title: Impact of capsid modifications by selected peptide ligands on recombinant adeno-associated virus serotype 2-mediated gene transduction.
abstract: Vectors based on adeno-associated virus serotype 2 (AAV2) belong to today's most promising and most frequently used viral vectors in human gene therapy. Like in many other vector systems, the broad but non-specific tropism limits their use for certain cell types or tissues. One approach to screen for transduction-improved vectors is the selection of random peptide libraries displayed directly on the AAV2 capsid. Although the AAV2 library system has been widely applied for the successful selection of improved gene therapy vectors, it remains unknown which steps of the transduction process are most affected and therefore critical for the selection of targeting peptides. Attachment to the cell surface is the first essential step of AAV-mediated gene transduction; however, our experiments challenge the conventional belief that enhanced gene transfer is equivalent to more efficient cell binding of recombinant AAV2 vectors. A comparison of the various steps of gene transfer by vectors carrying a wild-type AAV2 capsid or displaying two exemplary peptide ligands selected from AAV2 random libraries on different human tumour cell lines demonstrated strong alterations in cell binding, cellular uptake, as well as intracellular processing of these vectors. Combined, our results suggest that entry and post-entry events are decisive for the selection of the peptides NDVRSAN and GPQGKNS rather than their cell binding efficiency.
authors: German Cancer Research Center (DKFZ), Department of Tumorvirology, Im Neuenheimer Feld 242, 69120, Heidelberg, Germany.
authors: Matthias
authors: M
authors: Naumer
authors: German Cancer Research Center (DKFZ), Department of Tumorvirology, Im Neuenheimer Feld 242, 69120, Heidelberg, Germany.
authors: Ruth
authors: R
authors: Popa-Wagner
authors: German Cancer Research Center (DKFZ), Department of Tumorvirology, Im Neuenheimer Feld 242, 69120, Heidelberg, Germany.
authors: Jürgen A
authors: JA
authors: Kleinschmidt
mesh_terms: D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism*; D045744:Cell Line, Tumor; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D005909:Glioblastoma / Q000235:genetics / Q000378:metabolism / Q000821:virology; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D008545:Melanoma / Q000235:genetics / Q000378:metabolism / Q000821:virology; D019151:Peptide Library; D010455:Peptides / Q000235:genetics* / Q000378:metabolism*; D014161:Transduction, Genetic; D014779:Virus Replication / Q000235:genetics
chemical_list: D036022:Capsid Proteins; D008024:Ligands; D019151:Peptide Library; D010455:Peptides