title: Identification of a cardiac specific protein transduction domain by in vivo biopanning using a M13 phage peptide display library in mice.
abstract: BACKGROUND
  A peptide able to transduce cardiac tissue specifically, delivering cargoes to the heart, would be of significant therapeutic potential for delivery of small molecules, proteins and nucleic acids. In order to identify peptide(s) able to transduce heart tissue, biopanning was performed in cell culture and in vivo with a M13 phage peptide display library.
  
  
  RESULTS
  A cardiomyoblast cell line, H9C2, was incubated with a M13 phage 12 amino acid peptide display library. Internalized phage was recovered, amplified and then subjected to a total of three rounds of in vivo biopanning where infectious phage was isolated from cardiac tissue following intravenous injection. After the third round, 60% of sequenced plaques carried the peptide sequence APWHLSSQYSRT, termed cardiac targeting peptide (CTP). We demonstrate that CTP was able to transduce cardiomyocytes functionally in culture in a concentration and cell-type dependent manner. Mice injected with CTP showed significant transduction of heart tissue with minimal uptake by lung and kidney capillaries, and no uptake in liver, skeletal muscle, spleen or brain. The level of heart transduction by CTP also was greater than with a cationic transduction domain.
  
  
  CONCLUSIONS
  Biopanning using a peptide phage display library identified a peptide able to transduce heart tissue in vivo efficiently and specifically. CTP could be used to deliver therapeutic peptides, proteins and nucleic acid specifically to the heart.
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Maliha
authors.initials: M
authors.lastname: Zahid
authors.forename: Brett E
authors.initials: BE
authors.lastname: Phillips
authors.forename: Sean M
authors.initials: SM
authors.lastname: Albers
authors.forename: Nick
authors.initials: N
authors.lastname: Giannoukakis
authors.forename: Simon C
authors.initials: SC
authors.lastname: Watkins
authors.forename: Paul D
authors.initials: PD
authors.lastname: Robbins
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017104:Bacteriophage M13; D001692:Biological Transport; D018929:Cell Culture Techniques; D002460:Cell Line; D005260:Female; D006801:Humans; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D032386:Myoblasts, Cardiac / Q000166:cytology / Q000378:metabolism; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D009928:Organ Specificity; D019151:Peptide Library; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D000097782:NF-kappaB-Inducing Kinase
chemical_list: D026901:Membrane Transport Proteins; D016328:NF-kappa B; D009842:Oligopeptides; D019151:Peptide Library; D017346:Protein Serine-Threonine Kinases; D000097782:NF-kappaB-Inducing Kinase
references.citation: Nature. 1996 Mar 28;380(6572):364-6
references.pmid: 8598934
references.citation: Bioorg Med Chem Lett. 2008 Jul 15;18(14):4011-4
references.pmid: 18565751
references.citation: Virology. 2002 Feb 1;293(1):182-91
references.pmid: 11853411
references.citation: Biomaterials. 2009 Mar;30(7):1287-98
references.pmid: 19095299
references.citation: J Am Coll Cardiol. 2004 Aug 18;44(4):783-9
references.pmid: 15312859
references.citation: Science. 1998 Jan 16;279(5349):377-80
references.pmid: 9430587
references.citation: Am J Pathol. 2005 Feb;166(2):625-36
references.pmid: 15681844
references.citation: Mol Imaging Biol. 2006 Jul-Aug;8(4):201-7
references.pmid: 16791746
references.citation: J Am Coll Cardiol. 2000 Dec;36(7):2056-63
references.pmid: 11127441
references.citation: J Mol Biol. 2004 Sep 3;342(1):171-82
references.pmid: 15313615
references.citation: Circulation. 2005 Sep 13;112(11):1601-11
references.pmid: 16144998
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 17;101(33):12282-7
references.pmid: 15302924
references.citation: Nat Med. 2004 Jun;10(6):625-32
references.pmid: 15133506
references.citation: Am J Physiol Heart Circ Physiol. 2006 Sep;291(3):H1337-44
references.pmid: 16632551
references.citation: Mol Ther. 2003 Aug;8(2):295-305
references.pmid: 12907152