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Protein PEGylation for cancer therapy: bench to bedside

  • Vijayalaxmi Gupta
  • Sneha Bhavanasi
  • Mohiuddin Quadir
  • Kevin Singh
  • Gaurav Ghosh
  • Kritin Vasamreddy
  • Arnab Ghosh
  • Teruna J. Siahaan
  • Snigdha Banerjee
  • Sushanta K. Banerjee
Review
  • 106 Downloads

Abstract

PEGylation is a biochemical modification process of bioactive molecules with polyethylene glycol (PEG), which lends several desirable properties to proteins/peptides, antibodies, and vesicles considered to be used for therapy or genetic modification of cells. However, PEGylation of proteins is a complex process and can be carried out using more than one strategy that depends on the nature of the protein and the desired application. Proteins of interest are covalently conjugated or non-covalently complexed with inert PEG strings. Purification of PEGylated protein is another critical step, which is mainly carried out based on electrostatic interactions or molecular sizes using chromatography. Several PEGylated drugs are being used for diseases like anemia, kidney disease, multiple sclerosis, hemophilia and cancers. With the advancement and increased specificity of the PEGylation process, the world of drug therapy, and specifically cancer therapy could benefit by utilizing this technique to create more stable and non-immunogenic therapies. In this article we describe the structure and functions of PEGylation and how this chemistry helps in drug discovery. Moreover, special emphasis has been given to CCN-family proteins that can be targeted or used as therapy to prevent or block cancer progression through PEGylation technology.

Keywords

PEGylation Cancer Polyethylene glycol Nanoparticles Immunogenicity 

Abbreviations

PEG

Polyethylene glycol

NP

Nanoparticle

MW

Molecular weight

Notes

Acknowledgements

We thank the members of Kansas City VA Research Office and Midwest Biomedical Research Foundation Administrative and clerical supports.

Author’s contribution

Conception and design: S. K. Banerjee, S. Banerjee, M. Quadir, and V. Gupta, A. Ghosh; Writing and review of the manuscript: V. Gupta, M. Quadir, S. Bhavanasi, G. Ghosh, T. Siahaan, and S.K. Banerjee; Administrative, technical or material support: M. Quadir and S. K. Banerjee, and Study supervision: S. K. Banerjee.

Funding

The work is supported by Merit review grant from Department of Veterans Affairs (Sushanta K. Banerjee, 5I01BX001989–04 and Snigdha Banerjee, I01BX001002–05), KUMC Lied Basic Science Grant Program (SKB), and Grace Hortense Greenley Trust, directed by The Research Foundation in memory of Eva Lee Caldwell (SKB). This work is partially supported by NIH grant P20 GM109024 from the National Institute of General Medical Science (NIGMS) (MQ), NSF under Grant No.  092354 (MQ), NIH Grant Number 2P20 RR015566 from the National Center for Research Resources (MQ), NIH grant 1R01 GM 114080 (NIGMS) and NSF Grant No. IIA-1355466 from North Dakota Established Program to Stimulate Comperative Research (EPSCoR) through the Center for Sustanable Materials Science (MQ).

Compliance with ethical standards

Ethics approval and consent to participate

Compliance with ethical standard of VA Medical Center.

Consent for publication

All the authors of this manuscript have agreed to publish this article.

Competing interests

No potential conflicts of interest were disclosed.

References

  1. Abuchowski A, Kazo GM, Verhoest CR Jr et al (1984) Cancer therapy with chemically modified enzymes. I. Antitumor properties of polyethylene glycol-asparaginase conjugates. Cancer Biochem Biophys 7:175–186PubMedGoogle Scholar
  2. Bailon P, Won CY (2009) PEG-modified biopharmaceuticals. Expert Opin Drug Deliv 6:1–16CrossRefGoogle Scholar
  3. Banerjee S, Dhar G, Haque I et al (2008) CCN5/WISP-2 expression in breast adenocarcinoma is associated with less frequent progression of the disease and suppresses the invasive phenotypes of tumor cells. Cancer Res 68:7606–7612CrossRefGoogle Scholar
  4. Banerjee SK, Maity G, Haque I et al (2016) Human pancreatic cancer progression: an anarchy among CCN-siblings. Journal of cell communication and signaling 10:207–216CrossRefGoogle Scholar
  5. Bayes M, Rabasseda X, Prous JR (2002) Gateways to clinical trials. Methods Find Exp Clin Pharmacol 24:703–729PubMedGoogle Scholar
  6. Booth C, Gaspar HB (2009) Pegademase bovine (PEG-ADA) for the treatment of infants and children with severe combined immunodeficiency (SCID). Biologics 3:349–358PubMedPubMedCentralGoogle Scholar
  7. Carter PJ (2011) Introduction to current and future protein therapeutics: a protein engineering perspective. Exp Cell Res 317:1261–1269CrossRefGoogle Scholar
  8. Chen X, Hu C, Zhang W et al (2015) Metformin inhibits the proliferation, metastasis, and cancer stem-like sphere formation in osteosarcoma MG63 cells in vitro. Tumour biology: the journal of the International Society for Oncodevelopmental Biology and Medicine 36:9873–9883CrossRefGoogle Scholar
  9. Choy EH, Hazleman B, Smith M et al (2002) Efficacy of a novel PEGylated humanized anti-TNF fragment (CDP870) in patients with rheumatoid arthritis: a phase II double-blinded, randomized, dose-escalating trial. Rheumatology 41:1133–1137CrossRefGoogle Scholar
  10. Codelli JA, Baskin JM, Agard NJ et al (2008) Second-generation difluorinated cyclooctynes for copper-free click chemistry. J Am Chem Soc 130:11486–11493CrossRefGoogle Scholar
  11. Deiters A, Cropp T, Summerer D et al (2004) Site-specific PEGylation of proteins containing unnatural amino acids. Bioorg Med Chem Lett 14:5743–5745CrossRefGoogle Scholar
  12. Dhar G, Banerjee S, Dhar K et al (2008) Gain of oncogenic function of p53 mutants induces invasive phenotypes in human breast cancer cells by silencing CCN5/WISP-2. Cancer Res 68:4580–4587CrossRefGoogle Scholar
  13. Dhiman S, Mishra N, Sharma S (2016) Development of PEGylated solid lipid nanoparticles of pentoxifylline for their beneficial pharmacological potential in pathological cardiac hypertrophy. Artif Cells Nanomed Biotechnol 44:1901–1908CrossRefGoogle Scholar
  14. Dozier JK, Distefano MD (2015) Site-specific PEGylation of therapeutic proteins. Int J Mol Sci 16:25831–25864CrossRefGoogle Scholar
  15. Fernandes C, Benfeito S, Amorim R et al (2018) Desrisking the cytotoxicity of a Mitochondriotropic antioxidant based on Caffeic acid by a PEGylated strategy. Bioconjug Chem 29:2723–2733CrossRefGoogle Scholar
  16. Grayson SM, Godbey WT (2008) The role of macromolecular architecture in passively targeted polymeric carriers for drug and gene delivery. J Drug Target 16:329–356CrossRefGoogle Scholar
  17. Haque I, Banerjee S, Mehta S et al (2011a) Cysteine-rich 61-connective tissue growth factor-nephroblastoma-overexpressed 5 (CCN5)/Wnt-1-induced signaling protein-2 (WISP-2) regulates microRNA-10b via hypoxia-inducible factor-1alpha-TWIST signaling networks in human breast cancer cells. JBiolChem 286:43475–43485Google Scholar
  18. Haque I, Mehta S, Majumder M et al (2011b) Cyr61/CCN1 signaling is critical for epithelial-mesenchymal transition and stemness and promotes pancreatic carcinogenesis. Mol Cancer 10:8CrossRefGoogle Scholar
  19. Hershfield MS, Ganson NJ, Kelly SJ et al (2014) Induced and pre-existing anti-polyethylene glycol antibody in a trial of every 3-week dosing of pegloticase for refractory gout, including in organ transplant recipients. Arthritis Res Ther 16:R63CrossRefGoogle Scholar
  20. Holloway SE, Beck AW, Girard L et al (2005) Increased expression of Cyr61 (CCN1) identified in peritoneal metastases from human pancreatic cancer. J Am Coll Surg 200:371–377CrossRefGoogle Scholar
  21. Jevsevar S, Kunstelj M, Porekar VG (2010) PEGylation of therapeutic proteins. Biotechnol J 5:113–128CrossRefGoogle Scholar
  22. Jiang WG, Watkins G, Fodstad O et al (2004) Differential expression of the CCN family members Cyr61, CTGF and Nov in human breast cancer. Endocr Relat Cancer 11:781–791CrossRefGoogle Scholar
  23. Johnson SK, Stewart JP, Bam R et al (2014) CYR61/CCN1 overexpression in the myeloma microenvironment is associated with superior survival and reduced bone disease. Blood 124:2051–2060CrossRefGoogle Scholar
  24. Johnston E, Crawford J, Blackwell S et al (2000) Randomized, dose-escalation study of SD/01 compared with daily filgrastim in patients receiving chemotherapy. J Clin Oncol 18:2522–2528CrossRefGoogle Scholar
  25. Kim C, Axup J, Schultz P (2013) Protein conjugation with genetically encoded unnatural amino acids. Curr Opin Chem Biol 17:412–419CrossRefGoogle Scholar
  26. Knop K, Hoogenboom R, Fischer D et al (2010) Poly(ethylene glycol) in drug delivery: pros and cons as well as potential alternatives. Angew Chem 49:6288–6308CrossRefGoogle Scholar
  27. Kozlowski A, Harris JM (2001) Improvements in protein PEGylation: pegylated interferons for treatment of hepatitis C. Journal of controlled release : official journal of the Controlled Release Society 72:217–224CrossRefGoogle Scholar
  28. Lang K, Chin J (2014) Cellular incorporation of unnatural amino acids and bioorthogonal labeling of proteins. Chem Rev 114:4764–4806CrossRefGoogle Scholar
  29. Leask A (2011) CCN1: a novel target for pancreatic cancer. Journal of cell communication and signaling 5:123–124CrossRefGoogle Scholar
  30. Leask A (2013) Sonic advance: CCN1 regulates sonic hedgehog in pancreatic cancer. Journal of cell communication and signaling 7:61–62CrossRefGoogle Scholar
  31. Luxenhofer R, Bezen M, Jotdan R (2008) Kinetic investigations on the polymerization of 2-Oxazolines using Pluritriflate Initators. Macromolecular Rapid Communication 29:1509–1513CrossRefGoogle Scholar
  32. Macdougall IC (2005) CERA (continuous erythropoietin receptor activator): a new erythropoiesis-stimulating agent for the treatment of anemia. Curr Hematol Rep 4:436–440PubMedGoogle Scholar
  33. Macdougall IC, Roche A (2005) Administration of intravenous iron sucrose as a 2-minute push to CKD patients: a prospective evaluation of 2,297 injections. Am J Kidney Dis 46:283–289CrossRefGoogle Scholar
  34. Milla P, Dosio F, Cattel L (2012) PEGylation of proteins and liposomes: a powerful and flexible strategy to improve the drug delivery. Curr Drug Metab 13:105–119CrossRefGoogle Scholar
  35. Mishra, P.N., B.; Dey, R.K. (2016). PEGylation in anti-cancer therapy: an overview. Asian Journal of Pharmaceutical Sciences 11, 337–348CrossRefGoogle Scholar
  36. Ning X, Guo J, Wolfert M et al (2008) Visualizing metabolically labeled glycoconjugates of living cells by copper-free and fast huisgen cycloadditions. Angewandte Chemie-International Edition 47:2253–2255CrossRefGoogle Scholar
  37. Nischan N, Hackenberger CP (2014) Site-specific PEGylation of proteins: recent developments. J Org Chem 79:10727–10733CrossRefGoogle Scholar
  38. Nucci R, Raia CA, Vaccaro C et al (1991) Allosteric modifier and substrate binding of donkey deoxycytidylate aminohydrolase (EC 3.5.4.12). Arch Biochem Biophys 289:19–25CrossRefGoogle Scholar
  39. O'Kelly JK, Koeffler HP (2005) The role of CCN1 in tumorigenesis and cancer progression. In: Perbal BT (ed) In CCN proteins: anew family of cell growth and differentiation regulators. Imperial College Press, London, pp 273–291CrossRefGoogle Scholar
  40. Oelmeier SA, Dismer F, Hubbuch J (2012) Molecular dynamics simulations on aqueous two-phase systems - single PEG-molecules in solution. BMC Biophys 5:14CrossRefGoogle Scholar
  41. Oh Y, Swierczewska M, Kim TH et al (2015) Delivery of tumor-homing TRAIL sensitizer with long-acting TRAIL as a therapy for TRAIL-resistant tumors. Journal of controlled release : official journal of the Controlled Release Society 220:671–681CrossRefGoogle Scholar
  42. Pasut G, Canal F, Dalla Via L et al (2008) Antitumoral activity of PEG-gemcitabine prodrugs targeted by folic acid. Journal of controlled release : official journal of the Controlled Release Society 127:239–248CrossRefGoogle Scholar
  43. Pasut G, Greco F, Mero A et al (2009) Polymer-drug conjugates for combination anticancer therapy: investigating the mechanism of action. J Med Chem 52:6499–6502CrossRefGoogle Scholar
  44. Pasut G, Veronese F (2007) Polymer-drug conjugation, recent achievements and general strategies. Prog Polym Sci 32:933–961CrossRefGoogle Scholar
  45. Pepinsky RB, LePage DJ, Gill A et al (2001) Improved pharmacokinetic properties of a polyethylene glycol-modified form of interferon-beta-1a with preserved in vitro bioactivity. J Pharmacol Exp Ther 297:1059–1066PubMedGoogle Scholar
  46. Pfister D, Morbidelli M (2014) Process for protein PEGylation. Journal of controlled release : official journal of the Controlled Release Society 180:134–149CrossRefGoogle Scholar
  47. Pisal DS, Kosloski MP, Balu-Iyer SV (2010) Delivery of therapeutic proteins. J Pharm Sci 99:2557–2575CrossRefGoogle Scholar
  48. Qian X, Dong H, Tian H et al (2013) Characterization of a site-specific PEGylated analog of exendin-4 and determination of the PEGylation site. Int J Pharm 454:553–558CrossRefGoogle Scholar
  49. Roberts MJ, Bentley MD, Harris JM (2002) Chemistry for peptide and protein PEGylation. Adv Drug Deliv Rev 54:459–476CrossRefGoogle Scholar
  50. Su YC, Burnouf PA, Chuang KH et al (2017) Conditional internalization of PEGylated nanomedicines by PEG engagers for triple negative breast cancer therapy. Nat Commun 8:15507CrossRefGoogle Scholar
  51. Swierczewska M, Han HS, Kim K et al (2016) Polysaccharide-based nanoparticles for theranostic nanomedicine. Adv Drug Deliv Rev 99:70–84CrossRefGoogle Scholar
  52. Swierczewska M, Lee KC, Lee S (2015) What is the future of PEGylated therapies? Expert Opin Emerg Drugs 20:531–536CrossRefGoogle Scholar
  53. Thorner MO (1999) The discovery of growth hormone-releasing hormone. J Clin Endocrinol Metab 84:4671–4676PubMedGoogle Scholar
  54. Thorner MO, Strasburger CJ, Wu Z et al (1999) Growth hormone (GH) receptor blockade with a PEG-modified GH (B2036-PEG) lowers serum insulin-like growth factor-I but does not acutely stimulate serum GH. J Clin Endocrinol Metab 84:2098–2103PubMedGoogle Scholar
  55. Torchilin VP (2007) Targeted pharmaceutical nanocarriers for cancer therapy and imaging. AAPS J 9:E128–E147CrossRefGoogle Scholar
  56. van Geel R, Pruijn G, van Delft F et al (2012) Preventing thiol-Yne addition improves the specificity of strain-promoted Azide-alkyne cycloaddition. Bioconjug Chem 23:392–398CrossRefGoogle Scholar
  57. Veronese FM (2001) Peptide and protein PEGylation: a review of problems and solutions. Biomaterials 22:405–417CrossRefGoogle Scholar
  58. Veronese FM, Sacca B, Polverino de Laureto P et al (2001) New PEGs for peptide and protein modification, suitable for identification of the PEGylation site. Bioconjug Chem 12:62–70CrossRefGoogle Scholar
  59. Veronese FM, Schiavon O, Pasut G et al (2005) PEG-doxorubicin conjugates: influence of polymer structure on drug release, in vitro cytotoxicity, biodistribution, and antitumor activity. Bioconjug Chem 16:775–784CrossRefGoogle Scholar
  60. Wang YS, Youngster S, Bausch J et al (2000) Identification of the major positional isomer of pegylated interferon alpha-2b. Biochemistry 39:10634–10640CrossRefGoogle Scholar
  61. Yang JB, Duan ZJ, Yao W et al (2001) Synergistic transcriptional activation of human acyl-coenzyme a: cholesterol acyltransterase-1 gene by interferon-gamma and all-trans-retinoic acid THP-1 cells. J Biol Chem 276:20989–20998CrossRefGoogle Scholar
  62. Zalipsky S (1995) Functionalized poly(ethylene glycol) for preparation of biologically relevant conjugates. Bioconjug Chem 6:150–165CrossRefGoogle Scholar
  63. Zalipsky S, Qazen M, Walker JA 2nd et al (1999) New detachable poly(ethylene glycol) conjugates: cysteine-cleavable lipopolymers regenerating natural phospholipid, diacyl phosphatidylethanolamine. Bioconjug Chem 10:703–707CrossRefGoogle Scholar

Copyright information

© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2018

Authors and Affiliations

  • Vijayalaxmi Gupta
    • 1
    • 2
  • Sneha Bhavanasi
    • 1
  • Mohiuddin Quadir
    • 3
  • Kevin Singh
    • 1
  • Gaurav Ghosh
    • 1
  • Kritin Vasamreddy
    • 1
  • Arnab Ghosh
    • 1
    • 4
  • Teruna J. Siahaan
    • 5
  • Snigdha Banerjee
    • 1
    • 4
  • Sushanta K. Banerjee
    • 1
    • 4
  1. 1.Cancer Research UnitVA Medical CenterKansas CityUSA
  2. 2.Department of Obstetrics and GynecologyUniversity of Kansas Medical CenterKansas CityUSA
  3. 3.Department of Coatings and Polymeric MaterialsNorth Dakota State UniversityFargoUSA
  4. 4.Department of Pathology and Laboratory MedicineUniversity of Kansas Medical CenterKansas CityUSA
  5. 5.School of Pharmacy-Pharmaceutical ChemistryThe University of KansasLawrenceUSA

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