studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 873A
decreased levels of genes associated with wound healing,
MMP regulation and angiogenesis. Quantitative PCR demonstrated
a significant decrease in alpha-SMA and TIMP3 and a
significant increase in let-7a in Lin28a mutant ethanol treated
mice compared to Lin28a mutant controls. Furthermore, H&E
staining of liver sections showed increased vascularization
and steatosis in all animals treated with ethanol compared
to untreated animals. Significantly reduced Sirius red staining
was observed in ethanol treated Lin28a mutant mice relative
to WT ethanol controls, and in Lin28a mutant mice compared
to wild type animals. Conclusions: Lin28a/let-7 axis regulates
innate inflammation and fibrotic prone processes and diminishes
alcoholic liver injury. Modulation of Lin28a/let-7 system
could be the potential therapeutic approach for alcoholic liver
disease.
Disclosures:
The following authors have nothing to disclose: Kelly McDaniel, Tami Annable,
Yuyan Han, Tianhao Zhou, Julie Venter, Ying Wan, Jessica S. Garner, Nan Wu,
Shannon S. Glaser, Heather L. Francis, Gianfranco Alpini, Fanyin Meng
1350
Elevated leukocyte count combined with radiologic evidence
of a nodular liver surface in uninfected patients
strongly predicts histologic alcoholic hepatitis, obviating
the need for liver biopsy
Nitzan Roth 1 , Behnam Saberi 2 , Jared Macklin 3 , John Donovan 1 ,
Andrew Stolz 1 , Gary C. Kanel 4 , Neil Kaplowitz 1 ; 1 Division of GI/
Liver Diseases, University of Southern California, Los Angeles, CA;
2 Division of Gastroenterology and Hepatology, The Johns Hopkins
University, Baltimore, MD; 3 Department of Medicine, University of
Southern California, Los Angeles, CA; 4 Department of Pathology,
University of Southern California, Los Angeles, CA
Background: The clinical presentation of alcoholic hepatitis
(AH) can be mimicked by other alcoholic liver diseases. Our
aim was to identify the clinical features that predict having histologic
evidence of AH in order to define a profile that would
obviate the need for liver biopsy. Methods: An expert hepatopathologist
blinded to clinical data retrospectively reviewed
the liver biopsies of 95 active alcoholic patients hospitalized
for presumed severe AH with a discriminant function of >=32
and a bilirubin level of >=5 mg/dL. Patients were defined as
having definite histologic evidence of AH if lobular inflammation
and hepatocyte damage as indicated by moderate to
severe ballooning or easily seen Mallory-Denk bodies were
found. Patients were defined as having no AH if there was
no ballooning or Mallory-Denk bodies and absent or minimal
lobular inflammation. Patients were defined as having possible
AH if they did not meet the histologic criteria for either definite
AH or no AH. Results: Fifty patients (53%) had definite AH and
33 patients (35%) had possible AH. Of the 12 patients (12%)
without AH, five had cirrhosis; the other seven with non-cirrhotic
liver disease had alcoholic foamy degeneration or alcoholic
fatty liver with cholestasis. Thirty-day survival was 94%
for definite AH, 91% for possible AH, and 100% for those with
no AH. Liver surface nodularity on imaging was 98% sensitive
and 38% specific for histologic cirrhosis. In an analysis limited
to uninfected patients with definite AH or no AH, greater leukocyte
count at admission and liver surface nodularity were
independent predictors of definite AH on biopsy (P=14x10 9 /L had
a sensitivity of 43% and a specificity of 100% for diagnosing
definite AH. If a nodular liver surface was present, a leukocyte
count of >=10x10 9 /L had a sensitivity of 80% and a specificity
of 100% for diagnosing definite AH. Using these cut offs,
44% of our cohort met clinical criteria for AH, including 12
of the 33 patients (36%) initially classified as possible AH on
biopsy. Conclusions: The combination of an elevated leukocyte
count and a nodular liver surface in the absence of infection
retrospectively identified patients with a high likelihood of having
histologic AH and suggests that a liver biopsy may not be
necessary in this subset. For patients with suspected severe AH
who do not fulfill these criteria, liver biopsy remains important,
as 23% of these patients have no histologic AH.
Disclosures:
The following authors have nothing to disclose: Nitzan Roth, Behnam Saberi,
Jared Macklin, John Donovan, Andrew Stolz, Gary C. Kanel, Neil Kaplowitz
1351
Molecular chaperone Hsp90 regulates cell-specific
induction of epigenetic genes important in macrophage
activation and lipid metabolism during alcoholic liver
injury
Pranoti Mandrekar, Aditya Ambade, Arlene Lim; Medicine, University
of Massachusetts Medical School, Worcester, MA
Background and Aims: Acetylation and methylation of chromatin-modifying
enzymes and epigenetic genes by alcohol
influence transcriptional activity of genes and impact alcoholic
liver disease (ALD). Cellular stress induced molecular chaperones
particularly hsp90 are being recognized as mediators of
environmental impacts on epigenetic states and transgenerational
inheritance. We hypothesized that chronic alcohol exposure
modulates expression of chromatin modifying enzymes
chaperoned by hsp90 during liver injury. Methods: To test
our hypothesis, C57BL/6 mice were subjected to the NIAAA-
Gao chronic-binge alcohol feeding model using Lieber-deCarli
diet with 5% v/v ethanol. A single injection of hsp90 inhibitor,
17-DMAG [17-Dimethylamino-ethylamino-17-demethoxygeldanamycin]
was administered (30-50 mg/kg BW) i.p.
before the binge. Serum ALT, liver cytokine expression and
steatosis were assessed. Hsp90α was estimated in liver nuclear
fractions. Chromatin modifying enzyme expression was analyzed
in whole livers, isolated hepatocytes and Kupffer cells
using PCR arrays. Results: Gene ontology analysis reveals
alterations in histone acetyltransferase, histone methyltransferase,
SET domain proteins and histone deacetylases in the
alcoholic liver. Five genes upregulated include ATF2 (histone
acetyltransferase), PRMT6 and SETD7 (histone methyltransferases),
RPS6KA3 (kinase) and HDAC3 (histone deacetylase)
whereas HDAC9 (histone deacetylase) was downregulated
during ALD. Analysis of expression in isolated hepatocytes and
Kupffer cells exhibits cell type specific regulation of expression.
ATF2 was exclusively upregulated (2 fold) in alcohol exposed
Kupffer cells while PRMT6 (3.2 fold) increased in hepatocytes.
HDAC9, on the other hand, was down regulated (7 fold) in
alcohol exposed hepatocytes. HDAC3, SETD7 and RPS6KA3
were increased in KCs and hepatocytes. Inhibition of hsp90
using 17-DMAG after chronic alcohol exposure significantly
alleviated liver injury measured by reduced serum ALT and liver
triglycerides. 17-DMAG altered expression of ATF2, PRMT6,
HDAC3 and HDAC9 in the liver. Phosphorylation of ATF2 in
LPS stimulated macrophages activates transcription of pro-inflammatory
cytokines. 17-DMAG treatment after alcohol feeding
prevented an increase in ATF2 expression (P