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THE PANCASPASE INHIBITOR EMRICASAN IMPROVES THE PHENOTYPE OF HEPATOCYTES FROM HUMAN AND RAT CIRRHOTIC LIVERS WITHOUT EVIDENCE OF
HEPATOTOXICITY
Martí Ortega-Ribera1, Nicolò Manicardi1, Anabel Fernández-Iglesias1, Patricia Contreras2, Al Spada2, Jaume Bosch1 and Jordi Gracia-Sancho1*
1Liver Vascular Biology Research Group, Barcelona Hepatic Hemodynamic Laboratory, IDIBAPS Biomedical Research Institute - CIBERehd, University of Barcelona Medical School, Barcelona, Spain. 2Conatus Pharmaceuticals, San Diego, CA.
Chronic liver disease is characterized by extensive fibrosis, necroptosis and sinusoidal dysfunction1,2. The pancaspase inhibitor emricasan has shown promising results in improving sinusoidal microvascular dysfunction, liver fibrosis and portal hypertension in pre-clinical models of advanced chronic liver disease (Gracia-Sancho et al. AASLD16). Nevertheless, its effect on hepatocyte biology
have not been adequately studied. The aim of this study was to 1-elucidate the effect of emricasan in hepatocytes phenotype and function and 2-assess emricasan hepatotoxicity in chronically-injured hepatocytes using both in vitro and in vivo approches.
In vivo
We studied the effects of in vivo administration of emricasan on the phenotype and function of primary hepatocytes in cirrhotic rats (chronic CCl4) treated for 7 days with emricasan
(10mg/kg/day) or vehicle (carboxymethylcellulose).
In vitro
We assessed the effects of 24h-emricasan (10-50 μM), or vehicle treatment, on the phenotype of primary human hepatocytes isolated from cirrhotic liver explants (EtOH
aetiology) cultured in an advanced fluidic device that mimics the liver sinusoid3,4.
Diclofenac was used as a positive control for hepatotoxicity.
Human cirrhotic hepatocytes treated with emricasan exhibited improved phenotype as demonstrated by higher albumin and urea
production.
Hepatocytes isolated from emricasan-treated cirrhotic rats showed increased albumin and urea production and superior CYP3A4 activity suggesting improved synthetic and detoxification capacity. Moreover, hepatocyte phenotype was maintained as evidenced by sustained
expression of the master regulator Hnf4α and Abcc3, Slc22a7, Slc22a1, Slc10a1 and Abcc2 transporters.
Emricasan showed no hepatotoxicity neither in human nor rat hepatocytes shown as the maintenance of cell viability and normal levels of transaminases and LDH in cell media (advanced fluidic device in vitro) and improved biochemical tests in emricasan-treated rats (in vivo).
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CYP4503A4Activity
RLU
(re
lati
ve t
o v
eh
icle
)
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1
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Urea Albumin
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rmal
ize
d u
g/h
*10
^6
he
pat
ocy
tes
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Hnf4α Abcc3 Slc22a1 Abcc2 Slc10a1 Slc22a7 Abcc4 Slco1a2
mR
NA
exp
ress
ion
re
lati
ve t
o
Ve
hic
le
*Corresponding author’s email: [email protected]
M.O.-R. has an i-PFIS fellowship from Instituto de Salud Carlos III (ISCIII). This study was funded by Conatus Pharmaceuticals.
@HH_lab
1- Fernández-Iglesias A. et al. (2017) Frontiers in medicine
2- Marrone G. et al. (2016) Journal of Hepatology
3- Illa X. et al. (2014) PlosOne
4- Ortega-Ribera M. et al. (2016) EASL
Emricasan improves the phenotype of cirrhotic human and rat hepatocytes without evidence of hepatotoxicity, further encouraging its clinical evaluation for the treatment of chronic liver disease.
METHODOLOGY
RESULTS
CONCLUSIONS REFERENCES
Vehicle Emricasan
0 μM 50 μM 10 μM
BACKGROUND AND AIMS
CONTACT AND FUNDING
Vehicle Emricasan
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