Parallel in vivo and in vitro selection using phage display identifies protease-dependent tumor-targeting peptides

Mike Whitney, Jessica L. Crisp, Emilia S. Olson, Todd A. Aguilera, Larry A. Gross, Lesley G. Ellies, Roger Y. Tsien

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

We recently developed activatable cell-penetrating peptides (ACPPs) that target contrast agents to in vivo sites of matrix metalloproteinase activity, such as tumors. Here we use parallel in vivo and in vitro selection with phage display to identify novel tumor-homing ACPPs with no bias for primary sequence or target protease. Specifically, phage displaying a library of ACPPs were either injected into tumor-bearing mice, followed by isolation of cleaved phage from dissected tumor, or isolated based on selective cleavage by extracts of tumor versus normal tissue. Selected sequences were synthesized as fluorescently labeled peptides, and tumor-specific cleavage was confirmed by digestion with tissue extracts. The most efficiently cleaved peptide contained the substrate sequence RLQLKL and labeled tumors and metastases from several cancer models with up to 5-fold contrast. This uniquely identified ACPP was not cleaved by matrix metalloproteinases or various coagulation factors but was efficiently cleaved by plasmin and elastases, both of which have been shown to be aberrantly overexpressed in tumors. The identification of an ACPP that targets tumor expressed proteases without rational design highlights the value of unbiased selection schemes for the development of potential therapeutic agents.

Original languageEnglish (US)
Pages (from-to)22532-22541
Number of pages10
JournalJournal of Biological Chemistry
Volume285
Issue number29
DOIs
StatePublished - Jul 16 2010

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Bacteriophages
Tumors
Peptide Hydrolases
Cell-Penetrating Peptides
Display devices
Peptides
Neoplasms
Matrix Metalloproteinases
Bearings (structural)
In Vitro Techniques
Tissue Extracts
Blood Coagulation Factors
Pancreatic Elastase
Fibrinolysin
Contrast Media
Libraries
Digestion
Tissue
Neoplasm Metastasis

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Parallel in vivo and in vitro selection using phage display identifies protease-dependent tumor-targeting peptides. / Whitney, Mike; Crisp, Jessica L.; Olson, Emilia S.; Aguilera, Todd A.; Gross, Larry A.; Ellies, Lesley G.; Tsien, Roger Y.

In: Journal of Biological Chemistry, Vol. 285, No. 29, 16.07.2010, p. 22532-22541.

Research output: Contribution to journalArticle

Whitney, Mike ; Crisp, Jessica L. ; Olson, Emilia S. ; Aguilera, Todd A. ; Gross, Larry A. ; Ellies, Lesley G. ; Tsien, Roger Y. / Parallel in vivo and in vitro selection using phage display identifies protease-dependent tumor-targeting peptides. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 29. pp. 22532-22541.
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