Development of a Novel Cytokine Vehicle Using Filamentous Phage Display for Colorectal Cancer Treatment

Abstract

Due to its highly immunogenic nature and the great engineerability, filamentous phage has shown promising antitumor activities in preclinical studies. Previous designs of antitumor phage mainly focused on tumor targeting using a cancerspecific moiety displayed on the minor capsid protein, pIII. In this work, we developed a new therapeutic platform of filamentous phage, in which the major capsid protein pVIII was utilized for displaying an antitumor cytokine. We showcased that a 16.1-kD cytokine GM-CSF could be efficiently presented on the M13 phage particle using the 8 + 8 type display system through a highly tolerable pVIII variant P8(1a). We verified that the GM-CSF phage was a potent activator for STAT5 signaling in murine macrophage. The GM-CSF phage significantly reduced the tumor size by more than 50% as compared to the unmodified phage in a murine colorectal cancer model. Immunological profiling of the tumor-infiltrating leukocytes revealed that an increase of CD4+ lymphocytes in the GM-CSF phage treatment group. Furthermore, the combined therapy of the GM-CSF phage and radiation greatly improved the therapeutic potency with a 100% survival rate and a 25% complete remission rate. We observed that the IFN-γ expression was dramatically up-regulated by the combined therapy in multiple types of tumor-infiltrating immune cells. Overall, we created a novel vehicle for cytokine therapy using the pVIII filamentous phage display. This new platform can be multiplexed with other phage engineering approaches, such as displaying targeting ligands on pIII or encapsulating therapeutic genes inside phage capsids, to create multifunctional nanoparticles for cancer therapy.

The Effect of Zero-Valent Iron Nanoparticles (nZVI) on Bacteriophages

“…Inoviridae , comprising, among others, M13, f1, and fd is the most broadly investigated order as nanocarriers [ 53 ]. Filamentous bacteriophages find wide applications as antitumor agents utilizing phage display techniques [ 54 ]. They are also readily genetically engineered using well-established cloning methods and are highly immunogenic [ 53 ].…”

A genetically engineered phage-based nanomaterial for detecting bacteria with magnetic resonance imaging

“…In addition, theoretical considerations indicate that phage relaxivity should increase at lower fields (due to slowing down of molecular tumbling), which could also contribute to enhancing contrast at clinically-relevant field strengths (e.g., 1.5 Tesla), albeit with a tradeoff in overall signal intensity. In summary, as a phage-based molecular probe for visualizing targeted bacterial strains with MRI, we envision a synergy in the development of DP1-phage technology with rapidly evolving advances in emerging biotherapeutic platforms involving the use of live microorganisms as well as bacteriophage formulations for noninvasive diagnostics and treatment of diseases like cancer [46][47][48][49][50][51][52] , inflammatory conditions 53,54 , metabolic disorders 55,56 , and antibioticresistant infections [57][58][59][60][61] .…”

Phage display for the detection, analysis, disinfection, and prevention of Staphylococcus aureus

“…The applications for phage display technology are numerous. 31,38,39 Phage display is commonly used in protein engineering. For this reason, the technique can be incredibly useful for drug discovery and development.…”

“…The applications for phage display technology are numerous 31,38,39 . Phage display is commonly used in protein engineering.…”

“…For this reason, the technique can be incredibly useful for drug discovery and development. A common application of this technique has been employed to develop new treatments for cancer either through identifying tumor antigens or by expressing specific epitopes that could elicit strong immunological responses 38,40 …”