Abstract
BackgroundOne of the cardinal requirements for effective therapeutic management of tumors is the selective delivery of cancer drugs to the right site by ligand-decorated nanomedicines. Screening of 2 × 109 clone landscape phage library provides a reliable avenue for generating protein ligands specific for tumor cells. It was shown that selective phage proteins derived from landscape phage libraries against breast and prostate cancer cells are able to navigate drug or siRNA loaded liposomes to corresponding cancer cells with minimal toxicity to non-neoplastic cells. In an alternative platform, glioma cell-specific phage proteins were used for assembling in vivo cancer-specific phage-like particles, named ‘phagemid infective particles’ as targeted gene-delivery vehicles.MethodsTo extend the panel of anticancer cell phages, we have screened a 2 × 109 clone landscape phage library f8/8 to select phage clones specific for metastatic prostate cancer cell PC-3M. The phage clones were characterized for their selective interaction with PC-3M cells using phage capture assay, immunofluorescence microscopy and electron microscopy. A prostate cancer selective phage was converted to phage-like particles harboring emerald green fluorescent protein.ResultsPhage clone EPTHSWAT (designated by the sequence of inserted peptide) was found to be most selective for PC-3M cells and was observed to internalize PC-3M cells as revealed by immunofluorescence microscopy and electron microscopy. Conversion of this phage to phage-like particles harboring emerald green fluorescent protein and the expression of emerald green fluorescent protein in the phage-like particles treated PC-3M cells showed potential of adoption of this phage-like particle in prostate cancer therapeutic gene delivery.ConclusionSuccessful employment of phage-like particles expressing emerald green fluorescent protein genes targeted to prostate cancer cells PC-3M confirms a prospect of their use for targeted delivery of therapeutic genes to cancer cells.
Highlights
One of the cardinal requirements for effective therapeutic management of tumors is the selective delivery of cancer drugs to the right site by ligand-decorated nanomedicines
Selection of prostate cancer cell-interacting phage clones and studying their selectivity towards the target cells To generate peptides specific for prostate cancer cells, landscape phage library with an octapeptide insert was screened against the metastatic prostate cancer cell line PC-3M involving two selection rounds. 18 clones were randomly picked and the structures of the displayed peptides were determined using PCR, sequencing and translation of the DNA sequences into amino acids using DNASTAR software (DNASTAR, Madison, WI) (Table 1)
Selectivity of the two abundant prostate cancerspecific phages EPTHSWAT and GDIVTSNS towards PC-3M cells were studied in a phage capture assay (Figure 1)
Summary
One of the cardinal requirements for effective therapeutic management of tumors is the selective delivery of cancer drugs to the right site by ligand-decorated nanomedicines. In another study, [14] developed phagemid particles with epidermal growth factors fused to pIII to transduce mammalian cells Another type of display, with fusion peptides displayed on all multiple copies of the major coat protein pVIII can be even more advantageous for cell transduction since the multivalent display results in more strong binding of the phage to cellular receptors due to avidity effect, which provokes receptor dimerization and clustering [3,15,16]. With fusion peptides displayed on all multiple copies of the major coat protein pVIII can be even more advantageous for cell transduction since the multivalent display results in more strong binding of the phage to cellular receptors due to avidity effect, which provokes receptor dimerization and clustering [3,15,16] Another type of tumor selective phagemid gene-delivery vehicles known as phagemid infective particle was designed by [17] using a phagemid system. Thereafter, the phage clone was converted into phagelike particles using the phagemid system and its PC-3M cell-transforming activity was observed using emerald green fluorescent protein marker
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