Engineered M13 Bacteriophage Nanocarriers for Intracellular Delivery of Exogenous Proteins to Human Prostate Cancer Cells

Abstract

The size, well-defined structure, and relatively high folding energies of most proteins allow them to recognize disease-relevant receptors that present a challenge to small molecule reagents. While multiple challenges must be overcome in order to fully exploit the use of protein reagents in basic research and medicine, perhaps the greatest challenge is their intracellular delivery to a particular diseased cell. Here, we describe the genetic and enzymatic manipulation of prostate cancer cell-penetrating M13 bacteriophage to generate nanocarriers for the intracellular delivery of functional exogenous proteins to a human prostate cancer cell line.

Bacteriophages and phage-inspired nanocarriers for targeted delivery of therapeutic cargos

“…Furthermore, Deporter at al. [55], used genetic manipulation of M13 phage to express a cell-penetrating peptide on the pIII protein and a biotin acceptor peptide (BAP) on the pIX protein. The BAP provided a biotin binding site in which a biotinylated exogenous therapeutic cargo could be attached, and taken up into prostate cancer cells (Figure 2).…”

“…The use of polymeric nanoparticles to deliver various therapeutic cargos has limitations with regard to the need for multiple synthetic and purification steps, and difficulties in designing the precise size, shape, composition, and rigidity of the carrier[55] Genetically-engineered nanoparticles may be solutions for these limitations with a well-defined set of external ligands for better targeting. Furthermore, delivery of multiple protein cargos can be achieved by these phage-based nanocarriers [55].…”

“…Furthermore, delivery of multiple protein cargos can be achieved by these phage-based nanocarriers [55]. For example a study engineered M13 phage to display a prostate cancer cell-penetrating peptide (Ypep2) in pIII and a biotin-acceptor peptide on pIX and horseradish peroxidase [HRP] as a reported molecule.…”

“…B) Further modification of BAP by biotin-streptavidin provides a binding site for conjugation of exogenous cargo to be transferred into the targeted cells. Reprinted with permission from[55]. Copyright (2016) American Chemical Society.…”

“…(B) Quantitative demonstration of image (A) results (SAV-HRP: streptavidin- horseradish peroxidase) (Reprinted from ref. [55] with permission from American Chemical Society).…”