Thermoresponsive Bacteriophage Nanocarrier as a Gene Delivery Vector Targeted to the Gastrointestinal Tract

Abstract

The use of the gastrointestinal tract as a site for the local delivery of DNA is an exciting prospect. In order to obtain an effective vector capable of delivering a gene of interest to target cells to achieve sufficient and sustained transgene expression, with minimal toxicity, we developed a new generation of filamentous bacteriophage. This particular bacteriophage was genetically engineered to display an arginine-glycine-aspartic acid (RGD) motif (an integrin-binding peptide) on the major coat protein pVIII and carry a mammalian DNA cassette. One unanticipated observation is the thermoresponsive behavior of engineered bacteriophage. This finding has led us to simplify the isolation method to purify bacteriophage particles from cell culture supernatant by low-temperature precipitation. Our results showed that, in contrast to non-surface modified, the RGD-modified bacteriophage was successfully used to deliver a transgene to mammalian cells. Our in vitro model of the human intestinal follicle-associated epithelium also demonstrated that bacteriophage particles were stable in simulated gastrointestinal fluids and able to cross the human intestinal barrier. In addition, we confirmed an adjuvant property of the engineered bacteriophage to induce nitric oxide production by macrophages. In conclusion, our study demonstrated the possibility of using bacteriophage for gene transfer in the gastrointestinal tract.

Arming Filamentous Bacteriophage, a Nature-Made Nanoparticle, for New Vaccine and Immunotherapeutic Strategies

“…Filamentous bacteriophages engineered for targeting of different tumors or tissue were used not only for the delivery of immunogenic epitopes as a fusion with the coat proteins but were also engineered for the targeted nanodelivery of therapeutic agents, drugs [69], fluorescent dyes for imaging and diagnostic [70], DNA cassettes [71], and siRNA [72].…”

“…Compared to other bacteriophages, Ff phages are extremely resistant, even in harsh environments as the gastrointestinal tract and can survive to a wide range of pH above 3 [110], and are resistant to enzymatic degradation with hydrolytic enzymes [71].…”

Investigation of intestinal transportation of peptide‐displaying bacteriophage particles using phage display method

“…The broad application of phage for delivery purposes have been also documented by several studies. For instance, the phage particles were used as nanocarriers in drug delivery, 43–48 gene delivery, 49–53 and antigen delivery system in vaccination 54,55 …”

The Presence of Bacteriophages in the Human Body: Good, Bad or Neutral?

“…Lehti et al (2017) described that phages could be internalized by eukaryotic cells by binding to moieties that resemble bacterial phage receptors [ 42 ]. The presence of an integrin-binding motif in genetically engineered filamentous phage was shown by Namdee et al (2018) [ 43 ]. Nguyen et al (2017) showed phage internalization by endocytosis [ 44 ] and that they were transferred through the Golgi apparatus [ 45 ].…”