Engineered asparaginase from Erwinia chrysanthemi enhances asparagine hydrolase activity and diminishes enzyme immunoreactivity ‐ a new promise to treat acute lymphoblastic leukemia
| العنوان: | Engineered asparaginase from |
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| المؤلفون: | Guilherme Meira Lima, Gisele Monteiro, Débora Custodio Moura, Camila O. dos Santos, Marcos Antonio de Oliveira, Iris Munhoz Costa, Adalberto Pessoa |
| المساهمون: | Universidade de São Paulo (USP), Cold Spring Harbor, Universidade Estadual Paulista (UNESP) |
| المصدر: | Scopus Repositório Institucional da UNESP Universidade Estadual Paulista (UNESP) instacron:UNESP |
| بيانات النشر: | Wiley, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | immunogenic epitopes, lower therapeutic ASNase dose, Asparaginase, l-asparaginase, General Chemical Engineering, Pharmacology, Epitope, Inorganic Chemistry, chemistry.chemical_compound, medicine, Erwinia chrysanthemi, Asparagine, Waste Management and Disposal, chemistry.chemical_classification, biology, Renewable Energy, Sustainability and the Environment, Chemistry, Organic Chemistry, protein engineering, medicine.disease, Pollution, Enzyme assay, Leukemia, Fuel Technology, Enzyme, Toxicity, biology.protein, Specific activity, mutation, Biotechnology |
| الوصف: | Made available in DSpace on 2022-04-28T19:45:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) BACKGROUND: The treatment of acute lymphoblastic leukemia (ALL) uses the biopharmaceutical l-asparaginase (ASNase) as the main medication. This drug, from bacterial origin (Escherichia coli or Erwinia chrysanthemi), depletes l-asparagine (Asn) and secondarily l-glutamine (Gln – GLNase activity) from the bloodstream, leading leukemic cells to die by deprivation of Asn. The use of ASNase is limited by the high incidence of adverse effects, which collectively can specifically impair quality of life of patients. Its high toxicity caused by the product of the hydrolysis of amino acids and the formation of anti-ASNase antibodies often required treatment interruption, thus reducing the chances of cure and increasing the rates of disease relapse. RESULTS: In order to improve enzymatic activity, while reducing toxicity, we developed through directed evolution a double-mutant ASNase from Erwinia chrysanthemi (Erw_DM), which has specific activity for Asn 46% higher than the wild-type enzyme (Erw_WT). This makes it possible to reduce the amount of protein necessary for depletion of this amino acid and, consequently, the reduction of GLNase activity, considered toxic. In silico analysis showed that a lower number of epitopes was exposed, resulting in reduced recognition of the recombinant protein by antibody anti-ASNase observed in vitro assay. Furthermore, we observed the same cytotoxic profile for the MOLT-4 and REH ALL cell lines using 40% lower protein mass of Erw_DM to achieve the minimum enzyme activity required in the bloodstream during treatment. CONCLUSION: Altogether, our findings describe a potent and less immunogenic ASNase, an improvement that may alleviate treatment adverse effects developed in anti-ALL therapy. © 2021 Society of Chemical Industry (SCI). Departamento de Tecnologia Bioquímico-Farmacêutica Faculdade de Ciências Farmacêuticas Universidade de São Paulo Cold Spring Harbor Laboratory Cold Spring Harbor Instituto de Biociências Universidade Estadual Paulista Instituto de Biociências Universidade Estadual Paulista CAPES: 001 FAPESP: 2013/08617-7 FAPESP: 2015/07749-2 FAPESP: 2016/25896-5 FAPESP: 2018/15104-0 CNPq: 309224/2019-5 |
| تدمد: | 1097-4660 0268-2575 |
| DOI: | 10.1002/jctb.6933 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::70a615a6d0d195575438c2324c30fc47 https://doi.org/10.1002/jctb.6933 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....70a615a6d0d195575438c2324c30fc47 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10974660 02682575 |
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| DOI: | 10.1002/jctb.6933 |