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Jorge Filmus - One of the best experts on this subject based on the ideXlab platform.
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processing by convertases is required for Glypican 3 induced inhibition of hedgehog signaling
Journal of Biological Chemistry, 2015Co-Authors: Mariana I Capurro, Wen Shi, Tomomi Izumikawa, Hiroshi Kitagawa, Jorge FilmusAbstract:Abstract Glypican-3 (GPC3) is one of the six members of the mammalian Glypican family. We have previously reported that GPC3 inhibits Hedgehog (Hh) signaling by competing with Patched (Ptc) for Hh binding. We also showed that GPC3 binds with high affinity to Hh through its core protein, but that it does not interact with Ptc. Several members of the Glypican family, including GPC3, are subjected to an endoproteolytic cleavage by the furin-like convertase family of endoproteases. Surprisingly, however, we have found that a mutant GPC3 that cannot be processed by convertases is as potent as wild-type GPC3 in stimulating Wnt activity in HCC cell lines and 293T cells, and in promoting HCC growth. In this study we show that processing by convertases is essential for GPC3-induced inhibition of Hh signaling. Moreover, we show that a convertase-resistant GPC3 stimulates Hh signaling by increasing the binding of this growth factor to Ptc. Consistent with this, we show that the convertase-resistant mutant binds to both Hh and Ptc through its heparan sulfate (HS) chains. Unexpectedly, we found that the mutant core protein does not bind to Hh. We also report that the convertase-resistant mutant GPC3 carries HS chains with a significantly higher degree of sulfation than those of wild-type GPC3. We propose that the structural changes generated by the lack of cleavage determine a change in the sulfation of the HS chains, and that these hypersulfated chains mediate the interaction of the mutant GPC3 with Ptc.
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Glypican 3 binds to frizzled and plays a direct role in the stimulation of canonical wnt signaling
Journal of Cell Science, 2014Co-Authors: Mariana I Capurro, Wen Shi, Tonya Martin, Jorge FilmusAbstract:Glypican-3 (GPC3) is a proteoglycan that is bound to the cell surface. It is expressed by most hepatocellular carcinomas (HCCs) but not by normal hepatocytes. GPC3 stimulates HCC growth by promoting canonical Wnt signaling. Because Glypicans interact with Wnts, it has been proposed that these proteoglycans stimulate signaling by increasing the amount of Wnt at the cell membrane, thus facilitating the interaction of this growth factor with its signaling receptor, Frizzled. However, in this study, we demonstrate that GPC3 plays a more direct role in the stimulation of Wnt signaling. Specifically, we show that, in addition to interacting with Wnt, GPC3 and Frizzled interact directly through the glycosaminoglycan chains of GPC3, indicating that this Glypican stimulates the formation of signaling complexes between Wnt and Frizzled. Consistent with this, we show that the binding of Wnt at the cell membrane triggers the endocytosis of a complex that includes Wnt, Frizzled and GPC3. Additional support for our model is provided by the finding that Glypican-6 (GPC6) inhibits canonical Wnt signaling, despite the fact that it binds to Wnt at the cell membrane.
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Glypican 3 a marker and a therapeutic target in hepatocellular carcinoma
FEBS Journal, 2013Co-Authors: Jorge Filmus, Mariana CapurroAbstract:Glypican-3 (GPC3) is a member of the Glypican family. Glypicans are proteoglycans that are attached to the cell surface by a glycosyl-phosphatidylinositol anchor. They regulate the signaling activity of several growth factors, including Wnts. This regulation is based on the ability of Glypicans to stimulate or inhibit the interaction of these growth factors with their respective signaling receptors. It has been clearly established that whereas GPC3 is expressed by most hepatocellular carcinomas (HCCs), this Glypican is not detected in normal and cirrhotic liver, or in benign hepatic lesions. Consequently, immunostaining of liver biopsies for GPC3 is currently being used by clinical pathologists to confirm HCC diagnosis when the malignant nature of the lesion is difficult to establish. In addition to being a marker of HCC, GPC3 plays a role in the progression of the disease. GPC3 promotes the growth of HCC by stimulating canonical Wnt signaling. It has been proposed that this stimulation is based on the ability of GPC3 to increase the binding of Wnt to its signaling receptor, Frizzled. Two therapeutic approaches for HCC that target GPC3 are currently being tested in phase II clinical trials. One of them is based on the use of a humanized GPC3 monoclonal antibody that inhibits the in vivo growth of HCC xenografts by inducing antibody-dependent cellular cytotoxicity. The second approach employs a vaccine that consists of two GPC3-derived peptides that induce cytotoxic T lymphocytes against these peptides. Targeting of GPC3 might offer a new tool for the treatment of HCC.
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soluble Glypican 3 inhibits the growth of hepatocellular carcinoma in vitro and in vivo
International Journal of Cancer, 2010Co-Authors: Sandra Zittermann, Wen Shi, Mariana I Capurro, Jorge FilmusAbstract:The heterogeneity of the molecular pathology of HCC poses a formidable obstacle to the development of non-cytotoxic therapies. Several pro-tumorigenic signaling pathways can be aberrantly activated in HCC, including those triggered by Wnts. Glypican-3 (GPC3), a membrane-bound heparan sulfate proteoglycan that is overexpressed in most HCCs, promotes the growth of these tumors by stimulating Wnt signaling. Because GPC3 binds with high affinity to Wnts, and its growth-promoting activity requires attachment to the cell membrane, we have hypothesized that a mutated GPC3 lacking the GPI anchoring domain (sGPC3) will block Wnt signaling and inhibit the growth of Wnt-dependent tumors. In addition, because sGPC3 displays heparan sulfate chains, this secreted Glypican could also inhibit HCC growth by blocking the activity of other heparin-binding growth factors. To test this hypothesis, HCC cell lines were infected with an sGPC3-expressing lentivirus or virus control, and the effect of sGPC3 on the in vitro and in vivo growth was investigated. In addition, the signaling pathways targeted by sGPC3 were identified. We observed that sGPC3-expressing cells had lower proliferation rate. In addition, sGPC3 significantly inhibited the in vivo growth of the Huh6, HepG2 and Huh7 HCC cell lines. sGPC3 blocked Wnt signaling in Huh6- and Huh7-derived tumors and Erk1/2 and Akt phosphorylation in tumors generated by Huh7 and HepG2 cells, respectively. An anti-angiogenic effect in Huh7 and HepG2-derived tumors was also observed. We conclude that sGPC3 can inhibit HCC tumorigenicity by blocking the activity of several pro-tumorigenic growth factors.
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expression pattern of Glypican 3 gpc3 during human embryonic and fetal development
Histology and Histopathology, 2008Co-Authors: Bibiana V Iglesias, Jorge Filmus, Gloria Centeno, Hector Pascuccelli, Flavia Ward, Maria Giselle Peters, Lydia Puricelli, Elisa Bal De Kier JoffeAbstract:Glypicans represent a family of cell surface proteoglycans. Loss-of-function mutations in the human Glypican-3 (GPC3) gene results in the Simpson-Golabi-Behmel syndrome, characterized by severe malformations and pre- and postnatal overgrowth. Because the expression of GPC3 during human embryonic and fetal periods remains largely unknown, we investigated by immunohistochemistry its pattern of expression during four periods of human development covering the embryonic period (P1) from 5 to 8 weeks of development, and the fetal periods (P2, P3 and P4) from 9 to 28 weeks of development. Hepatocytes were homogeneously positive for GPC3 during the four periods while pancreatic acini and ducts showed a rather high staining only during P1. GPC3 was also detected in several kidney structures and in the genital system where the sex cords were weakly positive in P1 and P2. In later developmental stages the male's genital system expressed GPC3 while the female's did not. While the mesenchyme in the limbs showed positive staining in P1, GPC3 was not detected during the following stages. The mesenchymal tissue localized between the most caudal vertebrae was also positive in P1. A strong GPC3 signal was observed in neurons of the spinal cord and dorsal root ganglia in P2 and P3, while the brain was negative. In sum our studies revealed that GPC3 expression is highly tissue- and stage-specific during human development. The expression pattern of GPC3 is consistent with the abnormalities seen in the Simpson-Golabi-Behmel syndrome.
Ximing J Yang - One of the best experts on this subject based on the ideXlab platform.
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Glypican 3 overexpression in primary and metastatic wilms tumors
Virchows Archiv, 2015Co-Authors: Debra L Zynger, Chunyan Luan, Maria S Tretiakova, Nicole K Andeen, Laura S Finn, Masha Kocherginsky, Bin Tean Teh, Ximing J YangAbstract:Glypican 3 (GPC3), a heparan sulfate proteoglycan, plays a role in cell growth and differentiation. Mutations of the GPC3 gene are responsible for Simpson-Golabi-Behmel syndrome, which is characterized by anomalies of postnatal overgrowth and an increased risk of developing pediatric malignancies, mostly Wilms tumor and liver cancer. In order to understand the possible role of GPC3 in renal development and Wilms tumor formation, we analyzed messenger RNA (mRNA) and protein levels of GPC3 in sporadic Wilms tumors and compared it to normal kidneys and other common renal epithelial tumors. By using Affymetrix HGU133 oligonucleotide gene expression microarray data from 191 renal tumors and 12 normal kidneys, we found significant overexpression of GPC3 in Wilms tumors (p < 0.01), with 3.5-fold higher expression in comparison to normal kidneys and 6.5-fold higher than any type of renal tumors. The GPC3 gene product in Wilms tumor was further evaluated by immunohistochemistry and quantified by an automated image analysis. Cytoplasmic and membranous GPC3 immunoreactivity was present in 77 % of primary Wilms tumors (23/30), 93 % of metastatic Wilms tumors (13/14), 50 % of metanephric adenomas (4/8), 33 % of congenital mesoblastic nephromas (2/6), 100 % of nephrogenic rests (11/11), and 100 % of fetal kidneys (5/5). GPC3 staining was predominantly identified in blastemal and epithelial components of Wilms tumors, similar to that of fetal non-neoplastic kidney. All adult renal tumors (n = 60) and normal kidneys (n = 15) were GPC3 negative. These findings suggest the utility of GPC3 in differential diagnosis and follow-up of Wilms tumors. Our data also indicate that GPC3 is an oncofetal protein with a potential therapeutic value.
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Glypican 3 has a higher sensitivity than alpha fetoprotein for testicular and ovarian yolk sac tumour immunohistochemical investigation with analysis of histological growth patterns
Histopathology, 2010Co-Authors: Debra L Zynger, Pauline M Chou, Chunyan Luan, John Mccallum, Ximing J YangAbstract:Zynger D L, McCallum J C, Luan C, Chou P M & Yang X J (2010) Histopathology 56, 750–757 Glypican 3 has a higher sensitivity than alpha-fetoprotein for testicular and ovarian yolk sac tumour: immunohistochemical investigation with analysis of histological growth patterns Aims: Glypican 3 (GPC3) has been reported to be overexpressed in yolk sac tumour (YST), but the sensitivity has not been compared with α-fetoprotein (AFP). YST can form numerous growth patterns and the expression of GPC3 in these patterns has not been studied. The aim was to address these aspects. Methods and results: Sections from testicular or ovarian YST were subjected to immunohistochemistry using GPC3 (n = 39) and AFP (n = 24). Overall immunoreactivity for each case and specific histological patterns were semiquantitatively evaluated (0–3+) and intensity of reactivity was scored (0–3). All cases expressed GPC3 (1+, 5%; 2+, 8%; 3+, 87%) with strong intensity (2.9). The majority expressed AFP (58%) but immunoreactivity was often focal (0, 42%; 1+, 33%; 2+, 25%) and intensity was low (1.0). Using GPC3, >75% of the microcystic (n = 38), macrocystic (n = 26), solid (n = 21), glandular-alveolar (n = 8), endodermal sinus (n = 7), polyvesicular vitelline (n = 5), enteric (n = 4) and micropapillary (n = 2) growth patterns displayed 2+ or 3+ positivity. Conclusions: YST can display a variety of growth patterns that can be confused with other germ cell tumour components. GPC3 detects all growth patterns tested and has a higher sensitivity for detecting YST than AFP.
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Glypican 3 as a useful diagnostic marker that distinguishes hepatocellular carcinoma from benign hepatocellular mass lesions
Archives of Pathology & Laboratory Medicine, 2008Co-Authors: Hanlin L Wang, Florencia Anatelli, Qihui Jim Zhai, Brian P Adley, Shangtian Chuang, Ximing J YangAbstract:Abstract Context.—Histopathologic distinction between hepatocellular carcinoma (HCC) and benign hepatocellular mass lesions, particularly hepatocellular adenoma, can sometimes be challenging. The currently available ancillary tools are suboptimal in terms of sensitivity and specificity. Objective.—To further characterize the diagnostic value of Glypican-3 (GPC3), a cell surface proteoglycan that has recently been shown to be overexpressed in HCC, in the distinction between HCC and benign hepatocellular mass lesions. Design.—A total of 221 surgically resected liver specimens were subjected to immunohistochemical staining using a monoclonal antibody specific for GPC3. These included 111 HCCs, 48 hepatocellular adenomas, 30 focal nodular hyperplasias, and 32 large regenerative nodules in the background of cirrhosis. Results.—Cytoplasmic, membranous, and canalicular staining for GPC3 was detected in 84 (75.7%) of the 111 HCCs, among which, 61 (72.6%) of the 84 cases exhibited diffuse immunoreactivity. In cont...
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expression of Glypican 3 in ovarian and extragonadal germ cell tumors
American Journal of Clinical Pathology, 2008Co-Authors: Debra L Zynger, Michael J Everton, Nikolay D Dimov, Pauline M Chou, Ximing J YangAbstract:Germ cell tumors (GCTs), rare malignancies that occur in a wide range of locations and display variable histologic patterns, may pose diagnostic challenges. Glypican 3 (GPC3), a membrane-bound heparan sulfate proteoglycan, has been shown to be a novel diagnostic marker in testicular GCT. However, GPC3 expression in ovarian and extragonadal GCT has not been reported. We evaluated GPC3 immunoreactivity in GCTs from 63 patients (57 children and 6 adults), including 14 ovarian and 20 extragonadal primary GCTs and 8 metastases along with 21 primary testicular GCTs for comparison. All 33 yolk sac tumors (YSTs) and both choriocarcinomas were immunoreactive for GPC3. In contrast, a minority of immature (4/10) and mature (4/35) teratomas were positive. No positivity was seen in 6 embryonal carcinomas or 5 germinomas. GPC3 is differentially expressed in ovarian and extragonadal GCTs, with expression predominantly observed in YSTs and choriocarcinoma.
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value of Glypican 3 immunostaining in the diagnosis of hepatocellular carcinoma on needle biopsy
American Journal of Clinical Pathology, 2008Co-Authors: Florencia Anatelli, Ximing J Yang, Shangtian Chuang, Hanlin L WangAbstract:The diagnostic value of Glypican 3 (GPC3) immunostaining on needle biopsy specimens has not been well assessed. In this study, 120 liver needle biopsy specimens, including 46 from cirrhotic livers and 74 hepatocellular carcinomas (HCCs), were immunohistochemically examined for expression of GPC3. The results showed strong cytoplasmic and membranous staining in 36 HCCs (49%), among which 20 cases (56%) showed diffuse immunoreactivity. None of the 46 cirrhotic livers exhibited positive GPC3 immunostaining. The nonneoplastic liver tissues (cirrhotic or noncirrhotic) that were present in the majority of the HCC cases were also completely negative for GPC3 expression. These data demonstrate that GPC3 is a reliable immunohistochemical marker for the diagnosis of HCC on needle biopsy specimens when positive. However, the detection rate in our series seems lower than that reported in studies using resection specimens as the study materials. Our findings emphasize that GPC3 immunoreactivity can be focal and that negative staining should not be viewed as evidence to exclude the diagnosis of HCC in challenging needle biopsy specimens.
Yasuharu Nishimura - One of the best experts on this subject based on the ideXlab platform.
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Embryonic Stem Cell–Derived Dendritic Cells Expressing Glypican-3, a Recently Identified Oncofetal Antigen, Induce Protective Immunity against Highly Metastatic Mouse Melanoma, B16-F10
Cancer research, 2006Co-Authors: Yutaka Motomura, Satoru Senju, Daiki Fukuma, Tetsuya Nakatsura, Hidetake Matsuyoshi, Shinya Hirata, Mikio Monji, Hiroyuki Komori, Hideo Baba, Yasuharu NishimuraAbstract:We have recently established a method to generate dendritic cells from mouse embryonic stem cells. By introducing exogenous genes into embryonic stem cells and subsequently inducing differentiation to dendritic cells (ES-DC), we can now readily generate transfectant ES-DC expressing the transgenes. A previous study revealed that the transfer of genetically modified ES-DC expressing a model antigen, ovalbumin, protected the recipient mice from a challenge with an ovalbumin-expressing tumor. In the present study, we examined the capacity of ES-DC expressing mouse homologue of human Glypican-3, a recently identified oncofetal antigen expressed in human melanoma and hepatocellular carcinoma, to elicit protective immunity against Glypican-3-expressing mouse tumors. CTLs specific to multiple Glypican-3 epitopes were primed by the in vivo transfer of Glypican-3-transfectant ES-DC (ES-DC-GPC3). The transfer of ES-DC-GPC3 protected the recipient mice from subsequent challenge with B16-F10 melanoma, naturally expressing Glypican-3, and with Glypican-3-transfectant MCA205 sarcoma. The treatment with ES-DC-GPC3 was also highly effective against i.v. injected B16-F10. No harmful side effects, such as autoimmunity, were observed for these treatments. The depletion experiments and immunohistochemical analyses suggest that both CD8+ and CD4+ T cells contributed to the observed antitumor effect. In conclusion, the usefulness of Glypican-3 as a target antigen for antimelanoma immunotherapy was thus shown in the mouse model using the ES-DC system. Human dendritic cells expressing Glypican-3 would be a promising means for therapy of melanoma and hepatocellular carcinoma. (Cancer Res 2006; 66(4): 2414-22)
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embryonic stem cell derived dendritic cells expressing Glypican 3 a recently identified oncofetal antigen induce protective immunity against highly metastatic mouse melanoma b16 f10
Cancer Research, 2006Co-Authors: Yutaka Motomura, Satoru Senju, Daiki Fukuma, Tetsuya Nakatsura, Hidetake Matsuyoshi, Shinya Hirata, Mikio Monji, Hiroyuki Komori, Hideo Baba, Yasuharu NishimuraAbstract:We have recently established a method to generate dendritic cells from mouse embryonic stem cells. By introducing exogenous genes into embryonic stem cells and subsequently inducing differentiation to dendritic cells (ES-DC), we can now readily generate transfectant ES-DC expressing the transgenes. A previous study revealed that the transfer of genetically modified ES-DC expressing a model antigen, ovalbumin, protected the recipient mice from a challenge with an ovalbumin-expressing tumor. In the present study, we examined the capacity of ES-DC expressing mouse homologue of human Glypican-3, a recently identified oncofetal antigen expressed in human melanoma and hepatocellular carcinoma, to elicit protective immunity against Glypican-3-expressing mouse tumors. CTLs specific to multiple Glypican-3 epitopes were primed by the in vivo transfer of Glypican-3-transfectant ES-DC (ES-DC-GPC3). The transfer of ES-DC-GPC3 protected the recipient mice from subsequent challenge with B16-F10 melanoma, naturally expressing Glypican-3, and with Glypican-3-transfectant MCA205 sarcoma. The treatment with ES-DC-GPC3 was also highly effective against i.v. injected B16-F10. No harmful side effects, such as autoimmunity, were observed for these treatments. The depletion experiments and immunohistochemical analyses suggest that both CD8+ and CD4+ T cells contributed to the observed antitumor effect. In conclusion, the usefulness of Glypican-3 as a target antigen for antimelanoma immunotherapy was thus shown in the mouse model using the ES-DC system. Human dendritic cells expressing Glypican-3 would be a promising means for therapy of melanoma and hepatocellular carcinoma. (Cancer Res 2006; 66(4): 2414-22)
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embryonic stem cell derived dendritic cells expressing Glypican 3 a recently identified oncofetal antigen induce protective immunity against highly metastatic mouse melanoma b16 f10
Cancer Research, 2006Co-Authors: Yutaka Motomura, Satoru Senju, Daiki Fukuma, Tetsuya Nakatsura, Hidetake Matsuyoshi, Shinya Hirata, Mikio Monji, Hiroyuki Komori, Hideo Baba, Yasuharu NishimuraAbstract:We have recently established a method to generate dendritic cells from mouse embryonic stem cells. By introducing exogenous genes into embryonic stem cells and subsequently inducing differentiation to dendritic cells (ES-DC), we can now readily generate transfectant ES-DC expressing the transgenes. A previous study revealed that the transfer of genetically modified ES-DC expressing a model antigen, ovalbumin, protected the recipient mice from a challenge with an ovalbumin-expressing tumor. In the present study, we examined the capacity of ES-DC expressing mouse homologue of human Glypican-3, a recently identified oncofetal antigen expressed in human melanoma and hepatocellular carcinoma, to elicit protective immunity against Glypican-3-expressing mouse tumors. CTLs specific to multiple Glypican-3 epitopes were primed by the in vivo transfer of Glypican-3-transfectant ES-DC (ES-DC-GPC3). The transfer of ES-DC-GPC3 protected the recipient mice from subsequent challenge with B16-F10 melanoma, naturally expressing Glypican-3, and with Glypican-3-transfectant MCA205 sarcoma. The treatment with ES-DC-GPC3 was also highly effective against i.v. injected B16-F10. No harmful side effects, such as autoimmunity, were observed for these treatments. The depletion experiments and immunohistochemical analyses suggest that both CD8+ and CD4+ T cells contributed to the observed antitumor effect. In conclusion, the usefulness of Glypican-3 as a target antigen for antimelanoma immunotherapy was thus shown in the mouse model using the ES-DC system. Human dendritic cells expressing Glypican-3 would be a promising means for therapy of melanoma and hepatocellular carcinoma. (Cancer Res 2006; 66(4): 2414-22)
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usefulness of the novel oncofetal antigen Glypican 3 for diagnosis of hepatocellular carcinoma and melanoma
BioDrugs, 2005Co-Authors: Tetsuya Nakatsura, Yasuharu NishimuraAbstract:Glypican-3 (GPC3) mRNA and protein are expressed in >80% of human hepatocellular carcinomas (HCC) but not in normal tissues except for placenta and fetal liver. The oncofetal antigen GPC3 is a glycosylphosphatidyl inositol-anchored membrane protein and may be secreted. It is a novel tumor marker for human HCC: GPC3 protein was present in sera from 40–50% of HCC patients, but was not detected in sera from patients with liver cirrhosis or chronic hepatitis, or in sera from healthy individuals. α-Fetoprotein (AFP) and PIVKA-II (protein induced by vitamin K absence or antagonist-II), are well known major tumor markers for HCC. Generally, AFP shows high positivity for HCC but also high false-positivity in detection assays. Lens culinaris agglutinin-reactive fraction of α-fetoprotein (AFP-L3) is a recently described marker of HCC. Detection of AFP-L3 shows a much higher specificity than AFP, but a lower sensitivity. On the other hand, detection of PIVKA-II shows a lower false-positivity, but is not always sensitive enough to detect low levels secreted by small HCCs. There was no correlation between the three tumor markers, AFP, PIVKA-II, and GPC3 in terms of their presence in HCC cells. All three tumor markers showed similar positivity in patients with HCC, detecting 80% of patients with the disease.
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identification of Glypican 3 as a novel tumor marker for melanoma
Clinical Cancer Research, 2004Co-Authors: Tetsuya Nakatsura, Satoru Senju, Mikio Monji, Toshiro Kageshita, Shosuke Ito, Kazumasa Wakamatsu, Yoshiaki Ikuta, Tomomichi Ono, Yasuharu NishimuraAbstract:Purpose: We reported recently the novel tumor marker Glypican-3 (GPC3) for hepatocellular carcinoma. In the present study, we investigated the expression of GPC3 in human melanoma cell lines and tissues and asked whether GPC3 could be a novel tumor marker for melanoma. Experimental Design: Expression of GPC3 mRNA and protein was investigated in human melanoma cell lines and tissues using reverse transcription-PCR and immunohistochemical analysis. Secreted GPC3 protein was quantified using ELISA in culture supernatants of melanoma cell lines and in sera from 91 patients with melanoma and 28 diseasefree patients after surgical removal of primary melanoma. All of the subjects were Japanese nationals. Results: In >80% of melanoma and melanocytic nevus, there was evident expression of GPC3 mRNA and protein. Furthermore, GPC3 protein was evidenced in sera of 39.6% (36 of 91) of melanoma patients but not in sera from subjects with large congenital melanocytic nevus (0 of 5) and from healthy donors (0 of 60). Twenty-seven of 36 serum GPC3positive patients were negative for both serum 5-S-cysteinyldopa and melanoma-inhibitory activity, well-known tumor markers for melanoma. The positive rate of serum GPC3 (39.6%) was significantly higher than that of 5-S-cysteinyldopa (26.7%) and of melanoma-inhibitory activity (20.9%). Surprisingly, we detected serum GPC3 even in patients with stage 0 in situ melanoma. The positive rate of serum GPC3 at stage 0, I, and II (44.4%, 40.0%, and 47.6%) was significantly higher than that of 5-S-cysteinyldopa (0.0%, 8.0%, and 10.0%). Also observed was the disappearance of GPC3 protein in sera from 11 patients after surgical removal of the melanoma. Conclusions: GPC3 is apparently a novel tumor marker useful for the diagnosis of melanoma, especially in early stages of the disorder.
Tetsuya Nakatsura - One of the best experts on this subject based on the ideXlab platform.
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Next-Generation Cancer Immunotherapy Targeting Glypican-3
Frontiers Media S.A., 2019Co-Authors: Yasuhiro Shimizu, Toshiaki Yoshikawa, Toshihiro Suzuki, Itaru Endo, Tetsuya NakatsuraAbstract:Glypican-3 (GPC3), a 65 kD protein consisting of 580 amino acids, is a heparan sulfate proteoglycan bound to the cell membrane by glycosylphosphatidylinositol. This protein is expressed in the liver and the kidney of healthy fetuses but is hardly expressed in adults, except in the placenta. Contrarily, GPC3 is specifically expressed in hepatocellular carcinoma (HCC), ovarian clear cell carcinoma, melanoma, squamous cell carcinoma of the lung, hepatoblastoma, nephroblastoma (Wilms tumor), yolk sac tumor, and some pediatric cancers. Although the precise function of GPC3 remains unclear, it has been strongly suggested that it is related to the malignant transformation of HCC. We identified GPC3 as a promising target for cancer immunotherapy and have been working on the development of cancer immunotherapeutic agents targeting it through clinical trials. In some trials, it was revealed that the GPC3 peptide vaccines we developed using human leukocyte antigen-A24- and A2-restricted GPC3-derived peptides could induce GPC3-specific cytotoxic T cells in most vaccinated patients and thereby improve their prognosis. To further improve the clinical efficacy of cancer immunotherapy targeting GPC3, we are also developing next-generation therapeutic strategies using T cells engineered to express antigen-specific T-cell receptor or chimeric antigen receptor. In addition, we have successfully monitored the levels of serum full-length GPC3 protein, which is somehow secreted in the blood. The utility of GPC3 as a biomarker for predicting tumor recurrence and treatment efficacy is now being considered. In this review article, we summarize the results of clinical trials carried out by our team and describe the novel agent targeting the cancer-specific shared antigen, GPC3
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a Glypican 3 derived peptide vaccine against hepatocellular carcinoma
OncoImmunology, 2012Co-Authors: Yu Sawada, Mayuko Sakai, Toshiaki Yoshikawa, Kazuya Ofuji, Tetsuya NakatsuraAbstract:The results of a Phase I clinical trial in which a Glypican-3 (GPC3)-derived peptide was tested in advanced hepatocellular carcinoma patients point to a strong correlation between immunological and clinical responses. This commentary reviews our fundamental studies and clinical trials on the GPC3-derived peptide vaccine.
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embryonic stem cell derived dendritic cells expressing Glypican 3 a recently identified oncofetal antigen induce protective immunity against highly metastatic mouse melanoma b16 f10
Cancer Research, 2006Co-Authors: Yutaka Motomura, Satoru Senju, Daiki Fukuma, Tetsuya Nakatsura, Hidetake Matsuyoshi, Shinya Hirata, Mikio Monji, Hiroyuki Komori, Hideo Baba, Yasuharu NishimuraAbstract:We have recently established a method to generate dendritic cells from mouse embryonic stem cells. By introducing exogenous genes into embryonic stem cells and subsequently inducing differentiation to dendritic cells (ES-DC), we can now readily generate transfectant ES-DC expressing the transgenes. A previous study revealed that the transfer of genetically modified ES-DC expressing a model antigen, ovalbumin, protected the recipient mice from a challenge with an ovalbumin-expressing tumor. In the present study, we examined the capacity of ES-DC expressing mouse homologue of human Glypican-3, a recently identified oncofetal antigen expressed in human melanoma and hepatocellular carcinoma, to elicit protective immunity against Glypican-3-expressing mouse tumors. CTLs specific to multiple Glypican-3 epitopes were primed by the in vivo transfer of Glypican-3-transfectant ES-DC (ES-DC-GPC3). The transfer of ES-DC-GPC3 protected the recipient mice from subsequent challenge with B16-F10 melanoma, naturally expressing Glypican-3, and with Glypican-3-transfectant MCA205 sarcoma. The treatment with ES-DC-GPC3 was also highly effective against i.v. injected B16-F10. No harmful side effects, such as autoimmunity, were observed for these treatments. The depletion experiments and immunohistochemical analyses suggest that both CD8+ and CD4+ T cells contributed to the observed antitumor effect. In conclusion, the usefulness of Glypican-3 as a target antigen for antimelanoma immunotherapy was thus shown in the mouse model using the ES-DC system. Human dendritic cells expressing Glypican-3 would be a promising means for therapy of melanoma and hepatocellular carcinoma. (Cancer Res 2006; 66(4): 2414-22)
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Embryonic Stem Cell–Derived Dendritic Cells Expressing Glypican-3, a Recently Identified Oncofetal Antigen, Induce Protective Immunity against Highly Metastatic Mouse Melanoma, B16-F10
Cancer research, 2006Co-Authors: Yutaka Motomura, Satoru Senju, Daiki Fukuma, Tetsuya Nakatsura, Hidetake Matsuyoshi, Shinya Hirata, Mikio Monji, Hiroyuki Komori, Hideo Baba, Yasuharu NishimuraAbstract:We have recently established a method to generate dendritic cells from mouse embryonic stem cells. By introducing exogenous genes into embryonic stem cells and subsequently inducing differentiation to dendritic cells (ES-DC), we can now readily generate transfectant ES-DC expressing the transgenes. A previous study revealed that the transfer of genetically modified ES-DC expressing a model antigen, ovalbumin, protected the recipient mice from a challenge with an ovalbumin-expressing tumor. In the present study, we examined the capacity of ES-DC expressing mouse homologue of human Glypican-3, a recently identified oncofetal antigen expressed in human melanoma and hepatocellular carcinoma, to elicit protective immunity against Glypican-3-expressing mouse tumors. CTLs specific to multiple Glypican-3 epitopes were primed by the in vivo transfer of Glypican-3-transfectant ES-DC (ES-DC-GPC3). The transfer of ES-DC-GPC3 protected the recipient mice from subsequent challenge with B16-F10 melanoma, naturally expressing Glypican-3, and with Glypican-3-transfectant MCA205 sarcoma. The treatment with ES-DC-GPC3 was also highly effective against i.v. injected B16-F10. No harmful side effects, such as autoimmunity, were observed for these treatments. The depletion experiments and immunohistochemical analyses suggest that both CD8+ and CD4+ T cells contributed to the observed antitumor effect. In conclusion, the usefulness of Glypican-3 as a target antigen for antimelanoma immunotherapy was thus shown in the mouse model using the ES-DC system. Human dendritic cells expressing Glypican-3 would be a promising means for therapy of melanoma and hepatocellular carcinoma. (Cancer Res 2006; 66(4): 2414-22)
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embryonic stem cell derived dendritic cells expressing Glypican 3 a recently identified oncofetal antigen induce protective immunity against highly metastatic mouse melanoma b16 f10
Cancer Research, 2006Co-Authors: Yutaka Motomura, Satoru Senju, Daiki Fukuma, Tetsuya Nakatsura, Hidetake Matsuyoshi, Shinya Hirata, Mikio Monji, Hiroyuki Komori, Hideo Baba, Yasuharu NishimuraAbstract:We have recently established a method to generate dendritic cells from mouse embryonic stem cells. By introducing exogenous genes into embryonic stem cells and subsequently inducing differentiation to dendritic cells (ES-DC), we can now readily generate transfectant ES-DC expressing the transgenes. A previous study revealed that the transfer of genetically modified ES-DC expressing a model antigen, ovalbumin, protected the recipient mice from a challenge with an ovalbumin-expressing tumor. In the present study, we examined the capacity of ES-DC expressing mouse homologue of human Glypican-3, a recently identified oncofetal antigen expressed in human melanoma and hepatocellular carcinoma, to elicit protective immunity against Glypican-3-expressing mouse tumors. CTLs specific to multiple Glypican-3 epitopes were primed by the in vivo transfer of Glypican-3-transfectant ES-DC (ES-DC-GPC3). The transfer of ES-DC-GPC3 protected the recipient mice from subsequent challenge with B16-F10 melanoma, naturally expressing Glypican-3, and with Glypican-3-transfectant MCA205 sarcoma. The treatment with ES-DC-GPC3 was also highly effective against i.v. injected B16-F10. No harmful side effects, such as autoimmunity, were observed for these treatments. The depletion experiments and immunohistochemical analyses suggest that both CD8+ and CD4+ T cells contributed to the observed antitumor effect. In conclusion, the usefulness of Glypican-3 as a target antigen for antimelanoma immunotherapy was thus shown in the mouse model using the ES-DC system. Human dendritic cells expressing Glypican-3 would be a promising means for therapy of melanoma and hepatocellular carcinoma. (Cancer Res 2006; 66(4): 2414-22)
Debra L Zynger - One of the best experts on this subject based on the ideXlab platform.
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Glypican 3 overexpression in primary and metastatic wilms tumors
Virchows Archiv, 2015Co-Authors: Debra L Zynger, Chunyan Luan, Maria S Tretiakova, Nicole K Andeen, Laura S Finn, Masha Kocherginsky, Bin Tean Teh, Ximing J YangAbstract:Glypican 3 (GPC3), a heparan sulfate proteoglycan, plays a role in cell growth and differentiation. Mutations of the GPC3 gene are responsible for Simpson-Golabi-Behmel syndrome, which is characterized by anomalies of postnatal overgrowth and an increased risk of developing pediatric malignancies, mostly Wilms tumor and liver cancer. In order to understand the possible role of GPC3 in renal development and Wilms tumor formation, we analyzed messenger RNA (mRNA) and protein levels of GPC3 in sporadic Wilms tumors and compared it to normal kidneys and other common renal epithelial tumors. By using Affymetrix HGU133 oligonucleotide gene expression microarray data from 191 renal tumors and 12 normal kidneys, we found significant overexpression of GPC3 in Wilms tumors (p < 0.01), with 3.5-fold higher expression in comparison to normal kidneys and 6.5-fold higher than any type of renal tumors. The GPC3 gene product in Wilms tumor was further evaluated by immunohistochemistry and quantified by an automated image analysis. Cytoplasmic and membranous GPC3 immunoreactivity was present in 77 % of primary Wilms tumors (23/30), 93 % of metastatic Wilms tumors (13/14), 50 % of metanephric adenomas (4/8), 33 % of congenital mesoblastic nephromas (2/6), 100 % of nephrogenic rests (11/11), and 100 % of fetal kidneys (5/5). GPC3 staining was predominantly identified in blastemal and epithelial components of Wilms tumors, similar to that of fetal non-neoplastic kidney. All adult renal tumors (n = 60) and normal kidneys (n = 15) were GPC3 negative. These findings suggest the utility of GPC3 in differential diagnosis and follow-up of Wilms tumors. Our data also indicate that GPC3 is an oncofetal protein with a potential therapeutic value.
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Glypican 3 has a higher sensitivity than alpha fetoprotein for testicular and ovarian yolk sac tumour immunohistochemical investigation with analysis of histological growth patterns
Histopathology, 2010Co-Authors: Debra L Zynger, Pauline M Chou, Chunyan Luan, John Mccallum, Ximing J YangAbstract:Zynger D L, McCallum J C, Luan C, Chou P M & Yang X J (2010) Histopathology 56, 750–757 Glypican 3 has a higher sensitivity than alpha-fetoprotein for testicular and ovarian yolk sac tumour: immunohistochemical investigation with analysis of histological growth patterns Aims: Glypican 3 (GPC3) has been reported to be overexpressed in yolk sac tumour (YST), but the sensitivity has not been compared with α-fetoprotein (AFP). YST can form numerous growth patterns and the expression of GPC3 in these patterns has not been studied. The aim was to address these aspects. Methods and results: Sections from testicular or ovarian YST were subjected to immunohistochemistry using GPC3 (n = 39) and AFP (n = 24). Overall immunoreactivity for each case and specific histological patterns were semiquantitatively evaluated (0–3+) and intensity of reactivity was scored (0–3). All cases expressed GPC3 (1+, 5%; 2+, 8%; 3+, 87%) with strong intensity (2.9). The majority expressed AFP (58%) but immunoreactivity was often focal (0, 42%; 1+, 33%; 2+, 25%) and intensity was low (1.0). Using GPC3, >75% of the microcystic (n = 38), macrocystic (n = 26), solid (n = 21), glandular-alveolar (n = 8), endodermal sinus (n = 7), polyvesicular vitelline (n = 5), enteric (n = 4) and micropapillary (n = 2) growth patterns displayed 2+ or 3+ positivity. Conclusions: YST can display a variety of growth patterns that can be confused with other germ cell tumour components. GPC3 detects all growth patterns tested and has a higher sensitivity for detecting YST than AFP.
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expression of Glypican 3 in ovarian and extragonadal germ cell tumors
American Journal of Clinical Pathology, 2008Co-Authors: Debra L Zynger, Michael J Everton, Nikolay D Dimov, Pauline M Chou, Ximing J YangAbstract:Germ cell tumors (GCTs), rare malignancies that occur in a wide range of locations and display variable histologic patterns, may pose diagnostic challenges. Glypican 3 (GPC3), a membrane-bound heparan sulfate proteoglycan, has been shown to be a novel diagnostic marker in testicular GCT. However, GPC3 expression in ovarian and extragonadal GCT has not been reported. We evaluated GPC3 immunoreactivity in GCTs from 63 patients (57 children and 6 adults), including 14 ovarian and 20 extragonadal primary GCTs and 8 metastases along with 21 primary testicular GCTs for comparison. All 33 yolk sac tumors (YSTs) and both choriocarcinomas were immunoreactive for GPC3. In contrast, a minority of immature (4/10) and mature (4/35) teratomas were positive. No positivity was seen in 6 embryonal carcinomas or 5 germinomas. GPC3 is differentially expressed in ovarian and extragonadal GCTs, with expression predominantly observed in YSTs and choriocarcinoma.
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expression of Glypican 3 in hepatoblastoma an immunohistochemical study of 65 cases
Human Pathology, 2008Co-Authors: Debra L Zynger, Pauline M Chou, Ximing J Yang, Anita Gupta, Chunyan Luan, Guang Yu YangAbstract:Glypican 3 is a heparin sulfate proteoglycan bound to the cell surface that is theorized to participate in cell signaling, resulting in embryonic cell growth and differentiation. The GPC3 gene is mutated in Simpson-Golabi-Behmel syndrome, whose features include numerous developmental abnormalities, tissue overgrowth, and an increased risk for embryonal malignancies, including hepatoblastoma. GPC3 has been detected in hepatic stem cells and was recently identified as one of the most overexpressed genes in hepatoblastoma by microarray analysis. The purpose of this study was to analyze the expression of GPC3 in a large series of hepatoblastoma using immunohistochemistry to assess its use as a diagnostic marker. The GPC3 immunoreactivity was semiquantitatively evaluated in 65 cases of hepatoblastoma. In addition, histologic patterns in each tumor were individually assessed for immunoreactivity. All 65 hepatoblastomas had cytoplasmic immunoreactivity for GPC3 with greater than 90% of cases showing strong, diffuse positivity. There was no reactivity in benign liver tissue. Fetal, embryonal, and small cell undifferentiated patterns were diffusely positive in almost all cases, whereas mesenchymal and teratoid patterns were nearly all negative. The high GPC3 expression consistently demonstrated in this study suggests that GPC3 may play a role in the tumorigenesis of hepatoblastoma.
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Glypican 3 a novel marker in testicular germ cell tumors
The American Journal of Surgical Pathology, 2006Co-Authors: Debra L Zynger, Nikolay D Dimov, Chunyan Luan, Bin Tean Teh, Ximing J YangAbstract:Glypican 3 (GPC3), a membrane-bound heparin sulfate proteoglycan, may play a role in promoting embryonic cell growth and differentiation. GPC3 is mutated in Simpson-Golabi-Behmel syndrome, characterized by tissue overgrowth and an increased risk of embryonal malignancies. Recently, GPC3 was reported to be one of the over-expressed genes in testicular yolk sac tumors by gene expression microarray analysis. However, the presence of the GPC3 protein in germ cell tumors has never been investigated. The purpose of the study was to investigate the GPC3 expression in various histologic components of testicular germ cell tumors using immunohistochemistry and to assess its possible utility as a diagnostic marker. Tumors from 71 patients were examined, including components of 42 seminomas, 37 embryonal carcinomas, 24 yolk sac tumors, 20 teratomas with mature elements, 16 teratomas with immature elements, and 7 choriocarcinomas. Cytoplasmic and membranous immunoreactivity was semiquantitatively evaluated. All yolk sac tumor (24/24) and choriocarcinoma (7/7) components were immunoreactive for GPC3, whereas only 38% of teratomas with immature elements and 8% of embryonal carcinomas expressed GPC3. There was no immunoreactivity in benign testicular tissue, intratubular germ cell neoplasia, seminomas (0/42), or teratomas with mature elements (0/20). We conclude that the oncofetal protein GPC3 is a novel immunohistochemical marker in testicular germ cell tumors with differential expression in defined histologic subtypes. Our findings suggest a possible role of GPC3 in tumor cell differentiation. Furthermore, GPC immunostaining may be useful in the pathologic diagnosis of nonseminomatous germ cell tumors, particularly yolk sac tumor, and choriocarcinoma.