studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 499A
583
Phenotypic and genotypic characterization of drug-induced
liver injury (DILI) with autoimmune features
C. Stephens 1 , A. Ortega 1 , I. Medina-Cáliz 1 , Mercedes Robles
Diaz 1 , Agustin Castiella 2 , Pedro Otazua 3 , E. Zapata 2 , E.M.
Gomez-Moreno 4 , MAngel López-Nevot 4 , Francisco Ruiz-Cabello 4 ,
German Soriano 5 , E. Roman 5,6 , Hacibe Hallal 7 , Jose María
Moreno-Planas 8 , Martin Prieto 9 , M. I. Lucena 1 , Raul J. Andrade 1 ;
1 Instituto de Investigación Biomédica de Málaga (IBIMA), Hospital
Universitario Virgen de la Victoria, Universidad de Málaga,
CIBERehd, Málaga, Spain; 2 Hospital Mendaro, Guipúzcoa, Spain;
3 Hospital Mondragón, Guipúzcoa, Spain; 4 Instituto de Investigación
Biosanitario de Granada, Hospital Universitario Virgen de
las Nieves, Universidad de Granada, Granada, Spain; 5 Hospital
de la Santa Creu i Sant Pau, Universitat Autònoma de Barcelona,
CIBERehd, Barcelona, Spain; 6 Escola Universitària d’Infermeria
EUI-Sant Pau, Universitat Autònoma de Barcelona, Barcelona,
Spain; 7 Hospital Morales Meseguer, Murcia, Spain; 8 Complejo
Hospitalario Universitario de Albacete, Albacete, Spain; 9 Hospital
La Fe, CIBERehd, Valencia, Spain
Background: Positive autoantibody (AAB) titres is a common
presentation of autoimmune hepatitis (AIH), but can also occur
in DILI. The underlying mechanism of selective AAB occurrence
in DILI is unknown. Hence, we aimed to compare demographics,
clinical parameters and HLA allele compositions in DILI
with positive (+) and negative (-) AAB titres and AIH patients.
Material and Methods: High resolution genotyping of HLA
class I (A, B, C) and II (DRB1, DQB1) loci were performed
on 207 DILI (drug-induced autoimmune hepatitis excluded)
and 51 AIH patients and compared to 885 Spanish healthy
controls. Results: Fifty-eight of the 207 DILI patients presented
positive titres for at least one AAB (ANA 76%, ASMA 28%,
AMA 9% or LKM-1 2%) during the DILI episode, while 149
were negative for all four AABs. Comparing demographics
and clinical parameters, AAB+ patients were found to be significantly
older than AAB- patients (58 vs 50 years, p=0.019).
Females predominated among the AIH (71%) compared to
AAB+ (59%) and AAB- (47%), p=0.01. Significant differences
in type of liver injury were present (p=0.0025), with cholestatic/mixed
damage being more prevalent in AAB-, which was
reflected in a higher onset ALP elevation in the same group
(p=0.029). Furthermore, hypertension was significantly more
frequent in AAB+ patients. Compared to controls, HLA alleles
B*08:01 (45% vs 10%, pc=1.0E-12), C*07:01 (47% vs 24%,
pc=0.006), DRB1*03:01(59% vs 26%, pc=2.0E-05) and
DQB1*02:01 (57% vs 22%, pc=3.0E-07) were significantly
more frequent in AIH patients. The frequency of HLA-A*01:01
was increased in the same population, but did not reach significance
after Bonferroni’s correction (33% vs 19%, p=0.02/
pc=0.37). There was a tendency for higher representation of
DRB1*14:01 and DQB1*05:03 in DILI AAB+ compared to
DILI AAB- (14% vs 4.7%, p=0.03; 14% vs 5.4%, p=0.06) and
controls (14% vs 5.0%, p=0.007; 14% vs 5.4%, p=0.01).
Conclusions: The AAB+ patients were found to be older and
less prevalent to develop cholestatic/mixed injury compared
to AAB- patients. The AIH patients were predominately female
with hepatocellular injury and less likely to have underlying
hypertensive conditions. The presence of HLA alleles B*08:01,
C*07:01, DRB1*03:01 and DQB1*02:01 and possibly
A*01:01 appear to enhance the risk of AIH in Caucasians with
Spanish inheritance. These alleles form part of the conserved
extended haplotype 8.1. However, haplotype formations in the
study cohort are currently unknown. HLA alleles DRB1*14:01
and DQB1*05:03 could potentially increase the risk of positive
AAB (particularly ANA) in Spanish DILI patients. Funding:
P10-CTS-6470, PI12/00378, AC-0073-2013, CIBERehd-ISCIII
Disclosures:
Martin Prieto - Advisory Committees or Review Panels: Bristol, Gilead
The following authors have nothing to disclose: C. Stephens, A. Ortega, I. Medina-Cáliz,
Mercedes Robles Diaz, Agustin Castiella, Pedro Otazua, E. Zapata,
E.M. Gomez-Moreno, MAngel López-Nevot, Francisco Ruiz-Cabello, German
Soriano, E. Roman, Hacibe Hallal, Jose María Moreno-Planas, M. I. Lucena,
Raul J. Andrade
584
Effect of Branched Chain Amino Acids on Iron Transport
in the Liver and Small Intestine of a Cirrhotic Rat Model
Yoshinao Kobayashi 1,2 , Ryosuke Sugimoto 2 , Motoh Iwasa 2 , Yoshiyuki
Takei 2 ; 1 Center for Physical and Mental Health, Mie University
Graduate School of Medicine, Tsu, Japan; 2 Department of Gastroenterology
and Hepatology, Mie University Graduate School of
Medicine, Tsu, Japan
BACKGROUND: Branched chain amino acids (BCAA) have
been used as a supplemental therapy to improve malnutrition
and event-free survival in patients with liver cirrhosis (LC). Iron
is known to involve in inflammation and carcinogenesis of the
liver through production of reactive oxygen spices (ROS). We
have reported that BCAA reduce oxidative stress by diminished
hepatic iron accumulation in a cirrhotic rat model. In order to
clarify the effect of BCAA supplementation on iron metabolism
of the liver and small intestine, we investigated effects of BCAA
on hepatic iron accumulation and intestinal iron transporters
in a rat model. Additionally, we investigated effects of BCAA
on expression of intestinal iron transporters using CaCo2 cells.
METHODS: Carbon tetrachloride (CCl 4
) was administered to
male Wistar rats. On the 5 th week, rats were randomly divided
into 3 groups and further maintained for 16 weeks. Group 1
(G1) was treated with basal diet as a control (n=8). Group 2
was treated with non-heme iron diet (60mg/kg a day) (n=7).
Group 3 was treated both with non-heme iron and BCAA
mixture (10mg/kg a day) (n=7). On the 21 st week, liver and
intestine tissues were collected. Expression of iron transporters,
divalent metal transporter 1 (Dmt1) and ferroportin 1 (Fp1),
was analyzed by a semi-quantitative RT-PCR and Western blotting.
Furthermore, Caco2 cells were cultured for 48 hours with
FeCl 3
, deferoxamine (DFO) and/or BCAA. RESULTS: We have
shown that BCAA-supplemented rats showed lower hepatic iron
contents (P< 0.05), lower hepatic hepcidin mRNA expression
(P< 0.001), lower serum hepcidin level (P= 0.05) and lower
hepatic 8-hydroxyl-2’-deoxyguanosine (8-OHdG) production
(P< 0.05), as compared with control rats. Protein expression
of Dmt1 in the small intestine was reduced in G2 (vs. G1, P <
0.05), which tended to be retrieved in G3 (vs. G2, P= 0.08).
Protein expression of intestinal Fp1 tended to be reduced in G2
(vs. G1, P=0.06). The reduced FP1 tended to be retrieved in
G3 (vs. G2, P= 0.09). BCAA did not affect gene and protein
expression of Dmt1 in Caco2 cells. Gene expression of Fp1 in
these cells was not significantly changed by BCAA, whereas
protein expression of Fp1 in the cell membrane fraction was
reduced when cultured with BCAA. Among BCAA, leucine
most strongly repressed FP1 protein expression in Caco2 cells.
CONCLUSION: BCAA, especially leucine, can reduce intestinal
Fp1 protein expression through post-transcriptional pathway,
which was a possible mechanism for reducing intestinal iron
absorption. BCAA and serum hepcidin level reduced by BCAA
supplementation can modulate iron absorption from the intestine
and hepatic iron accumulation.
Disclosures:
The following authors have nothing to disclose: Yoshinao Kobayashi, Ryosuke
Sugimoto, Motoh Iwasa, Yoshiyuki Takei