Designing a Humanized Immunotoxin Based on Gelonin Against HER2+ Breast Cancer: An In-Silico Study
DOI:
https://doi.org/10.1590/s2175-97902026e24193Keywords:
Breast cancer, Immunotoxin, Bioinformatics, HER2, GeloninAbstract
Recombinant immunotoxins (RITs) are considered tumor-specific therapeutics because their variable antibody fragment can specifically bind to target cells, and their engineered toxin fragment specifically kills cancerous cells upon internalization. Human epidermal growth factor receptor 2 (HER2) is a transmembrane tyrosine kinase receptor that is overexpressed in breast cancer (BC) patients and can be used for targeted therapies in the form of immunotoxins. The current study aimed to develop pertuzumab gelonin as a targeted immunotoxin for HER2-positive breast cancer cells using bioinformatics methods and to analyze its properties in silico. The Fab light and heavy chain of pertuzumab were linked to gelonin's ribosome-inactivating region (RIP) region. The amino acid sequence of the antibody chains was obtained from the Protein Data Bank Server (PDB), and the sequence of the RIP region of gelonin was obtained from the National Center for Biotechnology Information (NCBI). The immunotoxin's physicochemical properties, secondary, tertiary, and 3D structure, mRNA folding, allergenicity, solubility, and antigenicity were predicted from online servers, and protein-protein docking was also performed. The immunotoxin is stable, non-allergenic, and binds to HER2 receptors with high affinity. Based on our findings, it can be a good targeted therapy for HER2 breast cancer.
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Barkhordari F, Rismani E, Tabasinezhad M, Asgari S, Nematollahi L, Talebkhan Y. Computational analysis of fusion protein of anti-HER2 scFv and alpha luffin: A new immunotoxin protein for HER2 positive cancers. Braz J Pharm Sci. 2022;58.
Bera TK, Onda M, Kreitman RJ, Pastan I. An improved recombinant Fab-immunotoxin targeting CD22 expressing malignancies. Leuk Res. 2014 Oct 1;38(10):1224–9.
Berstad MEB, Cheung LH, Weyergang A. Production of recombinant gelonin using an automated liquid chromatography system. Toxins (Basel). 2020 Aug 1;12(8).
Bredin P, Walshe JM, Denduluri N. Systemic therapy for metastatic HER2-positive breast cancer. Semin Oncol. 2020 Oct 1;47(5):259–69.
Buchan DWA, Jones DT. The PSIPRED Protein Analysis Workbench: 20 years on. Nucleic Acids Res. 2019 Jul 1;47(W1):W402–7.
Ding G B, Wu G, Li B, Yang P, Li Z. High-yield expression in Escherichia coli, biophysical characterization, and biological evaluation of plant toxin gelonin. 3 Biotech. 2019 Jan 1;9(1).
Ding GB, Zhu C, Wang Q, Cao H, Li BC, Yang P, et al. Molecularly engineered tumor acidity-responsive plant toxin gelonin for safe and efficient cancer therapy. Bioact Mater. 2022 Dec 1;18:42-55.
Ganji M, Khalili S, Mard-Soltani M, Khalesi B, Karkhah A, Amani J. A Precisely Designed Immunotoxin Against VCAM1 Consisting of a Humanized Antibody Variable Domain Fused to Granzyme: An In Silico Approach. Int J Pept Res Ther. 2020 Mar 1;26(1):129–37.
Global Biodiversity Information Facility [Internet]. Vol. 41, Choice Reviews Online. 2022. p. 41-5289-41–5289. Available from: https://www.gbif.org/species/3054898.
Guo R, Yang Y, Zhang D, Du J, Zhu X, Liu Y, et al. A bispecific immunotoxin (IHPP) with a long half-life targeting HER2 and PDGFRβ exhibited improved efficacy against HER2-positive tumors in a mouse xenograft model. Int J Pharm. 2021 Jan 5;592.
Islam E. Development of chemokine CXCL12-dependent immunotoxin against small cell lung cancer using in silico approaches. Inform Med Unlocked. 2021 Jan 1;25.
Keshtvarz M, Mahboobi M, Kieliszek M, Miecznikowski A, Sedighian H, Rezaei M, et al. Engineering of cytolethal distending toxin b by its reducing immunogenicity and maintaining stability as a new drug candidate for tumor therapy; an in silico study. Toxins (Basel). 2021 Nov 1;13(11).
Ko J, Park H, Heo L, Seok C. GalaxyWEB server for protein structure prediction and refinement. Nucleic Acids Res. 2012 Jul;40(W1).
Kreutzfeldt J, Rozeboom B, Dey N, De P. The trastuzumab era: current and upcoming targeted HER2+ breast cancer therapies. Am J Cancer Res [Internet]. 2020;10(4):1045–67. Available from: www.ajcr.us/.
Le Du F, Diéras V, Curigliano G. The role of tyrosine kinase inhibitors in the treatment of HER2+ metastatic breast cancer. Eur J Cancer. 2021 Sep 1;154:175–89.
Liu Q, Zhang L, Ji X, Shin MC, Xie S, Pan B, et al. A self-assembly and stimuli-responsive fusion gelonin delivery system for tumor treatment. J Ind Eng Chem. 2020 Sep 25;89:409–15.
Macindoe G, Mavridis L, Venkatraman V, Devignes MD, Ritchie DW. HexServer: An FFT-based protein docking server powered by graphics processors. Nucleic Acids Res. 2010 May 5;38(SUPPL. 2).
Mahmoudi R, Dianat-Moghadam H, Poorebrahim M, Siapoush S, Poortahmasebi V, Salahlou R, et al. Recombinant immunotoxins development for HER2-based targeted cancer therapies. Cancer Cell Int. 2021 Dec 1;21(1).
Mercogliano MF, Bruni S, Mauro FL, Schillaci R. Emerging Targeted Therapies for HER2-Positive Breast Cancer. Cancers (Basel). 2023 Apr 1;15(7).
Moghadam ZM, Halabian R, Sedighian H, Behzadi E, Amani J, Fooladi AAI. Designing and analyzing the structure of DT-STXB fusion protein as an anti-tumor agent: An in silico approach. Iran J Pathol. 2019 Sep 1;14(4):305–12.
Movahedpour A, Ahmadi K, Taheri-Anganeh M, Amiri A, Ahmadi N, Khatami SH, et al. Designing a Humanized Immunotoxin Based on HER2 Specific scFv and DFF40 Toxin Against Breast Cancer: An In-Silico Study. Int J Pept Res Ther. 2022;28(5).
Nika L, Cuadrado-Castano S, Arunkumar GA, Grünwald-Gruber C, McMahon M, Koczka K, et al. An HER2-displaying virus-like particle vaccine protects from challenge with mammary carcinoma cells in a mouse model. Vaccines. 2019 Jun 1;7(2).
Nugent T, Cozzetto D, Jones DT. Evaluation of predictions in the CASP10 model refinement category. Proteins Struct Funct Bioinforma. 2014 Feb;82(SUPPL.2):98–111.
Olsen CE, Cheung LH, Weyergang A, Berg K, Vallera DA, Rosenblum MG, et al. Design, characterization, and evaluation of scFvCD133/rgelonin: A CD133-targeting recombinant immunotoxin for use in combination with photochemical internalization. J Clin Med. 2020 Jan 1;9(1).
Park T, Min KA, Cheong H, Moon C, Shin MC. Genetic engineering and characterisation of chlorotoxin-fused gelonin for enhanced glioblastoma therapy. J Drug Target. 2019 Oct 21;27(9):950–8.
Peng Y, Wu Z, Pang Z, Zhang L, Song D, Liu F, et al. Manufacture and evaluation of a HER2-positive breast cancer immunotoxin 4D5Fv-PE25. Microb Cell Fact. 2023 Dec 1;22(1).
Quintero-Hernández V, Juárez-González VR, Ortíz-León M, Sánchez R, Possani LD, Becerril B. The change of the scFv into the Fab format improves the stability and in vivo toxin neutralization capacity of recombinant antibodies. Mol Immunol. 2007 Feb;44(6):1307–15.
Samavarchi Tehrani SS, Gharibi S, Movahedpour A, Goodarzi G, Jamali Z, Khatami SH, et al. Design and evaluation of scFv-RTX-A as a novel immunotoxin for breast cancer treatment: an in silico approach. J Immunoass Immunochem. 2021;42(1):19–33.
Sefid F, Payandeh Z, Azamirad G, Baradaran B, Afjadi MN, Islami M, et al. Atezolizumab and granzyme B as immunotoxin against PD-L1 antigen; an in-silico study. Silico Pharmacol. 2021 Feb 15;9(1).
Srivastava S, Verma S, Kamthania M, Kaur R, Badyal RK, Saxena AK, et al. Structural Basis for Designing Multiepitope Vaccines Against COVID-19 Infection: In Silico Vaccine Design and Validation. JMIR Bioinforma Biotechnol. 2020;1(1).
Stoessel A, Groysbeck N, Guyot L, Barret L, Nominé Y, Nguekeu-Zebaze L, et al. Modular Conjugation of a Potent Anti-HER2 Immunotoxin Using Coassociating Peptides. Bioconjug Chem. 2020 Oct 21;31(10):2421–30.
Vafadar A, Taheri-Anganeh M, Movahedpour A, Jamali Z, Irajie C, Ghasemi Y, et al. In silico design and evaluation of scFv-CdtB as a novel immunotoxin for breast cancer treatment. Int J Cancer Manag. 2020 Jan 1;13(1).
Yan Y, Tao H, He J, Huang SY. The HDOCK server for integrated protein–protein docking. Nat Protoc. 2020 May 1;15(5):1829–52.
Ye F, Dewanjee S, Li Y, Jha NK, Chen ZS, Kumar A, et al. Advancements in clinical aspects of targeted therapy and immunotherapy in breast cancer. Mol Cancer. 2023 Jul 6;22(1):105.
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