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Published on: April 27, 2018
Metastatic prostate cancer cell-specific phage-like particles as a targeted gene-delivery system
Olusegun A Fagbohun1, Robert A Kazmierczak, Valery A Petrenko
1Cancer Research Center, Columbia, MO 65201, USA. EisenstarkA@missouri.edu.
Background:
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. 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.
Methods:
To 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.
Results:
Phage 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.
Conclusion:
Successful 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.
Insights
Researchers developed novel phage-like particles targeting prostate cancer cells. These particles, expressing emerald green fluorescent protein, demonstrate potential for targeted gene delivery in prostate cancer therapy.
Area of Science:
- Biotechnology
- Nanomedicine
- Oncology
Background:
- Effective tumor treatment requires targeted drug delivery via nanomedicines.
- Phage libraries offer a method for identifying tumor-specific protein ligands.
- Previous studies utilized phage proteins for targeted delivery to breast, prostate, and glioma cells.
Purpose of the Study:
- To identify and characterize novel phage clones specific for metastatic prostate cancer cells (PC-3M).
- To develop phage-like particles for targeted gene delivery in prostate cancer therapy.
Main Methods:
- Screening of a large landscape phage library (f8/8, 2 × 10^9 clones) against PC-3M cells.
- Characterization of phage-PC-3M cell interactions using phage capture assay, immunofluorescence, and electron microscopy.
- Conversion of a selected prostate cancer-specific phage into phage-like particles expressing emerald green fluorescent protein (EGFP).
Main Results:
- Phage clone EPTHSWAT exhibited high selectivity for PC-3M cells and was internalized by them.
- Phage-like particles harboring EGFP were successfully generated from the selected phage.
- EGFP expression was confirmed in PC-3M cells treated with these phage-like particles.
Conclusions:
- Phage-like particles targeting PC-3M cells show promise for therapeutic gene delivery.
- The successful development of EGFP-expressing phage-like particles validates their potential in prostate cancer treatment.
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