studies

410A AASLD ABSTRACTS HEPATOLOGY, October, 2015
Disclosures:
Vahakn Shahinian - Consulting: Amgen, Inc
The following authors have nothing to disclose: Neehar D. Parikh, Vincent D.
Marshall, Hari Nathan, Rajesh Balkrishnan
396
The Casein kinase II (CK2) inhibitor CX-4945 has
an additive effect with gemcitabine and cisplatin on
cholangiocarcinoma (CCA) cell lines mediated in part
through PI3K/Akt inhibition
Kais Zakharia 1 , Katsuyuki Miyabe 1 , Catherine D. Moser 1 , Sean
O’Brien 3 , Mitesh J. Borad 2 , Lewis R. Roberts 1 ; 1 Division of Gastroenterology
and Hepatology, Mayo Clinic College of Medicine,
and Mayo Clinic Cancer Center, Rochester, MN; 2 Division of
Hematology, Division of Oncology, Mayo Clinic, Scottsdale, AZ;
3 Senhwa Biosciences, San Diego, CA
Background: There is a need to develop effective therapies for
CCA due to suboptimal outcomes with the current standard
gemcitabine (Gem) and cisplatin (Cis) therapy. Casein kinase
II (CK2) is a serine/threonine selective protein kinase that has
been implicated in cancer progression, making it an attractive
target for anticancer treatment. CX-4945, a first in class, orally
available and highly selective inhibitor of CK2 has shown antitumor
activity in other human cancers. In this study we investigated
the anti-neoplastic effect of CX-4945 on 3 different
CCA cell lines. Aims: To determine 1) whether CX-4945 has
an anti-proliferative effect on CCA cells; 2) if CX-4945 induces
apoptosis in CCA cells; 3) whether the anti-proliferative action
of CX-4945 occurs through effects on PI3K/Akt signaling; and
4) whether the effect of CX-4945 enhances that of Gem or Cis
in vitro. Methods: CCK8 and colony formation assays were
used to assess cell viability in cultures of HuCCT1 (intrahepatic
CCA), EGI-1 (extrahepatic CCA), and a new primary patient
derived CCA cell line LIV27 after treatment with CX-4945
alone or in combination with Gem or Cis. The Caspase 3/7
assay was used to assess the effect of CX-4945 on CCA apoptosis.
The half maximal inhibitory concentrations (IC50) were
calculated using the Chou-Talalay technique. Western immunoblotting
was used to assess the effect of CX-4945 on Akt
pathway signaling in CCA cells. Results: CX-4945 significantly
decreased cell viability of HuCCT1, EGI-1 and LIV27 in a timeand
dose-dependent manner (IC50 of 7.3, 9.5, and 9.4 μM
respectively at 72 hours). CX-4945 inhibited colony formation
in HuCCT1 and EGI-1 (IC50 of 3.8 and 1.6 μM respectively).
CX-4945 did not significantly increase Caspase 3/7 activity
in CCA cells until higher doses (15 and 20 μM) were used.
CX-4945 inhibited Akt phosphorylation (at Serine473 and
Serine129) and PTEN phosphorylation, suggesting that it acts
through the PI3K/Akt pathway. Addition of CX-4945 to Cis
or Gem decreased viability of the HuCCT1, EGI-1 and LIV27
cells by 15-40% (depending on the dose and the cell line)
with a combination index (CI) of approximately 1, indicating
an additive effect. Addition of CX-4945 to both Gem and Cis
together (1/4 IC50s) decreased viability of HuCCT1 cells by
10-30%, also with a CI around 1. Conclusion: CX-4945 has an
anti-proliferative effect on CCA cell lines. Only higher doses of
CX-4945 increased Caspase 3/7 activity. The anti-proliferative
effect of CX-4945 correlates with effects on PI3K/Akt signaling.
CX-4945 enhanced the anti-proliferative effect of both Cis and
Gem in vitro. An animal study is being performed to confirm
these findings in vivo.
Disclosures:
Sean O’Brien - Employment: Senhwa Biosciences
Lewis R. Roberts - Grant/Research Support: Bristol Myers Squibb, ARIAD Pharmaceuticals,
BTG, Wako Diagnostics, Inova Diagnostics, Gilead Sciences, Five
Prime Therapeutics
The following authors have nothing to disclose: Kais Zakharia, Katsuyuki Miyabe,
Catherine D. Moser, Mitesh J. Borad
397
Aspirin Use is Associated with Reduced Risk of Cholangiocarcinoma
Jonggi Choi 1 , Hassan M. Ghoz 1 , Thoetchai Peeraphatdit 1,2 , Esha
Baichoo 1,3 , Benyam D. Addissie 1 , William S. Harmsen 4 , Terry M.
Therneau 4 , Janet E. Olson 4,5 , Roongruedee Chaiteerakij 1,6 , Lewis
R. Roberts 1 ; 1 Division of Gastroenterology and Hepatology, Mayo
Clinic College of Medicine, and Mayo Clinic Cancer Center, Rochester,
MN; 2 Department of Internal Medicine, University of Minnesota,
Minneapolis, MN; 3 Department of Medicine, Mount Sinai
Roosevelt Hospital, New York, NY; 4 Department of Biomedical
Statistics and Informatics, Mayo Clinic, Rochester, MN; 5 Division
of Epidemiology, Department of Health Sciences Research, Mayo
Clinic, Rochester, MN; 6 Department of Medicine, Faculty of Medicine,
Chulalongkorn University and King Chulalongkorn Memorial
Hospital, Thai Red Cross Society, Bangkok, Thailand
Background & Aims: It is unclear whether aspirin reduces cholangiocarcinoma
(CCA) risk. Previous studies on risk factors
for CCA combined all the three CCA subtypes (intrahepatic,
perihilar, and distal) together or combined perihilar and distal
CCA as extrahepatic CCA. Whether these factors conferred
the risk of all 3 subtypes to a similar magnitude is unknown.
We aimed to determine the associations between aspirin use
and other risk factors for each CCA subtype individually. Methods:
We enrolled 2,395 CCA cases (1,169 intrahepatic, 995
perihilar and 231 distal) seen at Mayo Clinic, Rochester, MN
between 2000 and 2014. Controls selected from the Mayo
Clinic Biobank were matched 2:1 with cases by age, sex,
race, and residential area (n=4,769). Associations between
aspirin use and other factors and CCA risk were determined
using logistic regression analysis. Results: There were 591
(24.7%) CCA cases and 2,129 (44.6%) controls who took
aspirin. Of aspirin users, 560 (94.8%) of cases and 2008
(94.3%) of controls were daily users; 384 (65.0%) of cases
and 1,697 (79.7%) of controls took low-dose aspirin (81-162
mg/day). Aspirin use had a significant inverse association
for all CCA subtypes with adjusted odds ratios (AOR) (95%
confidence interval, 95% CI) of 0.35 (0.29-0.42), 0.34 (0.27-
0.42), and 0.29 (0.19-0.44), for intrahepatic, perihilar and
distal CCA, respectively, p