Glypican 3 Explained

Glypican-3 is a protein that, in humans, is encoded by the GPC3 gene.[1] [2] [3] [4] The GPC3 gene is located on human X chromosome (Xq26) where the most common gene (Isoform 2, GenBank Accession No.: NP_004475) encodes a 70-kDa core protein with 580 amino acids.[5] Three variants have been detected that encode alternatively spliced forms termed Isoforms 1 (NP_001158089), Isoform 3 (NP_001158090) and Isoform 4 (NP_001158091).

Structure and function

The protein core of GPC3 consists of two subunits, where the N-terminal subunit has a size of ~40 kDa and the C-terminal subunit is ~30 kDa. Six glypicans (GPC1-6) have been identified in mammals. Cell surface heparan sulfate proteoglycans are composed of a membrane-associated protein core substituted with a variable number of heparan sulfate chains. Members of the glypican-related integral membrane proteoglycan family (GRIPS) contain a core protein anchored to the cytoplasmic membrane via a glycosyl phosphatidylinositol linkage. These proteins may play a role in the control of cell division and growth regulation.[3] GPC3 has been found to regulate Wnt/β-catenin and Yap signaling pathways.[6] GPC3 interacts with both Wnt and frizzled (FZD) to form a complex and triggers downstream signaling.[7] [8] The core protein of GPC3 may serve as a co-receptor or a receiver for Wnt. A cysteine-rich domain at the N-lobe of GPC3 has been identified as a hydrophobic groove that interacts with Wnt3a. Blocking the Wnt binding domain on GPC3 using the HN3 single domain antibody can inhibit Wnt activation. Wnt also recognizes a heparan sulfate structure on GPC3, which contains IdoA2S and GlcNS6S, and that the 3-O-sulfation in GlcNS6S3S significantly enhances the binding of Wnt to heparan sulfate.[9] GPC3 also modulates Yap signaling. It might interact with FAT1 on the cell surface.[10]

Disease linkage

Deletion mutations in this gene are associated with Simpson–Golabi–Behmel syndrome.[1]

Diagnostic utility

Glypican 3 immunostaining has utility for differentiating hepatocellular carcinoma (HCC)[11] and dysplastic changes in cirrhotic livers; HCC stains with glypican 3, while liver with dysplastic changes and/or cirrhotic changes does not.[12] Using the YP7 murine monoclonal antibody, GPC3 protein expression is found in HCC, not in normal liver and cholangiocarcinoma.[13] The YP7 murine antibody has been humanized and named as 'hYP7'.[14] GPC3 is also expressed to a lesser degree in melanoma, ovarian clear-cell carcinomas, yolk sac tumors, neuroblastoma, hepatoblastoma, Wilms' tumor cells, and other tumors. However, the significance of GPC3 as a diagnostic tool for human tumors other than HCC is unclear.

Therapeutic potential

To validate GPC3 as a therapeutic target in liver cancer, the anti-GPC3 therapeutic antibodies GC33,[15] YP7, HN3 and HS20[16] have been made and widely tested. The laboratory of Mitchell Ho at the National Cancer Institute, NIH (Bethesda, Maryland, US) has generated YP7 and other murine monoclonal antibodies that recognize the C-lobe of GPC3 by hybridoma technology. These antibodies have been humanized (e.g. hYP7) via antibody engineering for clinical applications. The Ho lab has also identified the human single-domain antibody ('human nanobody') HN3[17] targeting the N-lobe of GPC3 and the human monoclonal antibody HS20[18] [19] targeting the heparan sulfate chains on GPC3 by phage display technology. Both HN3 and HS20 antibodies inhibit Wnt signaling in liver cancer cells . The immunotoxins based on HN3,[20] [21] [22] the antibody-drug conjugates based on hYP7[23] and the T-cell engaging bispecific antibodies derived from YP7[24] [25] and GC33,[26] have been developed for treating liver cancer. The chimeric antigen receptor (CAR) T cell immunotherapies based on GC33,[27] hYP7[28] [29] and HN3[30] are being developed at various stages for treating liver cancer. In mice with xenograft or orthoptic liver tumors, CAR (hYP7) T cells can eliminate GPC3-positive cancer cells, by inducing perforin- and granzyme-mediated cell death and reducing Wnt signaling in tumor cells. CAR (hYP7) T cells are being evaluated at a clinical trial at the NIH.[31]

See also

Further reading

External links

Notes and References

  1. Pilia G, Hughes-Benzie RM, MacKenzie A, Baybayan P, Chen EY, Huber R, Neri G, Cao A, Forabosco A, Schlessinger D . 6 . Mutations in GPC3, a glypican gene, cause the Simpson-Golabi-Behmel overgrowth syndrome . Nature Genetics . 12 . 3 . 241–247 . March 1996 . 8589713 . 10.1038/ng0396-241 . 38846721 .
  2. Veugelers M, Vermeesch J, Watanabe K, Yamaguchi Y, Marynen P, David G . GPC4, the gene for human K-glypican, flanks GPC3 on xq26: deletion of the GPC3-GPC4 gene cluster in one family with Simpson-Golabi-Behmel syndrome . Genomics . 53 . 1 . 1–11 . October 1998 . 9787072 . 10.1006/geno.1998.5465 .
  3. Web site: Entrez Gene: GPC3 glypican 3.
  4. Jakubovic BD, Jothy S . Glypican-3: from the mutations of Simpson-Golabi-Behmel genetic syndrome to a tumor marker for hepatocellular carcinoma . Experimental and Molecular Pathology . 82 . 2 . 184–189 . April 2007 . 17258707 . 10.1016/j.yexmp.2006.10.010 .
  5. Ho M, Kim H . Glypican-3: a new target for cancer immunotherapy . European Journal of Cancer . 47 . 3 . 333–338 . February 2011 . 21112773 . 3031711 . 10.1016/j.ejca.2010.10.024 .
  6. Kolluri A, Ho M . The Role of Glypican-3 in Regulating Wnt, YAP, and Hedgehog in Liver Cancer . Frontiers in Oncology . 9 . 708 . 2019 . 31428581 . 6688162 . 10.3389/fonc.2019.00708 . free .
  7. Li N, Gao W, Zhang YF, Ho M . Glypicans as Cancer Therapeutic Targets . Trends in Cancer . 4 . 11 . 741–754 . November 2018 . 30352677 . 6209326 . 10.1016/j.trecan.2018.09.004 .
  8. Li N, Wei L, Liu X, Bai H, Ye Y, Li D, Li N, Baxa U, Wang Q, Lv L, Chen Y, Feng M, Lee B, Gao W, Ho M . 6 . A Frizzled-Like Cysteine-Rich Domain in Glypican-3 Mediates Wnt Binding and Regulates Hepatocellular Carcinoma Tumor Growth in Mice . Hepatology . 70 . 4 . 1231–1245 . October 2019 . 30963603 . 6783318 . 10.1002/hep.30646 .
  9. Gao W, Xu Y, Liu J, Ho M . Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate . Scientific Reports . 6 . 26245 . May 2016 . 27185050 . 4869111 . 10.1038/srep26245 . 2016NatSR...626245G .
  10. Meng P, Zhang YF, Zhang W, Chen X, Xu T, Hu S, Liang X, Feng M, Yang X, Ho M . 6 . Identification of the atypical cadherin FAT1 as a novel glypican-3 interacting protein in liver cancer cells . Scientific Reports . 11 . 1 . 40 . January 2021 . 33420124 . 7794441 . 10.1038/s41598-020-79524-3 .
  11. Filmus J, Capurro M . Glypican-3 and alphafetoprotein as diagnostic tests for hepatocellular carcinoma . Molecular Diagnosis . 8 . 4 . 207–212 . 2004 . 15887976 . 10.1007/bf03260065 . 6312940 .
  12. Anatelli F, Chuang ST, Yang XJ, Wang HL . Value of glypican 3 immunostaining in the diagnosis of hepatocellular carcinoma on needle biopsy . American Journal of Clinical Pathology . 130 . 2 . 219–223 . August 2008 . 18628090 . 10.1309/WMB5PX57Y4P8QCTY . 45888415 .
  13. Phung Y, Gao W, Man YG, Nagata S, Ho M . High-affinity monoclonal antibodies to cell surface tumor antigen glypican-3 generated through a combination of peptide immunization and flow cytometry screening . mAbs . 4 . 5 . 592–599 . September 2012 . 22820551 . 3499300 . 10.4161/mabs.20933 .
  14. Zhang YF, Ho M . Humanization of high-affinity antibodies targeting glypican-3 in hepatocellular carcinoma . Scientific Reports . 6 . 33878 . September 2016 . 27667400 . 5036187 . 10.1038/srep33878 . 2016NatSR...633878Z .
  15. 6 . Ishiguro T, Sugimoto M, Kinoshita Y, Miyazaki Y, Nakano K, Tsunoda H, Sugo I, Ohizumi I, Aburatani H, Hamakubo T, Kodama T, Tsuchiya M, Yamada-Okabe H . December 2008 . Anti-glypican 3 antibody as a potential antitumor agent for human liver cancer . Cancer Research . 68 . 23 . 9832–9838 . 10.1158/0008-5472.CAN-08-1973 . 19047163.
  16. Kim H, Ho M . Isolation of Antibodies to Heparan Sulfate on Glypicans by Phage Display . Current Protocols in Protein Science . 94 . 1 . e66 . November 2018 . 30091851 . 6205898 . 10.1002/cpps.66 .
  17. Feng M, Gao W, Wang R, Chen W, Man YG, Figg WD, Wang XW, Dimitrov DS, Ho M . 6 . Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma . Proceedings of the National Academy of Sciences of the United States of America . 110 . 12 . E1083–E1091 . March 2013 . 23471984 . 3607002 . 10.1073/pnas.1217868110 . free . 2013PNAS..110E1083F .
  18. Gao W, Kim H, Feng M, Phung Y, Xavier CP, Rubin JS, Ho M . Inactivation of Wnt signaling by a human antibody that recognizes the heparan sulfate chains of glypican-3 for liver cancer therapy . Hepatology . 60 . 2 . 576–587 . August 2014 . 24492943 . 4083010 . 10.1002/hep.26996 .
  19. Kim H, Ho M . Isolation of Antibodies to Heparan Sulfate on Glypicans by Phage Display . Current Protocols in Protein Science . 94 . 1 . e66 . November 2018 . 30091851 . 6205898 . 10.1002/cpps.66 .
  20. Gao W, Tang Z, Zhang YF, Feng M, Qian M, Dimitrov DS, Ho M . Immunotoxin targeting glypican-3 regresses liver cancer via dual inhibition of Wnt signalling and protein synthesis . Nature Communications . 6 . 6536 . March 2015 . 25758784 . 4357278 . 10.1038/ncomms7536 . 2015NatCo...6.6536G .
  21. Wang C, Gao W, Feng M, Pastan I, Ho M . Construction of an immunotoxin, HN3-mPE24, targeting glypican-3 for liver cancer therapy . Oncotarget . 8 . 20 . 32450–32460 . May 2017 . 27419635 . 5464801 . 10.18632/oncotarget.10592 .
  22. Fleming BD, Urban DJ, Hall MD, Longerich T, Greten TF, Pastan I, Ho M . Engineered Anti-GPC3 Immunotoxin, HN3-ABD-T20, Produces Regression in Mouse Liver Cancer Xenografts Through Prolonged Serum Retention . Hepatology . 71 . 5 . 1696–1711 . May 2020 . 31520528 . 7069773 . 10.1002/hep.30949 .
  23. Fu Y, Urban DJ, Nani RR, Zhang YF, Li N, Fu H, Shah H, Gorka AP, Guha R, Chen L, Hall MD, Schnermann MJ, Ho M . 6 . Glypican-3-Specific Antibody Drug Conjugates Targeting Hepatocellular Carcinoma . Hepatology . 70 . 2 . 563–576 . August 2019 . 30353932 . 6482108 . 10.1002/hep.30326 .
  24. Web site: Federal Register /Vol. 82, No. 96 / Friday, May 19, 2017.
  25. Chen X, Chen Y, Liang R, Xiang L, Li J, Zhu Y, He H, Huang L, Zuo D, Li W, Liang X, Dong S, Hu S, Ho M, Feng M . 6 . Combination Therapy of Hepatocellular Carcinoma by GPC3-Targeted Bispecific Antibody and Irinotecan is Potent in Suppressing Tumor Growth in Mice . Molecular Cancer Therapeutics . 149–158 . November 2021 . 21 . 1 . 34725191 . 10.1158/1535-7163.MCT-20-1025 . 8742776 .
  26. Ishiguro T, Sano Y, Komatsu SI, Kamata-Sakurai M, Kaneko A, Kinoshita Y, Shiraiwa H, Azuma Y, Tsunenari T, Kayukawa Y, Sonobe Y, Ono N, Sakata K, Fujii T, Miyazaki Y, Noguchi M, Endo M, Harada A, Frings W, Fujii E, Nanba E, Narita A, Sakamoto A, Wakabayashi T, Konishi H, Segawa H, Igawa T, Tsushima T, Mutoh H, Nishito Y, Takahashi M, Stewart L, ElGabry E, Kawabe Y, Ishigai M, Chiba S, Aoki M, Hattori K, Nezu J . 6 . An anti-glypican 3/CD3 bispecific T cell-redirecting antibody for treatment of solid tumors . Science Translational Medicine . 9 . 410 . eaal4291 . October 2017 . 28978751 . 10.1126/scitranslmed.aal4291 . 206693656 . free .
  27. Gao H, Li K, Tu H, Pan X, Jiang H, Shi B, Kong J, Wang H, Yang S, Gu J, Li Z . 6 . Development of T cells redirected to glypican-3 for the treatment of hepatocellular carcinoma . Clinical Cancer Research . 20 . 24 . 6418–6428 . December 2014 . 25320357 . 10.1158/1078-0432.CCR-14-1170 . 24474000 . free .
  28. Li D, Li N, Zhang Y, Fu H, Torres MB, Wang Q, Greten TF, Ho M . 2018-07-01. Abstract 2549: Development of CAR T-cell therapy targeting glypican-3 in liver cancer . Immunology. 78 . 13_Supplement . American Association for Cancer Research . 2549 . 10.1158/1538-7445.AM2018-2549 . 81043794.
  29. Li D, Li N, Zhang YF, Fu H, Feng M, Schneider D, Su L, Wu X, Zhou J, Mackay S, Kramer J, Duan Z, Yang H, Kolluri A, Hummer AM, Torres MB, Zhu H, Hall MD, Luo X, Chen J, Wang Q, Abate-Daga D, Dropulic B, Hewitt SM, Orentas RJ, Greten TF, Ho M . 6 . Persistent Polyfunctional Chimeric Antigen Receptor T Cells That Target Glypican 3 Eliminate Orthotopic Hepatocellular Carcinomas in Mice . Gastroenterology . 158 . 8 . 2250–2265.e20 . June 2020 . 32060001 . 7282931 . 10.1053/j.gastro.2020.02.011 .
  30. Kolluri . Aarti . Li . Dan . Li . Nan . Duan . Zhijian . Roberts . Lewis R. . Ho . Mitchell . 2023-02-01 . Human VH-based chimeric antigen receptor T cells targeting glypican 3 eliminate tumors in preclinical models of HCC . Hepatology Communications . 7 . 2 . e0022 . 10.1097/HC9.0000000000000022 . 2471-254X . 36691969. 9851680 . free .
  31. https://clinicaltrials.gov/ct2/show/NCT05003895 NCT05003895