Influence of lysosomal protease sensitivity in the immunogenicity of the antitumor biopharmaceutical asparaginase

Author(s):Show less -

Rodrigues, Mariane A. D. ^[1] ; Pimenta, Marcela V. ^[1] ; Costa, Iris M. ^[1] ; Zenatti, Priscila P. ^[2] ; Migita, Natacha A. ^{[2, 3]} ; Yunes, Jose A. ^{[2, 3]} ; Rangel-Yagui, Carlota O. ^[1] ; de Sa, Matheus M. ^[4] ; Pessoa, Adalberto ^[1] ; Costa-Silva, Tales A. ^[1] ; Toyama, Marcos H. ^[5] ; Breyer, Carlos A. ^[5] ; de Oliveira, Marcos A. ^[5] ; Santiago, Veronica F. ^[6] ; Palmisano, Giuseppe ^[6] ; Barbosa, Christiano M. V. ^[7] ; Hebeda, Cristina B. ^[7] ; Farsky, Sandra H. P. ^[7] ; Monteiro, Gisele ^[1]

Affiliation

^[1] Univ Sao Paulo, Dept Tecnol Bioquim Farmaceut, Fac Ciencias Farmaceut, Sao Paulo - Brazil

^[2] Ctr Infantil Boldrini, Sao Paulo - Brazil

^[3] Univ Estadual Campinas, Fac Med Sci, Dept Med Genet, Sao Paulo - Brazil

^[4] Univ Sao Paulo, Heart Inst InCor, Sch Med, Sao Paulo - Brazil

^[5] UNESP Sao Paulo State Univ, Biosci Inst, Coastal Campus, Sao Paulo - Brazil

^[6] Univ Sao Paulo, Inst Biomed Sci, Dept Parasitol, Sao Paulo - Brazil

^[7] Univ Sao Paulo, Fac Ciencias Farmaceut, Dept Clin & Toxicol Anal, Sao Paulo - Brazil

• Document type: Journal article
• Source: Biochemical Pharmacology; v. 182, DEC 2020.
• Web of Science Citations: 0

Abstract

L-asparaginase (ASNase) from Escherichia coli (EcAII) is used in the treatment of acute lymphoblastic leukaemia (ALL). EcAII activity in vivo has been described to be influenced by the human lysosomal proteases asparaginyl endopeptidase (AEP) and cathepsin B (CTSB); these hydrolases cleave and could expose epitopes associated with the immune response against EcAII. In this work, we show that ASNase resistance to CTSB and/or AEP influences the formation of anti-ASNase antibodies, one of the main causes of hypersensitivity reactions in patients. Error-prone polymerase chain reaction was used to produce variants of EcAII more resistant to proteolytic cleavage by AEP and CTSB. The variants with enzymatic activity and cytotoxicity levels equivalent to or better than EcAII WT were submitted to in vivo assays. Only one of the mutants presented increased serum half-life, so resistance to these proteases is not the only feature involved in EcAII stability in vivo. Our results showed alteration of the phenotypic profile of B cells isolated after animal treatment with different protease-resistant proteoforms. Furthermore, mice that were exposed to the protease-resistant proteoforms presented lower anti-asparaginase antibodies production in vivo. Our data suggest that modulating resistance to lysosomal proteases can result in less immunogenic protein drugs. (AU)

• FAPESP's process: 14/06863-3 - Post-translational modifications in cancer and parasite infection diagnosis: methodological approaches and biological implications
• Grantee: Giuseppe Palmisano
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 18/18257-1 - Multi-user equipment approved in grant 14/06863-3: HPLC system configured for analysis of carbohydrates, amino acidis, peptides and glycoproteins
• Grantee: Giuseppe Palmisano
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 13/08617-7 - Production of extracellular L-asparaginase: from bioprospecting to the engineering of an antileukemic biopharmaceutical
• Grantee: Adalberto Pessoa Junior
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 16/25896-5 - Biochemical characterization and cytotoxic evaluation of mutant isoforms of L-Asparaginase II from Dickeya chrysanthemi (Erwinia chrysanthemi)
• Grantee: Iris Munhoz Costa
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 15/07749-2 - Protein engineering and comparison of microbial expression systems of the biopharmaceutical L-asparaginase
• Grantee: Gisele Monteiro
• Support Opportunities: Regular Research Grants
• FAPESP's process: 18/15104-0 - Preclinical assays of glycoprotein asparaginase proteoforms or resistant to serum proteases.
• Grantee: Gisele Monteiro
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/08139-8 - Cloning, expression and charactherization of L-asparaginases from Saccharomyces cerevisiae and comparison with bacterial L-asparaginases used to treat Leukemia
• Grantee: Mariana Silva Moreira Leite
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 18/15549-1 - Post-translational modifications in Chagas Disease biological processes and diagnostics: novel methodological approaches and biological applications
• Grantee: Giuseppe Palmisano
• Support Opportunities: Research Grants - Young Investigators Grants - Phase 2