Adenoviral vectors coated with PAMAM dendrimer conjugates allow CAR independent virus uptake and targeting to the EGF receptor.

Abstract

Adenovirus type 5 (Ad) is an efficient gene vector with high gene transduction potential, but its efficiency depends on its native cell receptors coxsackie- and adenovirus receptor (CAR) for cell attachment and α(v)β(3/5) integrins for internalization. To enable transduction of CAR negative cancer cell lines, we have coated the negatively charged Ad by noncovalent charge interaction with cationic PAMAM (polyamidoamine) dendrimers. The specificity for tumor cell infection was increased by targeting the coated Ad to the epidermal growth factor receptor using the peptide ligand GE11, which was coupled to the PAMAM dendrimer via a 2 kDa PEG spacer. Particles were examined by measuring surface charge and size, the degree of coating was determined by transmission electron microscopy. The net positive charge of PAMAM coated Ad enhanced cellular binding and uptake leading to increased transduction efficiency, especially in low to medium CAR expressing cancer cell lines using enhanced green fluorescent protein or luciferase as transgene. While PAMAM coated Ad allowed for efficient internalization, coating with linear polyethylenimine induced excessive particle aggregation, elevated cellular toxicity and lowered transduction efficiency. PAMAM coating of Ad enabled successful transduction of cells in vitro even in the presence of neutralizing antibodies. Taken together, this study clearly proves noncovalent, charge-based coating of Ad vectors with ligand-equipped dendrimers as a viable strategy for efficient transduction of cells otherwise refractory to Ad infection.

• Authors: Alexandra Vetter, Kulpreet S Virdi, Sigrid Espenlaub, Wolfgang Rödl, Ernst Wagner, Per S Holm, Christina Scheu, Florian Kreppel, Christine Spitzweg, Manfred Ogris
• Journal: Molecular pharmaceutics (Mol Pharm) Vol. 10 Issue 2 Pg. 606-18 (Feb 04 2013) ISSN: 1543-8392 [Electronic] United States
• PMID: 23281933 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Coxsackie and Adenovirus Receptor-Like Membrane Protein
• Dendrimers
• PAMAM Starburst
• ErbB Receptors

Topics

• Adenoviridae
• Cell Line, Tumor
• Cell Survival (genetics, physiology)
• Coxsackie and Adenovirus Receptor-Like Membrane Protein (genetics, metabolism)
• Dendrimers (chemistry)
• ErbB Receptors (genetics, metabolism)
• Flow Cytometry
• Genetic Vectors (chemistry)
• Humans
• Microscopy, Electron, Transmission