studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 1151A
1933
Genome-wide investigation identifies common SNPs on
Chr9p22.2 as risk factors for liver injury due to sulfamethoxazole/trimethoprim
Thomas J. Urban 1,2 , Paola Nicoletti 3 , Robert J. Fontana 4 , Andrew
Stolz 5 , Ann K. Daly 6 , Guruprasad P. Aithal 7 , M. I. Lucena 8 , Huiman
X. Barnhart 9 , Paul Watkins 2 , Naga P. Chalasani 10 ; 1 Eshelman
School of Pharmacy, University of North Carolina at Chapel
Hill, Chapel Hill, NC; 2 Hamner Institute for Drug Safety Sciences,
Durham, NC; 3 Columbia University, New York, NY; 4 University
of Michigan, Ann Arbor, MI; 5 University of Southern California,
Los Angeles, CA; 6 University of Newcastle, Newcastle upon Tyne,
United Kingdom; 7 Nottingham University, Nottingham, United
Kingdom; 8 University of Malaga, Malaga, Spain; 9 Duke University,
Durham, NC; 10 Indiana University School of Medicine, Indianapolis,
IN
BACKGROUND &AIM: Treatment with the antibiotic combination
sulfamethoxazole/trimethoprim (SMZ/TMP) can result in a
rare but serious liver toxicity which typically presents with immunoallergic
features. Currently there are no genetic or non-genetic
predictors of toxicity. We aimed to uncover genetic risk
factors associated with SMZ/TMP DILI. METHODS: Genomewide
genotyping was performed on 27 individuals of European
ancestry who experienced liver injury from SMZ/TMP, including
22 DILIN patients and 5 from iDILIC/iSAEC. Genotype
frequencies were compared to >5,000 population controls.
Targeted genotyping by TaqMan assay was performed for validation
in 33 cases (19 cases included in the GWAS and 14
new cases not included in the GWAS). RESULTS: GWAS was
conducted on 27 individuals with SMZ/TMP DILI with mean
age 47 years, 48% females, and the pattern of liver injury was
hepatocellular in 41% and cholestatic in 30%. Their peak ALT
(mean ± sd) were 714 ± 542 U/L and peak bilirubin 11.5±
10.1 mg/dL. The GWAS revealed a set of common SNPs in an
intergenic region on Chr9p22.2 that showed strong evidence
of association with SMZ/TMP-DILI. The minor allele frequency
(MAF) of the risk allele was 26% in SMZ/TMP-DILI cases, vs.
4.6% in controls (OR: 9.0, 95%CI: 4.3-19.1, p = 5 X 10 -9 ).
Validation cohort consisted of 33 European Caucasians (19
from the GWAS plus 14 independent replication samples) with
mean age 46 years, 55% females and the pattern of liver injury
was hepatocellular in 28% and cholestatic in 32%. Their peak
ALT values were 745 ± 695 U/L and bilirubin 13.1± 10.1 mg/
dL. Targeted assays confirmed the genotypes as measured by
the GWAS arrays, and further identified 4 additional carriers
among 14 new cases in the validation cohort (MAF=14.3%,
p < 0.05). However, the phenotypic characteristics of DILI in
patients who carried the risk alleles (age, sex, latency to onset,
immunoallergic features, peak laboratory test values) were not
significantly different from those who lacked the risk alleles at
Chr9p22.2. CONCLUSIONS: Common SNPs on Chr9p22.2
are strongly associated with risk for SMX/TMP-DILI. These
SNPs are not located near any known protein-coding gene or
miRNA, and have not previously been associated with mRNA
expression of any gene. Further studies will be necessary to
understand the mechanism underlying the association, and to
extend these studies to other hypersensitivity reactions to SMZ/
TMP as well as to DILI caused by other sulfonamides.
Disclosures:
Robert J. Fontana - Consulting: GlaxoSmithKline, CLDF; Grant/Research Support:
Gilead, vertex, BMS, Jansen, Gilead
Naga P. Chalasani - Consulting: Abbvie, Lilly, Celgene, Tobira, NuSirt, Takeda,
Merck/Anthem, Salix; Grant/Research Support: Intercept, Gilead, Galectin
The following authors have nothing to disclose: Thomas J. Urban, Paola Nicoletti,
Andrew Stolz, Ann K. Daly, Guruprasad P. Aithal, M. I. Lucena, Huiman X.
Barnhart, Paul Watkins
1934
Co-activation of AKT and c-Met triggers rapid hepatocellular
carcinoma development via mTORC1/FASN
pathway in mice
Junjie Hu 2,1 , Lei Li 3,1 , Li Che 1 , Diego Calvisi 4 , Xin Chen 1 ; 1 Bioengineering
and Therapeutic Sciences, UCSF, San Francisoco,
CA; 2 School of Pharmacy, Hubei University of Chinese Medicine,
Wuhan, China; 3 School of Pharmacy, Huazhong university of science
and technology, Wuhan, China; 4 Institude of Pathology, university
of greifswald, Greifswald, Germany
Background: Activation of the AKT/mTOR cascade and overexpression
of c-Met have been implicated in the development
of human hepatocellular carcinoma (HCC). Methods: To elucidate
the functional crosstalk between the AKT and c-Met pathways,
we generated a model characterized by the combined
expression of activated AKT and c-Met in the mouse liver using
hydrodynamic transfection. Results: We found that co-expression
of AKT and c-Met results in faster liver tumor development
when compared with mice overexpressing AKT alone, whereas
c-Met alone does not lead to tumor formation. At the molecular
level, liver tumors induced by AKT/c-Met display activation of
AKT/mTOR and Ras/MAPK cascades as well as increased
lipogenesis and glycolysis. Mechanistically, AKT/c-Met driven
hepatocarcinogenesis was found to be mTORC1-dependent, as
Raptor genetic deletion abrogates tumor development in AKT/
c-Met mice. Furthermore, since increased de novo synthesis of
fatty acid is a pivotal metabolic event downstream of mTORC1,
we determined the requirement of lipogenesis in AKT/c-Met
driven liver tumor development using conditional Fatty Acid
Synthase (FASN) knockout mice. Of note, hepatocarcinogenesis
induced by AKT/c-Met was fully inhibited by FASN ablation.
In human HCC samples, coordinated expression of FASN,
c-Met, activated AKT and ERK proteins was detected in a subgroup
of biologically aggressive tumors. Conclutions: Our study
demonstrates that co-activation of AKT and c-Met induces HCC
development that depends on the mTORC1/FASN pathway.
Suppression of mTORC1 and/or FASN might be highly detrimental
for the growth of human HCC subsets characterized by
concomitant induction of the AKT and c-Met cascades.
Disclosures:
The following authors have nothing to disclose: Junjie Hu, Lei Li, Li Che, Diego
Calvisi, Xin Chen
1935
Classification of tumors based on the integrated profile
of genetic and epigenetic alterations and the biological
behavior of human hepatocellular carcinoma
Naoshi Nishida 1 , Toshimi Kaido 2 , Masatoshi Kudo 1 ; 1 Department
of Gastroenterology and Hepatology, Kinki University School of
Medicine, Osaka-sayama, Japan; 2 Department of Surgery, Graduate
School of Medicine Kyoto University, Kyoto, Japan
Aims: We tried to classify HCCs based on the integrated
genetic and epigenetic profile, and examined the association
between the classification and biological characteristics
such as high metastatic potential. Methods: (1) A total of 121
HCCs were examined for p53, β-catenin and TERT promoter
mutation, methylation of 8 tumor suppressor genes (TSGs)
and hypomethylation on 3 repetitive DNA sequences (RSs),
and fractional allelic loss (FAL) as a surrogate marker of chromosomal
instability (CI) using direct sequencing, MethyLight,
and microsatellite analyses with 200 markers, respectively. A
degree of TSG methylation and hypomethylation of RSs was
categorized as “extensive” vs. “limited” and “significant” vs.
“slight” by hierarchical clustering, respectively. Each tumor was