studies

916A AASLD ABSTRACTS HEPATOLOGY, October, 2015
1447
A novel canine liver cirrhosis model to develop less
invasive liver regeneration therapy using cultured bone
marrow-derived cells
Takashi Matsuda 1,2 , Taro Takami 1 , Yuki Aibe 1 , Toshihiko Matsumoto
1,4 , Tsuyoshi Ishikawa 1 , Naoki Yamamoto 1,5 , Shuji Terai 3 ,
Isao Sakaida 1,6 ; 1 Department of Gastroenterology and Hepatology,
Yamaguchi University Graduate School of Medicine, Ube,
Yamaguchi, Japan; 2 Sanyo-Onoda General Hospital, Sanyo-Onoda,
Japan; 3 Division of Gastroenterology and Hepatology,
Niigata University, Niigata, Japan; 4 Department of Oncology and
Laboratory Medicine, Yamaguchi University Graduate School of
Medicine, Ube, Yamaguchi, Japan; 5 Yamaguchi University Health
Administration Center, Yamaguchi University, Ube, Yamaguchi,
Japan; 6 Center for Reparative Medicine, Yamaguchi University
Graduate School of Medicine, Ube, Yamaguchi, Japan
Background/Aims: We developed autologous bone marrow
cell infusion (ABMi) therapy for decompensated liver cirrhosis
patients using non-cultured autologous whole bone marrow
(BM) cells aspirated under general anesthesia, and then
reported the safety and efficacy of this approach. We have
also been developing a new, less invasive liver regeneration
therapy using cultured autologous BM-derived mesenchymal
stem cells (BMSCs) from small amounts of BM fluid aspirated
under local anesthesia. Before human clinical trials, the safety
and efficacy of cultured autologous BMSC infusion in mediumto-large
animals must be confirmed; thus, we aimed to develop
a canine liver cirrhosis model. Methods: Eight 1- to 2-year-old
beagles were studied. A small amount of BM fluid was aspirated
from the canine humerus to assess the characteristics of
BMSCs. We implanted a human peripherally inserted central
venous catheter (PICC) in the stomach and a subcutaneous
infusion port in the back of the neck of each canine. Repeated
injection of carbon tetrachloride (CCl 4
) through the implanted
catheter (high-dose period: 0.75 mL/kg body weight, twice a
week for 6 weeks; low-dose period: 0.20 mL/kg body weight,
twice a week for 4 weeks) was performed to induce liver cirrhosis.
After 10 weeks of CCl 4
injection, eight canines were
equally divided into two groups: no cell infusion (control group)
and autologous BMSC infusion via the peripheral vein (BMSC
group). Low-dose CCl 4
was continued after BMSC infusion, and
blood examinations, ultrasonography-guided liver biopsies,
and indocyanine green (ICG) tests were carried out before and
at 4 weeks after the infusion. Results: Canine BMSCs cultured
in standard DMEM with 10% FBS adhered to plastic and were
CD44+, CD90+, and CD45−. They differentiated into adipocytes
and osteocytes, consistent with known characteristics
of BMSCs. In the BMSC group 4 weeks after BMSC infusion,
the area of Sirius red-stained liver fibrosis was significantly
reduced (fibrotic area (%); BMSCs: −1.62 ± 0.63; controls:
+1.44 ± 1.01, p < 0.05), consistent with a significantly shortened
half-life of ICG (half-life of ICG (min); BMSCs: −1.45 ±
0.42; controls: +1.23 ± 0.87, p < 0.05). After BMSC infusion,
no pro-coagulation activity and no thrombosis were seen in
lung arteries using contrast-enhanced computed tomography.
Conclusions: We established a useful canine liver cirrhosis
model with repeated CCl 4
administration through a PICC and
confirmed that cultured autologous BMSC infusion improved
liver cirrhosis and promoted liver regeneration without adverse
effects.
Disclosures:
The following authors have nothing to disclose: Takashi Matsuda, Taro Takami,
Yuki Aibe, Toshihiko Matsumoto, Tsuyoshi Ishikawa, Naoki Yamamoto, Shuji
Terai, Isao Sakaida
1448
A novel glycobiomarker, Wisteria floribunda agglutinin
Macrophage Colony-Stimulating Factor Receptor can
predict carcinogenesis and survival of liver cirrhosis
with hepatitis C virus
Etsuko Iio 1,3 , Makoto Ocho 2 , Akira Togayachi 2 , Masanori
Nojima 4 , Atsushi Kuno 2 , Masashi Mizokami 5 , Hiroshi Yatsuhashi 6 ,
Tatsuya Ide 7 , Shunsuke Nojiri 3 , Hisashi Narimatsu 2 , Yasuhito
Tanaka 1 ; 1 Virology and Liver Unit, Nagoya City University Graduate
School of Medical Sciences, Nagoya, Japan; 2 Research Center
for Medical Glycoscience, National Institute of Advanced Industrial
Science and Technology, Tsukuba, Japan; 3 Gastroenterology and
Metabolism, Nagoya City University Graduate School of Medical
Sciences, Nagoya, Japan; 4 Advanced Medicine Promotion,
The Advanced Clinical Research Center, The Institute of Medical
Science, University of Tokyo, Tokyo, Japan; 5 The Research Center
of Japan, Hepatitis and Immunology, Kohnodai Hospital, International
MedicalCenter, Ichikawa, Japan; 6 Clinical Research Center,
National Nagasaki Medical Center, Omura, Japan; 7 Division of
Gastroenterology, Department of Medicine, Kurume University,
Kurume, Japan
[Background/Aims] Recently, we identified a novel liver fibrosis
glycobiomarker Wisteria floribunda agglutinin (WFA)-reactive
colony stimulating factor 1 receptor (WFA + -CSF1R) using a glycoproteomics-based
strategy. The aim of the present study was
to assess the value of measuring WFA + -CSF1R levels for hepatocarcinogenesis
and outcome in liver cirrhosis (LC) patients
with hepatitis C virus (HCV). [Methods] WFA + -CSF1R and total
CSF1R levels were measured by an antibody-lectin sandwich
ELISA in serum samples from 214 consecutive HCV infected
patients (99 CH and 115 LC) from January 1998 to April 2013
in order to evaluate impact on carcinogenesis and survival of
LC patients. Among 115 patients with LC, 56 (48.7%) patients
without hepatocellular carcinoma (HCC). Ultimately, the 56 LC
patients and independent 45 LC patients without HCC were
assessed. [Results] LC patients without HCC of training cohort
(n = 56) and validation cohort (n = 45) at baseline were analyzed
for survival and carcinogenesis rates. The optimal cut-off
of the WFA + -CSF1R were determined to be 310 ng/ml according
to the smallest P value screened by Cox regression analysis
in the training set. The AUC of WFA + -CSF1R for predicting
overall survival, calculated by time-dependent ROC analysis,
was 0.691 and the HR (per 1-SD increase) was 1.80 (95% CI,
1.23-2.62, p < 0.001) of combined with the training set and
validation set. The survival rate of LC patients with high WFA + -
CSF1R levels (≥ 310 ng/ml) was significantly worse than those
with lower levels (p < 0.01). Next, we analyzed the percentage
of WFA + -CSF1R among total-CSF1R (WFA + -CSF1R%) for
the potential of HCC. The optimal cut-off for the WFA + -CSF1R%
for predicting the cumulative carcinogenesis rate was 35.0%.
We further verified the utility of WFA + -CSF1R%, using the validation
set. In the combined data the AUC of WFA + -CSF1R% for
predicting the cumulative carcinogenesis rate was 0.760, with
an HR of 1.66 (95% CI 1.26-2.20, p < 0.001). The AUC of
WFA + -CSF1R% was superior to other fibrosis and tumor markers
(i.e. Fib4, APRI, albumin, AFP, AFP-L3 and PIVKA-II) for predicting
the cumulative carcinogenesis rate. LC patients in the
training cohort with high WFA + -CSF1R% (≥ 35.0%, n = 6) had
cumulative carcinogenesis rates of 33.3%, 50.0%, and 75.0%
at 1, 3 and 5 years, respectively. In contrast, patients with
low WFA + -CSF1R% (< 35.0%, n = 50) had cumulative rates
of 4.6%, 18.0%, and 35.0%, respectively (p = 0.006). These
data were supported in the validation cohort. [Conclusions]
Assessing serum levels of WFA + -CSF1R has diagnostic value
for predicting carcinogenesis and the survival of LC patients.