Glycoengineering of AAV-delivered monoclonal antibodies yields increased ADCC activity

James M. Termini, José M. Martinez-Navio, Guangping Gao, Sebastian P. Fuchs, Ronald C. Desrosiers

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)204-217
Number of pages14
JournalMolecular Therapy - Methods and Clinical Development
Volume20
DOIs
StatePublished - Mar 12 2021

Keywords

  • adeno-associated virus
  • antibody-dependent cellular cytotoxicity
  • broadly neutralizing antibodies
  • fucosyltransferase 8
  • gene therapy
  • glycoengineering
  • immunotherapy
  • viral reservoir

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Fingerprint

Dive into the research topics of 'Glycoengineering of AAV-delivered monoclonal antibodies yields increased ADCC activity'. Together they form a unique fingerprint.

Cite this