Identification of a cardiac specific protein transduction domain by in vivo biopanning using a M13 phage peptide display library in mice

PLoS One. 2010 Aug 17;5(8):e12252. doi: 10.1371/journal.pone.0012252.

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.

Methods and 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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacteriophage M13*
  • Biological Transport
  • Cell Culture Techniques
  • Cell Line
  • Female
  • Humans
  • Membrane Transport Proteins / chemistry
  • Membrane Transport Proteins / metabolism*
  • Mice
  • Microscopy, Confocal
  • Myoblasts, Cardiac / cytology
  • Myoblasts, Cardiac / metabolism
  • Myocardium / cytology
  • Myocardium / metabolism*
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / metabolism
  • NF-kappa B / metabolism
  • NF-kappaB-Inducing Kinase
  • Oligopeptides / chemistry
  • Oligopeptides / metabolism*
  • Organ Specificity
  • Peptide Library*
  • Protein Serine-Threonine Kinases / metabolism
  • Protein Structure, Tertiary
  • Signal Transduction

Substances

  • Membrane Transport Proteins
  • NF-kappa B
  • Oligopeptides
  • Peptide Library
  • Protein Serine-Threonine Kinases