Glycoengineering of AAV-delivered monoclonal antibodies yields increased ADCC activity
| Autor: | Sebastian P. Fuchs, Guangping Gao, James M. Termini, Ronald C. Desrosiers, José M. Martinez-Navio |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
viral reservoir lcsh:QH426-470 medicine.drug_class Genetic enhancement viruses adeno-associated virus Monoclonal antibody medicine.disease_cause Fucose Neutralization 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Western blot Genetics medicine fucosyltransferase 8 lcsh:QH573-671 Molecular Biology Adeno-associated virus Antibody-dependent cell-mediated cytotoxicity biology medicine.diagnostic_test lcsh:Cytology broadly neutralizing antibodies glycoengineering Molecular biology gene therapy lcsh:Genetics 030104 developmental biology chemistry 030220 oncology & carcinogenesis biology.protein Molecular Medicine Original Article immunotherapy Antibody antibody-dependent cellular cytotoxicity |
| Zdroj: | Molecular Therapy: Methods & Clinical Development, Vol 20, Iss, Pp 204-217 (2021) Molecular Therapy. Methods & Clinical Development |
| ISSN: | 2329-0501 |
| Popis: | The absence of fucose on asparagine-297 of the human immunoglobulin G (IgG) heavy chain has been shown to enhance antibody-dependent cellular cytotoxicity (ADCC) activity by 10- to 100-fold compared to fucosylated antibody. Our lab is studying the use of adeno-associated virus (AAV) as a vector for the delivery of HIV-specific antibodies for therapeutic purposes. Since the antibody is produced by vector-transduced cells in vivo, current techniques of glycoengineering cannot be utilized. In order to achieve similar enhancement of ADCC with AAV-delivered antibodies, short hairpin RNAs (shRNAs) that target fucosyltransferase-8 (FUT8), were designed, tested, and cloned into AAV vectors used to deliver HIV-specific broadly neutralizing antibodies (bNAbs). Antibodies produced by our glycoengineered-AAV (GE-AAV) vectors were analyzed for fucose content and ADCC. GE-AAV constructs were able to achieve over 80% knockdown of FUT8. Results were confirmed by lectin western blot for α1-6 fucose, which revealed almost a complete absence of fucose on GE-AAV-produced antibodies. GE-AAV-produced antibodies revealed >10-fold enhancement of ADCC, while showing identical neutralization and gp140 trimer binding compared to their fucosylated counterparts. ADCC was enhanced 40- to 60-fold when combined with key Fc mutations known to enhance binding to FcγRIIIA. Our findings define a powerful approach for supercharging AAV-delivered anti-HIV antibodies. Graphical Abstract Termini et al. describe the creation and validation of a novel glycoengineered-AAV vector for the delivery of anti-HIV antibodies. These AAV are capable of knocking down fucosyltransferase-8 following transduction and, when combined with antibody-Fc mutations, produce broadly neutralizing antibodies with 40- to 60-fold higher ADCC. |
| Databáze: | OpenAIRE |
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