells in alcoholic hepatitis. Methods: EVs were isolated from
chronic alcohol-fed (5 weeks of Lieber DeCarli diet) or pair-fed
mice sera and from serum of patients with alcoholic hepatitis.
EVs were characterized by transmission electron microscopy,
western blot, nanoparticle tracking analysis system and
miRNA analysis. Results: The total number of circulating EVs
was significantly increased in alcohol-fed mice as compared
to control mice. Exosomes (40-150nm) represented most of
the EVs (~80%). MicroRNA array of circulating EVs revealed
a significant increase of 7 inflammatory miRs including: miR-
192, 122, 30a, 744, 1246, 30b and miR-130a in alcohol-fed
mice compared to controls. The ROC analyses indicated excellent
diagnostic value of miR-192, 122, and 30a to identify
alcohol-induced liver injury. In patients with acute alcoholic
hepatitis, we found a significant increase in the number of
circulating EVs compared to normal controls and miR-192 and
miR-30a were significantly increased in the EVs from alcoholic
hepatitis patients. Serum miR-122 was increased after alcohol
binge drinking. In the liver, miR-122 is abundantly expressed
in hepatocytes and monocytes/macrophages have low levels.
In vitro experiments revealed that exosomes derived from
ethanol-treated human hepatocytes were taken up by monocytes
and transferred mature miR-122 into monocytes. This horizontally
transferred miR-122 inhibited the hemeoxygenase-1
expression, a target of miR-122 and sensitized monocytes to
LPS stimulation to increase production of pro-inflammatory cytokines,
TNF-α and IL-1β; all of these effects were inhibited by
exosome-mediated delivery of a miR-122 inhibitor in monocytes.
Conclusion: Elevated levels of EVs and their miR signature
could serve as biomarkers of alcoholic hepatitis. This study
reveals a novel EV-mediated mechanism of alcohol-induced
communication between hepatocytes and monocytes by transferring
hepatocyte-derived miR-122 that reprograms monocytes
promoting inflammation in alcoholic hepatitis.
Disclosures:
The following authors have nothing to disclose: Banishree Saha, Fatemah
Momen-Heravi, Shashi Bala, Karen Kodys, Gyongyi Szabo
110
Fat-specific Protein 27/CIDEC Promotes Alcoholic Steatohepatitis
in Mice and Humans
Ming-Jiang Xu 1 , Yan Cai 1 , Hua Wang 1 , José T. Altamirano 2 ,
Gemma Odena 3 , Frank J. Gonzalez 4 , Ramon Bataller 3 , Bin Gao 1 ;
1 NIAAA, NIH, Rockville, MD; 2 Vall d’Hebron Institut de Recerca,
Barcelona, Spain; 3 University of North Carolina, Chapel Hill, NC;
4 National Institutes of Health, Bethesda, MD
Objectives: Alcoholic steatohepatitis (ASH) is the progressive
form of alcoholic liver disease that leads to cirrhosis and
hepatocellular carcinoma. There is an urgent need to identify
molecular drivers in order to develop targeted therapies.
Here the functions of mouse fat-specific protein 27 (Fsp27)/
human cell death activator CIDEC (the human homologue of
Fsp27) were investigated. Methods and results: We developed
a mouse model with chronic (8 weeks)-plus-binge ethanol feeding,
which mimics the drinking patterns of alcoholic hepatitis
(AH) patients, produced severe ASH and mild fibrosis. Microarray
analyses revealed that mouse Fsp27/human CIDEC gene
was increased in this animal model and human ASH samples.
Fsp27 is expressed at high levels in adipose tissues but at
very low levels in normal liver. Chronic-plus-binge ethanol feeding
markedly upregulated hepatic Fsp27 mRNA and protein
expression. Silencing the Fsp27 gene by shRNA or genetic
disruption ameliorated chronic-plus-binge ethanol-induced