Abstract
Cell-penetrating peptides (CPPs), also known as protein transduction domains, were first identified 25 years ago. They are small, ~6–30 amino acid long, synthetic, or naturally occurring peptides, able to carry a variety of cargoes across the cellular membranes in an intact, functional form. These cargoes can range from other small peptides, full-length proteins, nucleic acids including RNA and DNA, nanoparticles, and viral particles as well as radioisotopes and other fluorescent probes for imaging purposes. However, this ability to enter all cell types indiscriminately, and even cross the blood–brain barrier, hinders their development into viable vectors. Hence, researchers have adopted various strategies ranging from pH activatable cargoes to using phage display to identify tissue-specific CPPs. Use of this phage display strategy has led to an ever-expanding number of tissue-specific CPPs. Using phage display, we identified a 12-amino acid, non-naturally occurring peptide that targets the heart with peak uptake at 15 min after a peripheral intravenous injection, that we termed Cardiac Targeting Peptide (CTP). In this chapter, we use CTP as an example to describe techniques for validation of cell-specific transduction as well as provide details on a technology to identify binding partner(s) for these ever-increasing plethora of tissue-specific peptides. Given the myriad cargoes CTP can deliver, as well as rapid uptake after an intravenous injection, it can be applied to deliver radioisotopes, miRNA, siRNA, peptides, and proteins of therapeutic potential for acute cardiac conditions like myocardial infarction, where the window of opportunity for salvaging at-risk myocardium is limited to 6 hrs.
Key words
- Cardiac targeting peptide
- Protein transduction domains
- Cell-penetrating peptides
- Phage display
- TriCEPS
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Acknowledgments
M.Z. and K.S.F. are supported by American Heart Association Scientist Development Award 17SDG33411180, and by a grant awarded under the Pitt Innovation Challenge (PinCh) through the Clinical and Translational Science Institute of the University of Pittsburgh, through National Institutes of Health, UL1TR001857.
Disclosures: M.Z. along with Paul D. Robbins (Professor, University of Minnesota, Minnesota, MN, USA) hold a patent on the use of cardiac targeting peptide as a cardiac vector (Cardiac-specific protein targeting domain, U.S. Patent Serial No. 9,249,184). M.Z. also serves as Chief Scientific Officer and on the Board of Directors of the startup Vivasc Therapeutics Inc., and holds substantial equity in it. M.P. is the Chief Scientific Officer of Dualsystems Biotech AG that holds an exclusive license for the LRC technology as covered in patent application WO2012/104051.
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Feldman, K.S., Pavlou, M.P., Zahid, M. (2021). Cardiac Targeting Peptide : From Identification to Validation to Mechanism of Transduction. In: Narayanan, K. (eds) Bio-Carrier Vectors. Methods in Molecular Biology, vol 2211. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0943-9_8
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DOI: https://doi.org/10.1007/978-1-0716-0943-9_8
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