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Hybrid Nanomaterial Complexes for Advanced Phage-guided Gene Delivery.

Yata T, Lee KY, Dharakul T, Songsivilai S, Bismarck A, Mintz PJ, Hajitou A - Mol Ther Nucleic Acids (2014)

Bottom Line: We demonstrate that the phage complex with cationic polymers generates positively charged phage and large aggregates that show enhanced cell surface attachment, buffering capacity, and improved transgene expression while retaining cell type specificity.Moreover, phage/polymer complexes carrying a therapeutic gene achieve greater cancer cell killing than phage alone.This new class of hybrid nanomaterial platform can advance targeted gene delivery applications by bacteriophage.

View Article: PubMed Central - PubMed

Affiliation: Phage Therapy Group, Department of Medicine, Imperial College London, London, UK.

ABSTRACT
Developing nanomaterials that are effective, safe, and selective for gene transfer applications is challenging. Bacteriophages (phage), viruses that infect bacteria only, have shown promise for targeted gene transfer applications. Unfortunately, limited progress has been achieved in improving their potential to overcome mammalian cellular barriers. We hypothesized that chemical modification of the bacteriophage capsid could be applied to improve targeted gene delivery by phage vectors into mammalian cells. Here, we introduce a novel hybrid system consisting of two classes of nanomaterial systems, cationic polymers and M13 bacteriophage virus particles genetically engineered to display a tumor-targeting ligand and carry a transgene cassette. We demonstrate that the phage complex with cationic polymers generates positively charged phage and large aggregates that show enhanced cell surface attachment, buffering capacity, and improved transgene expression while retaining cell type specificity. Moreover, phage/polymer complexes carrying a therapeutic gene achieve greater cancer cell killing than phage alone. This new class of hybrid nanomaterial platform can advance targeted gene delivery applications by bacteriophage.

No MeSH data available.


Related in: MedlinePlus

Evaluation of the specificity of cell transduction by the targeted RGD4C-phage/polymer complexes. (a) Targeted gene transfer to 9L tumor cells by the RGD4C-phage/polymer was compared with that of negative phage/polymer controls (non-targeted or mutated). (b) Inhibition of transduction efficiency of the hybrid RGD4C-PDL and RGD4C-DEAE.DEX complexes by an RGD4C-expressing phage. (c) Targeted gene transfer by the RGD4C-phage/polymer complexes was assessed in the normal C2C12 myoblast cell line, using increasing concentrations of the PDL or DEAE.DEX cationic polymers. (d) Expression of the αv integrin receptor was investigated in 9L tumor cells and C2C12 myoblast cell line with anti-integrin primary and AlexasFluor-488 secondary antibodies.
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fig6: Evaluation of the specificity of cell transduction by the targeted RGD4C-phage/polymer complexes. (a) Targeted gene transfer to 9L tumor cells by the RGD4C-phage/polymer was compared with that of negative phage/polymer controls (non-targeted or mutated). (b) Inhibition of transduction efficiency of the hybrid RGD4C-PDL and RGD4C-DEAE.DEX complexes by an RGD4C-expressing phage. (c) Targeted gene transfer by the RGD4C-phage/polymer complexes was assessed in the normal C2C12 myoblast cell line, using increasing concentrations of the PDL or DEAE.DEX cationic polymers. (d) Expression of the αv integrin receptor was investigated in 9L tumor cells and C2C12 myoblast cell line with anti-integrin primary and AlexasFluor-488 secondary antibodies.

Mentions: We want to confirm that the targeting properties of the RGD4C-phage vector remain intact in the phage/polymer complex and that transduction of cells is specific and mediated by binding of the RGD4C ligand to the αv integrin receptors. Therefore, cell transduction efficiency of the RGD4C-phage/polymer complex was compared with that of complexes between polymer and either NT phage or phage displaying a mutant version of the RGD4C sequence (RGE4C, D–E), previously reported as NT controls for the RGD4C-phage.14,16 As shown in Figure 6a, Luc gene expression in cells treated with the RGE4C-phage/polymer was nonsignificant and comparable to that of NT phage/polymer complex.


Hybrid Nanomaterial Complexes for Advanced Phage-guided Gene Delivery.

Yata T, Lee KY, Dharakul T, Songsivilai S, Bismarck A, Mintz PJ, Hajitou A - Mol Ther Nucleic Acids (2014)

Evaluation of the specificity of cell transduction by the targeted RGD4C-phage/polymer complexes. (a) Targeted gene transfer to 9L tumor cells by the RGD4C-phage/polymer was compared with that of negative phage/polymer controls (non-targeted or mutated). (b) Inhibition of transduction efficiency of the hybrid RGD4C-PDL and RGD4C-DEAE.DEX complexes by an RGD4C-expressing phage. (c) Targeted gene transfer by the RGD4C-phage/polymer complexes was assessed in the normal C2C12 myoblast cell line, using increasing concentrations of the PDL or DEAE.DEX cationic polymers. (d) Expression of the αv integrin receptor was investigated in 9L tumor cells and C2C12 myoblast cell line with anti-integrin primary and AlexasFluor-488 secondary antibodies.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4221597&req=5

fig6: Evaluation of the specificity of cell transduction by the targeted RGD4C-phage/polymer complexes. (a) Targeted gene transfer to 9L tumor cells by the RGD4C-phage/polymer was compared with that of negative phage/polymer controls (non-targeted or mutated). (b) Inhibition of transduction efficiency of the hybrid RGD4C-PDL and RGD4C-DEAE.DEX complexes by an RGD4C-expressing phage. (c) Targeted gene transfer by the RGD4C-phage/polymer complexes was assessed in the normal C2C12 myoblast cell line, using increasing concentrations of the PDL or DEAE.DEX cationic polymers. (d) Expression of the αv integrin receptor was investigated in 9L tumor cells and C2C12 myoblast cell line with anti-integrin primary and AlexasFluor-488 secondary antibodies.
Mentions: We want to confirm that the targeting properties of the RGD4C-phage vector remain intact in the phage/polymer complex and that transduction of cells is specific and mediated by binding of the RGD4C ligand to the αv integrin receptors. Therefore, cell transduction efficiency of the RGD4C-phage/polymer complex was compared with that of complexes between polymer and either NT phage or phage displaying a mutant version of the RGD4C sequence (RGE4C, D–E), previously reported as NT controls for the RGD4C-phage.14,16 As shown in Figure 6a, Luc gene expression in cells treated with the RGE4C-phage/polymer was nonsignificant and comparable to that of NT phage/polymer complex.

Bottom Line: We demonstrate that the phage complex with cationic polymers generates positively charged phage and large aggregates that show enhanced cell surface attachment, buffering capacity, and improved transgene expression while retaining cell type specificity.Moreover, phage/polymer complexes carrying a therapeutic gene achieve greater cancer cell killing than phage alone.This new class of hybrid nanomaterial platform can advance targeted gene delivery applications by bacteriophage.

View Article: PubMed Central - PubMed

Affiliation: Phage Therapy Group, Department of Medicine, Imperial College London, London, UK.

ABSTRACT
Developing nanomaterials that are effective, safe, and selective for gene transfer applications is challenging. Bacteriophages (phage), viruses that infect bacteria only, have shown promise for targeted gene transfer applications. Unfortunately, limited progress has been achieved in improving their potential to overcome mammalian cellular barriers. We hypothesized that chemical modification of the bacteriophage capsid could be applied to improve targeted gene delivery by phage vectors into mammalian cells. Here, we introduce a novel hybrid system consisting of two classes of nanomaterial systems, cationic polymers and M13 bacteriophage virus particles genetically engineered to display a tumor-targeting ligand and carry a transgene cassette. We demonstrate that the phage complex with cationic polymers generates positively charged phage and large aggregates that show enhanced cell surface attachment, buffering capacity, and improved transgene expression while retaining cell type specificity. Moreover, phage/polymer complexes carrying a therapeutic gene achieve greater cancer cell killing than phage alone. This new class of hybrid nanomaterial platform can advance targeted gene delivery applications by bacteriophage.

No MeSH data available.


Related in: MedlinePlus