The zebrafish embryo as a tool for screening and characterizing pleurocidin host-defense peptides as anti-cancer agents

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DOIResolve DOI: http://doi.org/10.1242/dmm.007310
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TypeArticle
Journal titleDisease Models & Mechanisms
Volume4
Issue5
Pages622633; # of pages: 12
Subjectcytotoxic agent; nrc 01; nrc 02; nrc 03; nrc 03b; nrc 03d; nrc 04; nrc 05; nrc 06; nrc 07; nrc 08; nrc 09; nrc 10; nrc 11; nrc 12; nrc 123; nrc 124; nrc 125; nrc 126; nrc 127; nrc 128; nrc 13; nrc 14; nrc 15; nrc 16; nrc 17; nrc 18; nrc 19; nrc 20; unclassified drug; unindexed drug
AbstractThe emergence of multidrug-resistant cancers and the lack of targeted therapies for many cancers underscore an unmet need for new therapeutics with novel modes of action towards cancer cells. Host-defense peptides often exhibit selective cytotoxicity towards cancer cells and show potential as anti-cancer therapeutics. Here, we screen 26 naturally occurring variants of the peptide pleurocidin for cytotoxic and anti-cancer activities, and investigate the underlying mechanism of action. Cytotoxicities were assessed in vitro using cell-based assays and in vivo using zebrafish embryos. Morphological changes were assessed by both transmission and scanning electron microscopy, and functional assays were performed on zebrafish embryos to investigate the mechanism of cell death. A total of 14 peptides were virtually inactive against HL60 human leukemia cells, whereas 12 caused >50% death at ≤32 &μυ;g/ml. Morphological changes characteristic of oncosis were evident by electron microscopy after only 1 minute of treatment with 32 &μυ;g/ml of variant NRC-03. Only two peptides were hemolytic. Four peptides showed no toxicity towards zebrafish embryos at the highest concentration tested (25 &μυ;M; ~64 &μυ;g/ml) and one peptide was highly toxic, killing 4-hour-post-fertilization (hpf) embryos immediately after exposure to 1 &μυ;M peptide. Four other peptides killed embryos after 24 hours of exposure at 1 &μυ;M. Most peptides caused mortality at one or more developmental stages only after continuous exposure (24 hours) with higher lethal doses (≥5 &μυ;M). Pleurocidin NRC-03 bound to embryos and induced the release of superoxide, caused an increase in the number of TUNEL-positive nuclei, and caused membrane damage and the loss of embryonic epithelial integrity, marked by the exclusion of cells from the outer epithelium and the appearance of F-actin within the circumferential cells of the repair site. Our results indicate that specific pleurocidin variants are attractive cancer-selective agents that selectively induce cell death in target cells but leave non-target cells such as erythrocytes and non-transformed cells unaffected.
Publication date
LanguageEnglish
AffiliationNRC Institute for Marine Biosciences; NRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedYes
NPARC number19709132
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Record identifierbdb3dec2-34e1-4030-825b-d5b57ee07f75
Record created2012-04-02
Record modified2016-05-09
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