Successful expansion but not complete restriction of tropism of adeno-associated virus by in vivo biopanning of random virus display peptide libraries
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Successful expansion but not complete restriction of tropism of adeno-associated virus by in vivo biopanning of random virus display peptide libraries. / Michelfelder, Stefan; Kohlschütter, Johannes; Skorupa, Alexandra; Pfennings, Sabrina; Müller, Oliver; Kleinschmidt, Jürgen A; Trepel, Martin.
in: PLOS ONE, Jahrgang 4, Nr. 4, 2009, S. e5122.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Successful expansion but not complete restriction of tropism of adeno-associated virus by in vivo biopanning of random virus display peptide libraries
AU - Michelfelder, Stefan
AU - Kohlschütter, Johannes
AU - Skorupa, Alexandra
AU - Pfennings, Sabrina
AU - Müller, Oliver
AU - Kleinschmidt, Jürgen A
AU - Trepel, Martin
PY - 2009
Y1 - 2009
N2 - Targeting viral vectors to certain tissues in vivo has been a major challenge in gene therapy. Cell type-directed vector capsids can be selected from random peptide libraries displayed on viral capsids in vitro but so far this system could not easily be translated to in vivo applications. Using a novel, PCR-based amplification protocol for peptide libraries displayed on adeno-associated virus (AAV), we selected vectors for optimized transduction of primary tumor cells in vitro. However, these vectors were not suitable for transduction of the same target cells under in vivo conditions. We therefore performed selections of AAV peptide libraries in vivo in living animals after intravenous administration using tumor and lung tissue as prototype targets. Analysis of peptide sequences of AAV clones after several rounds of selection yielded distinct sequence motifs for both tissues. The selected clones indeed conferred gene expression in the target tissue while gene expression was undetectable in animals injected with control vectors. However, all of the vectors selected for tumor transduction also transduced heart tissue and the vectors selected for lung transduction also transduced a number of other tissues, particularly and invariably the heart. This suggests that modification of the heparin binding motif by target-binding peptide insertion is necessary but not sufficient to achieve tissue-specific transgene expression. While the approach presented here does not yield vectors whose expression is confined to one target tissue, it is a useful tool for in vivo tissue transduction when expression in tissues other than the primary target is uncritical.
AB - Targeting viral vectors to certain tissues in vivo has been a major challenge in gene therapy. Cell type-directed vector capsids can be selected from random peptide libraries displayed on viral capsids in vitro but so far this system could not easily be translated to in vivo applications. Using a novel, PCR-based amplification protocol for peptide libraries displayed on adeno-associated virus (AAV), we selected vectors for optimized transduction of primary tumor cells in vitro. However, these vectors were not suitable for transduction of the same target cells under in vivo conditions. We therefore performed selections of AAV peptide libraries in vivo in living animals after intravenous administration using tumor and lung tissue as prototype targets. Analysis of peptide sequences of AAV clones after several rounds of selection yielded distinct sequence motifs for both tissues. The selected clones indeed conferred gene expression in the target tissue while gene expression was undetectable in animals injected with control vectors. However, all of the vectors selected for tumor transduction also transduced heart tissue and the vectors selected for lung transduction also transduced a number of other tissues, particularly and invariably the heart. This suggests that modification of the heparin binding motif by target-binding peptide insertion is necessary but not sufficient to achieve tissue-specific transgene expression. While the approach presented here does not yield vectors whose expression is confined to one target tissue, it is a useful tool for in vivo tissue transduction when expression in tissues other than the primary target is uncritical.
KW - Amino Acid Sequence
KW - Animals
KW - Breast Neoplasms
KW - Capsid
KW - Dependovirus
KW - Female
KW - Gene Transfer Techniques
KW - Genetic Therapy
KW - Genetic Vectors
KW - Humans
KW - Mice
KW - Mice, Transgenic
KW - Molecular Sequence Data
KW - Peptide Library
KW - Polymerase Chain Reaction
KW - Transduction, Genetic
KW - Tropism
KW - Tumor Cells, Cultured
KW - Evaluation Studies
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1371/journal.pone.0005122
DO - 10.1371/journal.pone.0005122
M3 - SCORING: Journal article
C2 - 19357785
VL - 4
SP - e5122
JO - PLOS ONE
JF - PLOS ONE
SN - 1932-6203
IS - 4
ER -