Synthetic self-adjuvanted multivalent Mucin 1 (MUC1) glycopeptide vaccines with improved in vivo antitumor efficacy

Abstract

The tumor-associated glycoprotein Mucin 1 (MUC1) is aberrantly glycosylated on cancer cells and is considered a promising target for antitumor vaccines. The weak immunogenicity and low sequence homology of mouse mucins and human MUC1 are the main obstacles for the development of vaccines. Herein, a self-adjuvanted strategy combining toll-like receptor 2 lipopeptide ligands and T-cell epitopes and the multivalent effect were used to amplify the immune response and evade the unpredictable immunogenicity, generating two self-adjuvanted three-component MUC1 vaccines (mono- and trivalent MUC1 vaccines). To simulate the aberrantly glycosylated MUC1 glycoprotein, the MUC1 tandem repeat peptide was bounded with Tn antigens at T9, S15, and T16, and served as B-cell epitopes. Results showed that both vaccines elicited a robust antibody response in wild-type mice compared with a weaker response in MUC1 transgenic mice. The trivalent vaccine did not elevate the antibody response level compared with the monovalent vaccine; however, a more delayed tumor growth and prolonged survival time was realized in wild-type and transgenic mouse models treated with the trivalent vaccine. These results indicate that the self-adjuvanted three-component MUC1 vaccines, especially the trivalent vaccine, can trigger robust antitumor effects regardless of sequence homology, and, therefore, show promise for clinical translation.

Keywords

cancer vaccine / MUC1 glycopeptide / multivalent strategy / self-adjuvanted vaccine

References

[1] Joshi S, Kumar S, Bafna S, et al. Genetically engineered mucin mouse models for inflammation and cancer. Cancer Metastasis Rev. 2015;34(4):593-609.

[2] Spicer AP, Parry G, Patton S, et al. Molecular cloning and analysis of the mouse homologue of the tumor-associated mucin, MUC1, reveals conservation of potential O-glycosylation sites, transmembrane, and cytoplasmic domains and a loss of minisatellite-like polymorphism. J Biol Chem. 1991;266(23):15099-15109.

[3] Fang T, Van Elssen C, Duarte JN, et al. Targeted antigen delivery by an anti-class II MHC VHH elicits focused alphaMUC1(Tn) immunity. Chem Sci. 2017;8(8):5591-5597.

[4] Ju T, Lanneau GS, Gautam T, et al. Human tumor antigens Tn and sialyl Tn arise from mutations in Cosmc. Cancer Res. 2008;68(6):1636-1646.

[5] Radhakrishnan P, Dabelsteen S, Madsen FB, et al. Immature truncated O-glycophenotype of cancer directly induces oncogenic features. Proc Natl Acad Sci USA. 2014;111(39):E4066-E4075.

[6] Sanz-Martinez I, Pereira S, Merino P, et al. Molecular recognition of GalNAc in mucin-type O-glycosylation. Acc Chem Res. 2023;56(5):548-560.

[7] Lakshminarayanan V, Thompson P, Wolfert MA, et al. Immune recognition of tumor-associated mucin MUC1 is achieved by a fully synthetic aberrantly glycosylated MUC1 tripartite vaccine. Proc Natl Acad Sci USA. 2012;109(1):261-266.

[8] Gaidzik N, Westerlind U, Kunz H. The development of synthetic antitumour vaccines from mucinglycopeptide antigens. Chem Soc Rev. 2013;42(10):4421-4442.

[9] Keil S, Claus C, Dippold W, et al. Towards the development of antitumor vaccines: a synthetic conjugate of a tumor-associated MUC1 glycopeptide antigen and a tetanus toxin epitope. Angew Chem Int Ed. 2001;40(2):366-369.

[10] Dziadek S, Kowalczyk D, Kunz H. Synthetic vaccines consisting of tumor-associated MUC1 glycopeptide antigens and bovine serum albumin. Angew Chem Int Ed Engl. 2005;44(46):7624-7630.

[11] Huang ZH, Shi L, Ma JW, et al. A totally synthetic, self-assembling, adjuvant-free MUC1 glycopeptide vaccine for cancer therapy. J Am Chem Soc. 2012;134(21):8730-8733.

[12] Pifferi C, Aguinagalde L, Ruiz-de-Angulo A, et al. Development of synthetic, self-adjuvanting, and self-assembling anticancer vaccines based on a minimal saponin adjuvant and the tumor-associated MUC1 antigen. Chem Sci. 2023;14(13):3501-3513.

[13] Wilkinson BL, Day S, Malins LR, et al. Self-adjuvanting multicomponent cancer vaccine candidates combining per-glycosylated MUC1 glycopeptides and the Toll-like receptor 2 agonist Pam₃CysSer. Angew Chem Int Ed Engl. 2011;50(7):1635-1639.

[14] Gabba A, Attariya R, Behren S, et al. MUC1 glycopeptide vaccine modified with a GalNAc glycocluster targets the macrophage galactose C-type lectin on dendritic cells to elicit an improved humoral response. J Am Chem Soc. 2023;145(24):13027-13037.

[15] Palitzsch B, Hartmann S, Stergiou N, et al. A fully synthetic four-component antitumor vaccine consisting of a mucin glycopeptide antigen combined with three different T-helper-cell epitopes. Angew Chem Int Ed Engl. 2014;53(51):14245-14249.

[16] Kaiser A, Gaidzik N, Becker T, et al. Fully synthetic vaccines consisting of tumor-associated MUC1 glycopeptides and a lipopeptide ligand of the Toll-like receptor 2. Angew Chem Int Ed Engl. 2010;49(21):3688-3692.

[17] Apostolopoulos V, Osinski C, Mckenzie I. MUC1 cross-reactive Galα(l,3)Gal antibodies in humans switch immune responses from cellular to humoral. Nat Med. 1998;4:315-320.

[18] Kaiser A, Gaidzik N, Westerlind U, et al. A synthetic vaccine consisting of a tumor-associated sialyl-T(N)-MUC1 tandem-repeat glycopeptide and tetanus toxoid: induction of a strong and highly selective immune response. Angew Chem Int Ed Engl. 2009;48(41):7551-7555.

[19] Westerlind U, Hobel A, Gaidzik N, et al. Synthetic vaccines consisting of tumor-associated MUC1 glycopeptide antigens and a T-cell epitope for the induction of a highly specific humoral immune response. Angew Chem Int Ed Engl. 2008;47(39):7551-7556.

[20] Palitzsch B, Gaidzik N, Stergiou N, et al. A synthetic glycopeptide vaccine for the induction of a monoclonal antibody that differentiates between normal and tumor mammary cells and enables the diagnosis of human pancreatic cancer. Angew Chem Int Ed Engl. 2016;55(8):2894-2898.

[21] Martinez-Saez N, Peregrina JM, Corzana F. Principles of mucin structure: implications for the rational design of cancer vaccines derived from MUC1-glycopeptides. Chem Soc Rev. 2017;46(23):7154-7175.

[22] Coelho H, Matsushita T, Artigas G, et al. The quest for anticancer vaccines: deciphering the fine-epitope specificity of cancer-related monoclonal antibodies by combining microarray screening and saturation transfer difference NMR. J Am Chem Soc. 2015;137(39):12438-12441.

[23] Cai H, Sun ZY, Chen MS, et al. Synthetic multivalent glycopeptide-lipopeptide antitumor vaccines: impact of the cluster effect on the killing of tumor cells. Angew Chem Int Ed Engl. 2014;53(6):1699-1703.

[24] Martinez-Saez N, Supekar NT, Wolfert MA, et al. Mucin architecture behind the immune response: design, evaluation and conformational analysis of an antitumor vaccine derived from an unnatural MUC1 fragment. Chem Sci. 2016;7(3):2294-2301.

[25] Braun P, Davies GM, Price MR, et al. Effects of glycosylation on fragments of tumour associated human epithelial mucin MUC1. Bioorg Med Chem. 1998;6(9):1531-1545.

[26] Fasting C, Schalley CA, Weber M, et al. Multivalency as a chemical organization and action principle. Angew Chem Int Ed Engl. 2012;51(42):10472-10498.

[27] Ryan SO, Turner MS, Gariepy J, et al. Tumor antigen epitopes interpreted by the immune system as self or abnormal-self differentially affect cancer vaccine responses. Cancer Res. 2010;70(14):5788-5796.

[28] Cai H, Zhang R, Orwenyo J, et al. Multivalent antigen presentation enhances the immunogenicity of a synthetic three-component HIV-1 V3 glycopeptide vaccine. ACS Cent Sci. 2018;4(5):582-589.

[29] Tirand L, Frochot C, Vanderesse R, et al. A peptide competing with VEGF165 binding on neuropilin-1 mediates targeting of a chlorin-type photosensitizer and potentiates its photodynamic activity in human endothelial cells. J Control Release. 2006;111(1-2):153-164.

[30] Lei H, Shen Y, Song L, et al. Hapten synthesis and antibody production for the development of a melamine immunoassay. Anal Chim Acta. 2010;665(1):84-90.

[31] Paolino D, Licciardi M, Celia C, et al. Bisphosphonate-polyaspartamide conjugates as bone targeted drug delivery systems. J Mater Chem B. 2015;3(2):250-259.

[32] Cai H, Huang ZH, Shi L, et al. Synthesis of Tn/T antigen MUC1 glycopeptide BSA conjugates and their evaluation as vaccines. Eur J Org Chem. 2011;2011(20-21):3685-3689.

[33] Zhou SH, Li YT, Zhang RY, et al. Alum adjuvant and built-in TLR7 agonist synergistically enhance anti-MUC1 immune responses for cancer vaccine. Front Immunol. 2022;13:857779.

[34] Yu J, Song Y, Tian W. How to select IgG subclasses in developing anti-tumor therapeutic antibodies. J Hematol Oncol. 2020;13(1):45.

[35] Cabral MP, Garcia P, Beceiro A, et al. Design of live attenuated bacterial vaccines based on D-glutamate auxotrophy. Nat Commun. 2017;8:15480.

[36] Collins AM. IgG subclass co-expression brings harmony to the quartet model of murine IgG function. Immunol Cell Biol. 2016;94(10):949-954.

[37] Du JJ, Zou SY, Chen XZ, et al. Liposomal antitumor vaccines targeting mucin 1 elicit a lipid-dependent immunodominant response. Chem Asian J. 2019;14(12):2116-2121.

[38] Yi M, Niu M, Xu L, et al. Regulation of PD-L1 expression in the tumor microenvironment. J Hematol Oncol. 2021;14(1):10.

[39] Pan J, Zeng W, Jia J, et al. A novel therapeutic tumor vaccine targeting MUC1 in combination with PD-L1 elicits specific anti-tumor immunity in mice. Vaccines (Basel). 2022;10(7):1092.

[40] Dai J, Hu JJ, Dong X, et al. Deep downregulation of PD-L1 by caged peptide-conjugated AIEgen/miR-140 nanoparticles for enhanced immunotherapy. Angew Chem Int Ed Engl. 2022;61(18):e202117798.

[41] Bouillez A, Adeegbe D, Jin C, et al. MUC1-C promotes the suppressive immune microenvironment in non-small cell lung cancer. Oncoimmunology. 2017;6(9):e1338998.

[42] Nuhn L, Hartmann S, Palitzsch B, et al. Water-soluble polymers coupled with glycopeptide antigens and T-cell epitopes as potential antitumor vaccines. Angew Chem Int Ed Engl. 2013;52(40):10652-10656.

[43] Cai H, Chen MS, Sun ZY, et al. Self-adjuvanting synthetic antitumor vaccines from MUC1 glycopeptides conjugated to T-cell epitopes from tetanus toxoid. Angew Chem Int Ed Engl. 2013;52(23):6106-6110.

[44] Pett C, Cai H, Liu J, et al. Microarray analysis of antibodies induced with synthetic antitumor vaccines: specificity against diverse mucin core structures. Chem. 2017;23(16):3875-3884.

[45] Yan S, Rolfe BE, Zhang B, et al. Polarized immune responses modulated by layered double hydroxides nanoparticle conjugated with CpG. Biomaterials. 2014;35(35):9508-9516.

[46] Liu GH, Chen T, Zhang X, et al. Small molecule inhibitors targeting the cancers. MedComm. 2022;3(4):e181.

[47] Hou Y, Yan T, Cao H, et al. Chimeric hepatitis B virus core particles displaying Neisserial surface protein A confer protection against virulent Neisseria meningitidis serogroup B in BALB/c mice. Int J Nanomed. 2019;14:6601-6613.

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