Parallel in Vivo and in Vitro Selection Using Phage Display Identifies Protease-dependent Tumor-targeting Peptides

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...

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Bibliographic Details
Published in:The Journal of biological chemistry 2010-07, Vol.285 (29), p.22532-22541
Main Authors: Whitney, Mike, Crisp, Jessica L., Olson, Emilia S., Aguilera, Todd A., Gross, Larry A., Ellies, Lesley G., Tsien, Roger Y.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Bacteriophage
Breast Cancer
Cancer Therapy
Drug Delivery Systems - methods
Enzymology
Extracellular Matrix Proteins
Glycobiology and Extracellular Matrices
Matrix Metalloproteinase
Mice
Molecular Sequence Data
Neoplasms - metabolism
Neutrophil
Peptidases
Peptide Hydrolases - metabolism
Peptide Library
Peptides - chemistry
Peptides - pharmacology
Plasmin
Protease Inhibitor
Serine Protease
Tissue Extracts
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Summary: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.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M110.138297