Author(s):
Gibadullin, Ruslan ; Suárez, Óscar ; Lazaris, Foivos S. ; Gutiez, Naiara ; Atondo, Estibaliz ; Araujo-Aris, Sarai ; Eguskiza, Ander ; Niu, Jiani ; Kuhn, Ariel J. ; Grosso, Ana S. ; Rodriguez, Héctor ; García-Martín, Fayna ; Marcelo, Filipa ; Santos, Tanausú ; Avenoza, Alberto ; Busto, Jesús H. ; Peregrina, Jesús M. ; Gellman, Samuel H. ; Anguita, Juan ; Fiammengo, Roberto ; Corzana, Francisco
Date: 2025
Persistent ID: http://hdl.handle.net/10362/185586
Origin: Repositório Institucional da UNL
Project/scholarship:
info:eu-repo/grantAgreement/EC/H2020/956544/EU;
info:eu-repo/grantAgreement/FCT/Concurso para Financiamento de Projetos de Investigação Científica e Desenvolvimento Tecnológico em Todos os Domínios Científicos - 2020/PTDC%2FQUI-OUT%2F2586%2F2020/PT;
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT;
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT;
info:eu-repo/grantAgreement/FCT/Concurso para Atribuição do Estatuto e Financiamento de Laboratórios Associados (LA)/LA%2FP%2F0140%2F2020/PT;
Subject(s): Cancer vaccine; Glycopeptide; Gold nanoparticles; Mucin; Proteolytic stability; β-peptides; Analytical Chemistry; Chemistry (miscellaneous); Physical and Theoretical Chemistry; Organic Chemistry; SDG 3 - Good Health and Well-being
Description
Funding Information: This project has received partial funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 956544. This work was also supported in part by the National Institutes of Health grant (R01 GM056414 and its successor R35 GM151985) awarded to S.H.G. We thank the Agencia Estatal de Investigación (AEI, PID2021-127622OB-I00 to F.C., J.H.B. and O.S., PDC2022-133725-C21 to F.C, J.H.B., PID2020-120099RA-I00 to F.G.M., PID2021-124328OB-100 to J.A., and PRE-2019-091720 to S.A.), Universidad de La Rioja (REGI22/47 and REGI22/16) and Asociación Española contra el Cáncer (AECC) sección La Rioja (F.C. and J.M.P.). F.M. and A.S.G. acknowledge Fundação para a Ciência e Tecnologia Portugal (FCT-Portugal) for the MGL4Life project 10.54499/PTDC/QUI-OUT/2586/2020. F.M. thanks FCT-Portugal for CEECINST/00042/2021/CP1773/CT0011. F.M and A.S.G. thank UCIBIO project (UIDP/04378/2020 and UIDB/04378/2020), and Associate Laboratory Institute for Health and Bioeconomy - i4HB project (LA/P/0140/2020) and the National NMR Facility supported by FCT-Portugal (ROTEIRO/0031/2013–PINFRA/22161/2016, cofinanced by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). T.S. thanks MCIN/AEI/10.13039/501100011033 and the European Union NextGenerationEU/PRTR for his Juan de la Cierva contract, JDC2022-048607-I. N.G. held a predoctoral grant from the Scientific Foundation of the Spanish Association Against Cancer in Bizkaia (PRDVZ222452GUTI). CICbioGUNE is the recipient of a Severo Ochoa Centro de Excelencia Award (CEX2021-001136-S). Supported in part by Fundación Jesús de Gangoiti. We are also grateful for partial support from the Vilas Trust and from the University of Wisconsin-Madison Office of the Vice Chancellor for Research and Graduate Education with funding from the Wisconsin Alumni Research Foundation. A.J.K. was supported in part by a postdoctoral fellowship from the National Institutes of Health (5F32AI176876). Publisher Copyright: © 2025 The Authors. Published by American Chemical Society.
Glycopeptides derived from the mucin-1 (MUC1) glycoprotein hold significant promise as cancer vaccine candidates, but their clinical utility is limited by proteolytic degradation and the poor bioavailability of L-α-amino acid-based peptides. In this study, we demonstrate that substitution of multiple α-amino acids with homologous β-amino acids (same side chain, but extended backbone) in O-glycosylated MUC1 derivatives significantly enhances their proteolytic stability. We further show that α-to-β substitutions within the most immunogenic epitope of MUC1 impede binding to an anti-MUC1 antibody, while substitutions outside the same epitope preserve antibody recognition. Structural investigations using circular dichroism, NMR spectroscopy, and molecular dynamics simulations reveal that the strongest α/β-peptide binders retain native-like conformations in the epitope region, both in their unbound state and when bound to the anti-MUC1 antibody. Conjugation of these high-affinity α/β-peptide analogs to gold nanoparticles induces robust immune responses in mice comparable to that of the native glycopeptide. Additionally, these α/β-analogs elicit elevated levels of the cytokine IFNγ, one of the key proteins for tumor cell elimination, surpassing levels produced by the native MUC1 glycopeptide. In contrast, a low-affinity α/β-analogue with lower proteolytic stability produces minimal cytokine responses, underscoring the critical role of these biochemical properties in vaccine efficacy. Collectively, our findings highlight that α-to-β modifications in the peptide backbone offer an effective strategy for developing biostable, highly immunogenic glycopeptide-based cancer vaccines, exemplifying the power of structure-based rational design in advancing next-generation vaccines.
