In vitro biologic efficacy of sunitinib drug-eluting beads on human colorectal and hepatocellular carcinoma—A pilot study
RESEARCH ARTICLE
In vitro biologic efficacy of sunitinib drugeluting beads on human colorectal and
hepatocellular carcinoma—A pilot study
Steven Lahti1, Johannes M. Ludwig1,2, Minzhi Xing1,2, Lingyi Sun3, Dexing Zeng3*, Hyun
S. Kim1,2,4*
1 Interventional Oncology Translational Laboratory, Pittsburgh School of Medicine, Presbyterian South
Tower, Pittsburgh, PA, United States of America, 2 Division of Interventional Radiology, Department of
Radiology and Biomedical Imaging, Yale School of Medicine, New Haven, CT, United States of America,
3 Molecular Imaging Laboratory, Department of Radiology, University of Pittsburgh School of Medicine,
Pittsburgh, PA, United States of America, 4 Yale Cancer Center, Yale School of Medicine, New Haven, New
Haven, CT, United States of America
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* (HSK); (DZ)
Abstract
Purpose
OPEN ACCESS
Citation: Lahti S, Ludwig JM, Xing M, Sun L, Zeng
D, Kim HS (2017) In vitro biologic efficacy of
sunitinib drug-eluting beads on human colorectal
and hepatocellular carcinoma—A pilot study. PLoS
ONE 12(4): e0174539. https://doi.org/10.1371/
journal.pone.0174539
Editor: Yi-Hsien Hsieh, Institute of Biochemistry
and Biotechnology, TAIWAN
Received: April 30, 2016
Accepted: March 11, 2017
Published: April 6, 2017
Copyright: © 2017 Lahti et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited.
Data Availability Statement: All relevant data are
within the paper and its supporting information
files.
Funding: The authors received no specific funding
for this work.
Competing interests: The authors have declared
that no competing interests exist.
Sunitinib drug eluting beads (DEB) are a novel anti-angiogenic bead preparation for use in
transarterial chemoembolization. However, systematic studies of sunitinib DEB’s effect on
cancer cells have not been reported. Herein, we assess their direct biologic efficacy against
carcinoma cell lines and correlate cell viability with drug release in vitro.
Materials and methods
Sunitinib-HCl (10mg/mL) in Milli-Q water was mixed with LC Bead® 300–500μm (Biocompatibles UK Ltd.). Loading and release were assessed by measurement of drug UV absorbance using UV-visible spectrophotometer. Viability of human colorectal cancer (CRC,
HCT116 and HT29) and hepatocellular carcinoma (HCC, HepG2) cells upon exposure to
sunitinib DEB was measured using a bioluminescent assay. Drug concentration during
exposure was quantified using HPLC.
Results
When added to cultured HepG2 cells, sunitinib DEB rapidly inhibited viability with a significant decrease observed within 1 hour of incubation. Viability of HCT116 and HT29 cells
decreased relatively slower, with significant reductions observed after 8 and 24 hours,
respectively. After 24 hours there was nearly complete inhibition of all three cell lines. There
was no difference in viability observed between cells treated with 5 μl, 10 μL, or 20 μL of
sunitinib DEB. HPLC analysis of the cell culture supernatant demonstrated saturation of the
cell medium within approximately 4 hours for each amount added, with sunitinib achieving a
final concentration of 17.61 μM (SE ±1.01).
PLOS ONE | https://doi.org/10.1371/journal.pone.0174539 April 6, 2017
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Sunitinib drug-eluting bead for in vitro cancer treatment
Conclusions
Sunitinib can be efficiently loaded to and released from LC beads, and the resulting sunitinib
DEB demonstrate strong in vitro inhibition of human CRC and HCC cells.
Introduction
Transarterial chemoembolization (TACE) is a form of locoregional therapy that prolongs survival in patients with unresectable hepatocellular carcinoma (HCC), cholangiocarcinoma, and
a variety of hepatic metastases such as colorectal carcinoma [1]. The development of drug-eluting beads (DEB) as combined embolic agents and drug-delivery vehicles for use in TACE has
standardized treatment regimens, aiming at increasing intratumoral drug concentration and
objective tumor response, while decreasing systemic toxicity compared to the conventional
TACE based on the oily lipidol/drug emulsion [1,2]. However, the meta-analysis from Facciorusso et al. included 12 studies demonstrating equality between the two therapies in terms of
tumor response, survival and severe adverse events in patients with non-resectable HCC [2].
Moreover, a recent randomized trial (~50 patients per cohort) compared bland transarterial
embolization (TAE) vs. doxorubicin DEB-TACE demonstrating no survival differences in
treatment response, progression free survival, overall survival or adverse events rate [3].
Although no comparable data on the efficacy of Irinotecan loaded DEB in the treatment of
colorectal liver metastases are available [1,4], the data thus far compels further investigation of
other anticancer drugs that may be loaded onto DEB and upon release into the tumor, exert a
more potent antineoplastic effect than those already described.
One major limitation of DEB-TACE is its short-lived effect, which results in part because
DEB induced hypoxia stimulates neoangiogenesis and a recovery of tumor blood supply.
Numerous studies have documented increased expression of vascular endothelial growth factor (VEGF) in TACE treated tissues, as well as significantly increased serum/plasma levels of
VEGF thereafter [5–7]. Through its effects on neoangiogenesis and vascular permeability,
VEGF likely contributes to local recurrence and extrahepatic metastasis as has been investigated by previous studies [8–10].
Fuchs et al have recently synthesized an antiangiogenic DEB loaded with sunitinib and
have characterized the in vitro release characteristics, biologic efficacy against human umbilical
vein endothelial cells (HUVECs) and the in vivo release characteristics after TACE using a rabbit model [11]. In this study they demonstrated that beads loaded to 30 mg/g bead had a release
half-life of 1.1 and 1.6 hrs for 70–150 and 100–300 μm beads, respectively. They also observed
maximum in vitro release of 81 and 82% for these two bead sizes. Sunitinib DEB, furthermore,
were shown to inhibit HUVEC proliferation and migration in response to VEGF stimulation.
In a subsequent efficacy study [7], sunitinib loaded DEB achieved tumor growth arrest (-2%)
within 14 days of monitoring compared to oral sunitinib treatment (+1853%) and unloaded
DEB intra-arterial injection (+42%) (p = 0.044) of VX2 tumor implants in rabbits. Furthermore, a complete treatment response was observed in 60% of sunitinib loaded and in 43% of
unloaded DEB 14 days after treatment start.
Sunitinib (SU011248), a small molecule inhibitor of class III and class IV receptor tyrosine
kinases, including VEGFR1-3, PDGFR-α/β, c-Kit, Flt3, and RET, is currently approved for the
treatment of renal cell carcinoma (RCC) and imatinib-refractory gastrointest (...truncated)
