studies

624A AASLD ABSTRACTS HEPATOLOGY, October, 2015
835
Knockdown of the melatonin receptor, MT1, reduces
biliary hyperplasia and liver fibrosis in cholestatic bile
duct ligated (BDL) mice
Nan Wu 2 , Fanyin Meng 1 , Ying Wan 3 , Julie Venter 2 , Holly A. Standeford
4 , Heather L. Francis 1 , Lindsey Kennedy 4 , Yuyan Han 1,2 ,
Kelly McDaniel 3 , Antonio Franchitto 5 , Paolo Onori 5 , Sharon
DeMorrow 1 , Tami Annable 3 , Eugenio Gaudio 5 , Shannon S. Glaser
1 , Gianfranco Alpini 1 ; 1 Research, S&W DDRC and Medicine,
Central Texas Veterans Health Care System, Scott & White and
Texas A & M Health Science Center College of Medicine and
Scott White, Temple, TX; 2 Medicine, Texas A&M University HSC,
Temple, TX; 3 Operational Funds, BaylorScott&White, Temple, TX;
4 Research, Central Texas Veterans Health Care System, Temple,
TX; 5 Department of Anatomical, Histological, Forensic Medicine
and Orthopedics Sciences, University La Sapienza, Rome, Italy
Cholestatic liver diseases are characterized by increased biliary
proliferation and intrahepatic bile duct mass (IBDM), liver
damage and fibrosis. Melatonin, which is synthesized in the
pineal gland as well as cholangiocytes, exerts its effects by
interaction with MT1 and MT2 receptors. Knockdown of MT2
increases biliary proliferation and liver fibrosis in cholestatic
mice. Also, melatonin inhibits biliary hyperplasia by interacting
with MT1 receptors. However, since these studies were
performed only in vitro, we evaluated the role of MT1 in regulating
biliary proliferation and liver fibrosis in vivo in MT1
knockout (KO) mice, and wild-type (WT) mice in which the
hepatic expression of MT1 was reduced by Vivo-Morpholinos
for MT1. Methods: The studies were performed in normal (WT),
MT1 Morpholino-treated or MT1 KO mice with or without BDL
surgery. We evaluated the expression of MT1 in liver sections
by immunohistochemistry (IHC) and immunoblots in cholangiocytes.
Intrahepatic biliary ductal mass (IBDM) and biliary proliferation
was evaluated by IHC for CK-19 and PCNA in liver
sections, and qPCR for PCNA in cholangiocytes. Liver injury
was determined by H&E staining in liver sections and serum
levels of transaminases and bilirubin. Fibrosis was evaluated
by Sirius red staining in liver sections and qPCR for TGFbeta1,
alpha-SMA and fibronectin in cholangiocytes and total liver. In
vitro, shRNA-MT1 transfected murine cholangiocytes (MCCs)
and controls were used to measure proliferation and TGFbeta1,
alpha-SMA and fibronectin (FN-1) mRNA expression by qPCR.
Results: Normal cholangiocytes express low levels of MT1,
which markedly increased after BDL. We demonstrated that in
MT1 KO BDL and MT1 Morpholino-treated BDL mice, along
with reduced MT1 expression, there was decreased biliary
proliferation and IBDM. In MT1 KO BDL and MT1 Morpholino-treated
BDL mice, there was decreased lobular damage,
serum levels of transaminases and bilirubin, and liver fibrosis in
liver sections and reduced expression of TGFbeta1, alpha-SMA
and FN-1 compared to BDL WT mice. In shRNA-MT1 transfected
MCCs, there was decreased proliferation and reduced
expression of TGFb1, alpha-SMA and FN-1. The findings indicate
that shifting the melatonin axis to signal via MT2 during
the knockdown of MT1 expression decreases biliary proliferation
and liver fibrosis. Conclusion: Our findings indicate that
the balance of MT1/MT2 receptor expression for the melatonin
signaling axis may play an important role in the regulation of
biliary proliferation and hepatic fibrosis. Inhibition of MT1 may
be a key approach for ameliorating biliary hyperplasia and
liver fibrosis in cholestatic liver diseases.
Disclosures:
The following authors have nothing to disclose: Nan Wu, Fanyin Meng, Ying
Wan, Julie Venter, Holly A. Standeford, Heather L. Francis, Lindsey Kennedy,
Yuyan Han, Kelly McDaniel, Antonio Franchitto, Paolo Onori, Sharon DeMorrow,
Tami Annable, Eugenio Gaudio, Shannon S. Glaser, Gianfranco Alpini
836
Longer Term Efficacy of Simultaneous Liver Transplantation
plus Sleeve Gastrectomy versus non-invasive
Weight Loss followed by Liver Transplantation
Daniel Zamora-Valdes 1 , Kymberly D. Watt 2 , John J. Poterucha 2 ,
Charles B. Rosen 1 , Todd A. Kellogg 3 , Sara R. Di Cecco 2 , Nicki
M. Francisco Ziller 2 , Julie Heimbach 1 ; 1 Transplant Surgery, Mayo
Clinic, Rochester, MN; 2 Liver Transplant, Mayo Clinic, Rochester,
MN; 3 General Surgery, Mayo Clinic, Rochester, MN
Obesity is increasingly common before and after liver transplantation
(LT), yet optimal management remains unclear. Our
AIM was to analyze the long-term effectiveness of a multi-disciplinary
protocol for obese patients requiring LT, including
a non-invasive pre-transplant weight loss program, and combined
LT plus sleeve gastrectomy (SG) for obese patients who
failed to lose weight prior to LT. METHODS: since 2006, all
patients referred for LT with a BMI >35 were enrolled. There
are 46 patients who achieved weight loss and underwent LT
alone, and 17 who underwent LT combined with SG. Outcomes
for patients with > 2 years since LT alone or LT+SG
were analyzed using Student T-test for continuos variables and
c 2 for categorical variables. RESULTS: In those who received
LT alone, 34/46 were successful on achieving >10% loss in
total body weight (TBW) prior to LT. Of those with > 2 years
since LT, weight gain to BMI>35 was seen in 21/34 (61.7%)
at 2 years (Figure 1), though 10/34 (30%) who maintained
>10% loss in TBW at 2 years (p=0.005; Figure 1). There were
8/34 (23.5%) with diabetes post LT and 10/34 (18.9%) with
evidence of steatosis on year 2 ultrasound. In patients undergoing
the LT+SG, all patients achieved significant weight loss
(mean BMI at listing 49.8±1.5 vs. 2 year post LT 29.3±6.4;
p=0.001). No LT+SG patient developed post-LT DM or steatosis
at 2 years post LT. Conclusion: Non-invasive pre-transplant
weight loss was achieved by a majority, though weight gain
post LT was common. Combined LT plus SG resulted in effective
weight loss and was associated with fewer post-LT metabolic
complications at 2 years post LT.
Figure 1: post transplant BMI in morbidly obese patients who
underwent medical management (n=34) or sleeve gastrectomy
(n=5) combined with liver transplantation.
Disclosures:
The following authors have nothing to disclose: Daniel Zamora-Valdes, Kymberly
D. Watt, John J. Poterucha, Charles B. Rosen, Todd A. Kellogg, Sara R. Di
Cecco, Nicki M. Francisco Ziller, Julie Heimbach