The transduction of Coxsackie and Adenovirus Receptor-negative cells and protection against neutralizing antibodies by HPMA-co-oligolysine copolymer-coated adenovirus

Biomaterials. 2011 Dec;32(35):9536-45. doi: 10.1016/j.biomaterials.2011.08.069. Epub 2011 Sep 28.

Abstract

Adenoviral (AdV) gene vectors offer efficient nucleic acid transfer into both dividing and non-dividing cells. However issues such as vector immunogenicity, toxicity and restricted transduction to receptor-expressing cells have prevented broad clinical translation of these constructs. To address this issue, engineered AdV have been prepared by both genetic and chemical manipulation. In this work, a polymer-coated Ad5 formulation is optimized by evaluating a series of N-(2-hydroxypropyl) methacrylamide (HPMA)-co-oligolysine copolymers synthesized by living polymerization techniques. This synthesis approach was used to generate highly controlled and well-defined polymers with varying peptide length (K(5), K(10) and K(15)), polymer molecular weight, and degradability to coat the viral capsid. The optimal formulation was not affected by the presence of serum during transduction and significantly increased Ad5 transduction of several cell types that lack the Coxsackie and Adenovirus Receptor (CAR) by up to 6-fold compared to unmodified AdV. Polymer-coated Ad5 also retained high transduction capability in the presence of Ad5 neutralizing antibodies. The critical role of heparan sulfate proteoglycans (HSPGs) in mediating cell binding and internalization of polymer-coated AdV was also demonstrated by evaluating transduction in HSPG-defective recombinant CHO cells. The formulations developed here are attractive vectors for ex vivo gene transfer in applications such as cell therapy. In addition, this platform for adenoviral modification allows for facile introduction of alternative targeting ligands.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acrylamides / chemistry*
  • Adenoviridae / drug effects
  • Adenoviridae / metabolism*
  • Animals
  • Antibodies, Neutralizing / pharmacology*
  • CHO Cells
  • Coxsackie and Adenovirus Receptor-Like Membrane Protein
  • Cricetinae
  • Cricetulus
  • Cytoprotection / drug effects*
  • HeLa Cells
  • Heparan Sulfate Proteoglycans / genetics
  • Humans
  • Mice
  • Microscopy, Electron, Transmission
  • Mutation / genetics
  • Polylysine / analogs & derivatives*
  • Polylysine / chemistry
  • Polymerization / drug effects
  • Receptors, Virus / metabolism*
  • Transduction, Genetic / methods*
  • Virus Internalization / drug effects

Substances

  • Acrylamides
  • Antibodies, Neutralizing
  • CLMP protein, human
  • CLMP protein, mouse
  • Coxsackie and Adenovirus Receptor-Like Membrane Protein
  • Heparan Sulfate Proteoglycans
  • Receptors, Virus
  • Polylysine
  • N-(2-hydroxypropyl)methacrylamide