In vitro selection of a peptide with high selectivity for cardiomyocytes in vivo.
One approach to targeted therapies for cardiovascular disease relies on isolating ligands that enhance the tissue-specific uptake of genes or drugs by heart cells. To obtain heart-targeting ligands, phage display biopanning was used to isolate a 20-mer peptide that binds to isolated primary cardiomyocytes. The isolated phage, PCM.1, displays the peptide WLSEAGPVVTVRALRGTGSW, and binds these cells 180 times better than a control phage from the library. Furthermore, phage displaying this peptide preferentially bind to cardiomyocytes when compared with a panel of other cell types. A BLAST search revealed that this peptide contains a 12 amino acid segment with sequence identity to a peptide in tenascin-X, an extracellular matrix protein. Synthetic peptides containing the complete 20-mer or a 12-mer tenascin peptide partially blocked phage binding to the cardiomyocytes. We developed a quantitative real-time PCR assay to assess uptake of this phage by tissues in vivo. Using this assay, preferential localization of the PCM.1 phage in heart was observed compared to the uptake of this phage by other tissues or other phage by heart. Furthermore, PCM.1 phage was associated with cardiomyocytes isolated from mice treated with a phage in vivo. These results demonstrate the utility of biopanning on isolated cells for identifying specific binding peptides that can target a tissue in vivo.
Department of Internal Medicine, Center for Biomedical Inventions, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas 75390-9185, USA.
Michael J
MJ
McGuire
Kausar N
KN
Samli
Stephen Albert
SA
Johnston
Kathlynn C
KC
Brown
D000595:Amino Acid Sequence; D000818:Animals; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism; D002318:Cardiovascular Diseases / Q000188:drug therapy; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D051379:Mice; D008807:Mice, Inbred BALB C; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D019151:Peptide Library; D010455:Peptides / Q000235:genetics* / Q000378:metabolism* / Q000627:therapeutic use; D016133:Polymerase Chain Reaction / Q000379:methods; D011485:Protein Binding; D019063:Tenascin / Q000235:genetics
D019151:Peptide Library; D010455:Peptides; D019063:Tenascin; C081865:tenascin X