Laryngeal muscle surface receptors identified using random phage library.
OBJECTIVE
The ultimate goal of this study is to improve the efficiency of gene transfer in mammalian muscle by developing targeted adenoviral vectors. Altering the tropism of viral vectors to recognize tissue specific antigens is one method to achieve this goal. This approach requires identification of cell-surface receptors and the insertion of target peptide sequences into the adenoviral fiber protein. In this study, phage biopanning was performed on cultured rat skeletal and laryngeal muscle to identify cell-surface receptors.


METHODS
In vitro cell culture and in vivo animal model.


METHODS
M-13 Phage biopanning was used for muscle cell-surface receptor analysis on cultured rat skeletal and laryngeal muscle. Nonbinding and binding phage to cultured skeletal and laryngeal muscle were screened for muscle specific surface peptides. In vivo studies were then performed using muscle specific phage.


RESULTS
Skeletal muscle specific binding by the YASTNPM phage was observed by in vivo immunostaining. Phage titering demonstrated a 10(9)-fold increase in skeletal muscle binding compared with nontarget tissue. A peptide sequence (NPSQVKH) specific for laryngeal muscle yielded a 10(7)-fold increase in laryngeal muscle phage titer compared with nontarget tissue.


CONCLUSIONS
These results identify muscle cell-surface receptors that may be used as potential targets for genetic modification of adenovirus tropism. Moreover, phage specificity for skeletal and laryngeal muscle indicates specific muscle groups may be targeted.
Flint
Paul W
PW
Department of Otolaryngology-Head and Neck Surgery Johns Hopkins University, Baltimore, Maryland, USA. pflint@jhmi.edu
Li
Zhao Bo
ZB
Lehar
Mohamed
M
Saito
Koichiro
K
Pai
Sara I
SI
D000256:Adenoviridae; D000818:Animals; D001435:Bacteriophages; D002460:Cell Line; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000627:therapeutic use; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007821:Laryngeal Muscles / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism / Q000821:virology; D019151:Peptide Library; D051381:Rats; D017207:Rats, Sprague-Dawley; D011956:Receptors, Cell Surface / Q000378:metabolism*
D019151:Peptide Library; D011956:Receptors, Cell Surface