Abstract
The glypican-3 (GPC3) receptor is overexpressed in hepatocellular carcinoma (HCC) and is a potential diagnostic and therapeutic target. GPC3-targeted molecular imaging will be helpful to differentiate diagnosis and guide therapy. In the present study, we will develop a novel PET probe for imaging the expression of GPC-3. L5 (sequence: RLNVGGTYFLTTRQ), a GPC3 targeting peptide, was labeled with 5-carboxyfluorescein (FAM) and 18F-fluoride. Cell binding tests were performed to identify the binding specificity of FAM-L5 and 18F radiolabeled peptide. MicroPET/CT imaging was used to determine the potential of a novel PET tracer for visualizing HCC tumors with a high expression of GPC3. In vitro binding tests showed that the uptake of FAM-L5 in HepG2 cells (high expression of GPC3) was significantly higher than that of HL-7702 cells (negative expression of GPC3) (mean fluorescent intensity: 14,094 ± 797 vs. 2765 ± 314 events, t = 32.363, P = 0.000). Confocal fluorescent imaging identified that FAM-L5 accumulated where the GPC3 receptor was located. A novel PET tracer (18F-AlF-NODA-MP-6-Aoc-L5) was successfully labeled by chelation chemistry. In vitro cell uptake studies showed that 18F-AlF-NODA-MP-6-Aoc-L5 can bind to HepG2 tumor cells and was stable in PBS and mouse serum stability tests. MicroPET/CT showed that HepG2 tumors could be clearly visualized with a tumor/muscle ratio of 2.46 ± 0.53. However, the tumor/liver ratio was low (0.93 ± 0.16) due to the high physiological uptake in the liver. This study demonstrates that FAM and the 18F-labeled L5 peptide can selectively target HCC with a high expression of GPC3 in vitro and in vivo. 18F-AlF-NODA-MP-C6-L5 has the potential to be a GPC3 target tracer but requires some chemical modifications to achieve a high enough tumor/liver ratio for detection of the tumor in the liver.
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Abbreviations
- GPC3:
-
Glypican-3
- HCC:
-
Hepatocellular carcinoma
- FAM:
-
5-Carboxyfluorescein
- TACE:
-
Transarterial radioembolization
- PET:
-
Positron emission tomography
- HPLC:
-
High-performance liquid chromatography
- DAPI:
-
4,6-Diamidino-2-phenylindole
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- PBS:
-
Phosphate-buffered saline
- OSEM:
-
Ordered subsets expectation maximum
- IRW:
-
Inveon Research Workplace
- ROI:
-
Region of interest
- VOI:
-
Volume of interest
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Acknowledgements
We wish to thank our colleagues at the Nanfang PET center and the Department of Oncology and Pathology at the Nanfang Hospital for their support. This work was supported by funding from the National Natural Science Foundation Project of China (81071175, 81371591) and the Natural Science Foundation Project of Guangdong Province (2014A030313311).
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National Natural Science Foundation Project of China (81071175, 81371591); Natural Science Foundation Project of Guangdong Province (2014A030313311).
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Wang, Z., Han, YJ., Huang, S. et al. Imaging the expression of glypican-3 in hepatocellular carcinoma by PET. Amino Acids 50, 309–320 (2018). https://doi.org/10.1007/s00726-017-2517-z
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DOI: https://doi.org/10.1007/s00726-017-2517-z