ImmunoPET as Stoichiometric Sensor for Glypican-3 in Models of Hepatocellular Carcinoma

BackgroundHepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. While conventional imaging approaches like ultrasound, CT, and MRI play critical roles in the diagnosis and surveillance of HCC, improved methods for detection and assessment of treatment response are needed. One promising approach is the use of radiolabeled antibodies for positron emission tomography (immunoPET) imaging. Glypican-3 (GPC3) is a proteoglycan that is highly expressed in the majority of HCC tumors. GPC3-specific antibodies are used to diagnose HCC histopathologically, and have been proposed as a treatment of HCC. Here, we design, synthesize and demonstrate that our humanized immunoPET agent, [89Zr]Zr-DFO-TAB-H14, can stoichiometrically bind to models of human liver cancer with varied GPC3 expression. Methods: The GPC3-specific monoclonal humanized IgG1, TAB-H14, was used as a scaffold for engineering our immunoPET agent. Fluorescent and deferroxamine (DFO) chelate conjugates of TAB-H14 were characterized using mass spectrometry. Binding affinity of TAB-H14 and conjugates for GPC3 was determined in cell-free biolayer interferometry, and cell-based radioimmunoassays. GPC3-expression was assessed by flow cytometry and immunofluorescence using commercially available anti-GPC3 antibodies and TAB-H14 in GPC3−(A431) and GPC3+ cell lines including an engineered line (A431-GPC3+, G1) and liver cancer lines (HepG2, Hep3B, and Huh7). DFO-TAB-H14, was radiolabeled with Zr-89. Mice were subcutaneously engrafted with the aforementioned cell lines and in vivo target engagement of the immunoPET agent [89Zr]Zr-DFO-TAB-H14 was determined using PET/CT, quantitative biodistribution, and autoradiography. Results: TAB-H14 demonstrated subnanomolar to nanomolar affinity for human GPC3. Fluorescently tagged TAB-H14 was able to bind to GPC3 on cell membranes of GPC3-expressing lines by flow cytometry. These results were confirmed by immunofluorescence staining of A431, G1 HepG2, Hep3B, and Huh7 tumor sections. ImmunoPET imaging with [89Zr]Zr-DFO-TAB-H14 showed stoichiometric tumor uptake corresponding to the cell surface expression levels. Autoradiography and immunostaining confirmed in vivo findings. Conclusion: We systematically demonstrate that the humanized immnoPET agent [89Zr]Zr-DFO-TAB-H14 specifically and stoichiometrically binds to GPC3 in several models of human liver cancer, serving as a promising in vivo GPC3 sensor. This agent may provide utility in HCC diagnosis and surveillance, and the selection of candidates for GPC3-directed therapies.

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Potential Role of CD133 Expression in the Susceptibility of Human Liver Cancer Stem-Like Cells to TRAIL

Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics ◽

10.3727/096504016x14685034103950 ◽

2016 ◽

Vol 24 (6) ◽

pp. 495-509 ◽

Cited By ~ 9

Author(s):

Su-Hoon Lee ◽

Suh-Kyung Hyun ◽

Hak-Bong Kim ◽

Chi-Dug Kang ◽

Sun-Hee Kim

Keyword(s):

Liver Cancer ◽

Cell Lines ◽

Human Liver ◽

Surface Expression ◽

Small Population ◽

Cell Surface Expression ◽

High Recurrence Rate ◽

Cd133 Expression ◽

New Strategy ◽

Human Liver Cancer

Hepatocellular carcinoma (HCC) is one of the most common malignancies, with a poor prognosis and high recurrence rate. In the present study, we identified CD133, one of the markers of cancer stem cells, as a novel molecular target of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). In four human HCC cell lines established from primary HCC tumors, we found that CD133-high human liver cancer stem-like cells (CD133hi) derived from the SNU-475 cell line were highly susceptible to TRAIL compared to other HCC cell lines with a small population of CD133. CD133hi SNU-475 cells showed upregulation of TRAIL receptor DR5 and stemness-related genes such as c-Myc and ABC transporters compared to their CD133-low (CD133lo) cells. Hypersensitivity of CD133hi cells to TRAIL was associated with c-Myc-mediated upregulation of DR5 and downregulation of c-FLIPL in the cells. Knockdown of CD133 expression in CD133hi cells resulted in the downregulation of c-Myc, and depletion of c-Myc caused a decrease in the cell surface expression of DR5 and an increase in the expression of c-FLIPL and, consequently, attenuated TRAIL-induced cytotoxicity and apoptosis of CD133hi cells. These results suggest that TRAIL may provide a new strategy for CD133hi CSCs of HCC-targeted therapies and, potentially, for therapies of other CD133-expressing types of cancer.

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Antiproliferative Effect of Lenvatinib on Human Liver Cancer Cell Lines In Vitro and In Vivo

Anticancer Research ◽

10.21873/anticanres.13802 ◽

2019 ◽

Vol 39 (11) ◽

pp. 5973-5982 ◽

Cited By ~ 3

Author(s):

SACHIKO OGASAWARA ◽

YUTARO MIHARA ◽

REIICHIRO KONDO ◽

HIRONORI KUSANO ◽

JUN AKIBA ◽

...

Keyword(s):

Liver Cancer ◽

Cell Lines ◽

Cancer Cell ◽

Human Liver ◽

Antiproliferative Effect ◽

Liver Cancer Cell ◽

Human Liver Cancer Cell ◽

Human Liver Cancer

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A Titanium (IV)–Dithiophenolate Complex and Its Chitosan Nanocomposite: Their Roles towards Rat Liver Injuries In Vivo and against Human Liver Cancer Cell Lines

International Journal of Molecular Sciences ◽

10.3390/ijms222011219 ◽

2021 ◽

Vol 22 (20) ◽

pp. 11219

Author(s):

Nadia Z. Shaban ◽

Salah A. Yehia ◽

Doaa Awad ◽

Shaban Y. Shaban ◽

Samar R. Saleh

Keyword(s):

Oxidative Stress ◽

Liver Cancer ◽

Cell Lines ◽

Rat Liver ◽

Liver Damage ◽

Human Liver ◽

Therapeutic Effects ◽

Liver Injuries ◽

Human Liver Cancer

Titanium (IV)–dithiophenolate complex chitosan nanocomposites (DBT–CSNPs) are featured by their antibacterial activities, cytotoxicity, and capacity to bind with DNA helixes. In this study, their therapeutic effects against rat liver damage induced by carbon tetrachloride (CCl4) and their anti-proliferative activity against human liver cancer (HepG2) cell lines were determined. Results of treatment were compared with cisplatin treatment. Markers of apoptosis, oxidative stress, liver functions, and liver histopathology were determined. The results showed that DBT–CSNPs and DBT treatments abolished liver damage induced by CCl4 and improved liver architecture and functions. DNA fragmentation, Bax, and caspase-8 were reduced, but Bcl-2 and the Bcl-2/Bax ratios were increased. However, there was a non-significant change in the oxidative stress markers. DBT–CSNPs and DBT inhibited the proliferation of HepG2 cells by arresting cells in the G2/M phase and inducing cell death. DBT–CSNPs were more efficient than DBT. Low doses of DBT and DBT–CSNPs applied to healthy rats for 14 days had no adverse effect. DBT and DBT–CSNP treatment gave preferable results than the treatment with cisplatin. In conclusion, DBT–CSNPs and DBT have anti-apoptotic activities against liver injuries and have anti-neoplastic impacts. DBT–CSNPs are more efficient. Both compounds can be used in pharmacological fields.

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Modulation by cytokines on the surface expression and shedding of intercellular adhesion molecule-1 (ICAM-1) in human liver cancer cell lines

International Hepatology Communications ◽

10.1016/0928-4346(94)90011-6 ◽

1994 ◽

Vol 2 (1) ◽

pp. 56-61 ◽

Cited By ~ 3

Author(s):

Y SHIMIZU ◽

K OKADA ◽

T TSUKISHIRO ◽

M MINEMURA ◽

K HIGUCHI ◽

...

Keyword(s):

Liver Cancer ◽

Cell Lines ◽

Adhesion Molecule ◽

Human Liver ◽

Surface Expression ◽

Intercellular Adhesion Molecule ◽

Liver Cancer Cell ◽

Intercellular Adhesion Molecule 1 ◽

Human Liver Cancer Cell ◽

Human Liver Cancer

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TGF-β inactivation and TGF-α overexpression cooperate in an in vivo mouse model to induce hepatocellular carcinoma that recapitulates molecular features of human liver cancer

International Journal of Cancer ◽

10.1002/ijc.25127 ◽

2009 ◽

Vol 127 (5) ◽

pp. 1060-1071 ◽

Cited By ~ 15

Author(s):

Ji Yeon Baek ◽

Shelli M. Morris ◽

Jean Campbell ◽

Nelson Fausto ◽

Matthew M. Yeh ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Cancer ◽

Mouse Model ◽

Human Liver ◽

Molecular Features ◽

Human Liver Cancer

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EZH2-mediated epigenetic silencing of ZIC4 in hepatocellular carcinoma

10.21203/rs.3.rs-30023/v1 ◽

2020 ◽

Author(s):

Jinghui Wang ◽

Zhu Yang ◽

Fengxi Long ◽

Li Luo ◽

Gao Li ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Cancer ◽

Human Liver ◽

Antitumor Effect ◽

Epithelial Mesenchymal Transition ◽

Epigenetic Silencing ◽

The Cancer Genome Atlas ◽

Mesenchymal Transition ◽

Human Liver Cancer

Abstract Background: The present study aimed to explore the role of ZIC4 in human liver cancer.Methods: Illumina450 genome-wide methylation data was downloaded from The Cancer Genome Atlas for 50 available liver tumor/surrounding pairs. Wound healing test, colony formation and flow cytometry assay were utilized to analyze cell migration, survival and apoptosis. The effects of EZH2 and ZIC4 on tumor growth were also investigated through in vivo xenograft and orthotopic implantation experiments. Results: ZIC4 was hypermethylated in liver cancer tissues and cell lines. EZH2 knockdown and DZNep mediated H3K27me3 contributes to ZIC4 expression. The antitumor effect of EZH2 knockdown on hepatocellular carcinoma growth, metastasis and epithelial-mesenchymal transition progression in vitro were rescued by sh-ZIC4. Downregulation of ZIC4 also rescued the antitumor effect of DZNep in vivo.Conclusions: Epigenetic silencing of ZIC4 by EZH2 mediated H3K27me3 is an important mechanism in human liver cancer and provide a new therapeutic target for the treatment of hepatocellular carcinoma disease.

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