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Enhancing gene delivery of adeno-associated viruses by cell-permeable peptides.

Liu Y, Kim YJ, Ji M, Fang J, Siriwon N, Zhang LI, Wang P - Mol Ther Methods Clin Dev (2014)

Bottom Line: We show that CPPs increase internalization of viral particles into cells by facilitating both energy-independent and energy-dependent endocytosis.Moreover, CPPs can significantly enhance the endosomal escape process of viral particles, thus enhancing viral transduction to those cells that have exhibited very low permissiveness to AAV2 infection as a result of impaired intracellular viral processing.We also demonstrated that this approach could be applicable to other AAV serotypes.

View Article: PubMed Central - PubMed

Affiliation: Mork Family Department of Chemical Engineering and Materials Science, University of Southern California , Los Angeles, California, USA.

ABSTRACT
Adeno-associated virus type 2 (AAV2) is considered a promising gene delivery vector and has been extensively applied in several disease models; however, inefficient transduction in various cells and tissues has limited its widespread application in many areas of gene therapy. In this study, we have developed a general, but efficient, strategy to enhance viral transduction, both in vitro and in vivo, by incubating viral particles with cell-permeable peptides (CPPs). We show that CPPs increase internalization of viral particles into cells by facilitating both energy-independent and energy-dependent endocytosis. Moreover, CPPs can significantly enhance the endosomal escape process of viral particles, thus enhancing viral transduction to those cells that have exhibited very low permissiveness to AAV2 infection as a result of impaired intracellular viral processing. We also demonstrated that this approach could be applicable to other AAV serotypes. Thus, the membrane-penetrating ability of CPPs enables us to generate an efficient method for enhanced gene delivery of AAV vectors, potentially facilitating its applicability to human gene therapy.

No MeSH data available.


Related in: MedlinePlus

Cell-permeable peptides (CPPs) facilitate AAV8-mediated gene delivery in mouse muscles. (a,b) Histochemical and immunofluorescence analyses of muscle tissues from BALB/c mice injected with AAV8 or AAV8-CPP complexes. Muscle cross sections were taken at day 14 after intramuscular injection of AAV8 alone (108 particles/mice) or AAV8-CPP complexes. Muscle sections were stained with hematoxylin and eosin (a), or GFP antibody (red), followed by nuclear staining (blue, b). Scale bar represents 100 μm.
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fig10: Cell-permeable peptides (CPPs) facilitate AAV8-mediated gene delivery in mouse muscles. (a,b) Histochemical and immunofluorescence analyses of muscle tissues from BALB/c mice injected with AAV8 or AAV8-CPP complexes. Muscle cross sections were taken at day 14 after intramuscular injection of AAV8 alone (108 particles/mice) or AAV8-CPP complexes. Muscle sections were stained with hematoxylin and eosin (a), or GFP antibody (red), followed by nuclear staining (blue, b). Scale bar represents 100 μm.

Mentions: Moreover, we investigated the potential effects of the cell-permeable peptides on AAV8-mediated gene transfer in muscles. Complexes of AAV8 (108 particles) preincubated with or without CPPs (final concentration: 10 μmol/l) were injected intramuscularly into mice, and the GFP expression in muscles was analyzed 14 days postadministration. As shown in Figure 10a, no significant histopathologic change was observed for the muscle tissues from the mice treated with the AAV8-CPPs as compared to that treated with AAV8 alone, confirming that CPPs did not induce detectable cytotoxicity. More importantly, intramuscular injection of AAV8 in the presence of CPPs significantly increased the GFP expression in muscles as compared to that of AAV8 alone (Figure 10b). Thus, the fact that CPPs can enhance both AAV2 and AAV8 suggests that the approach to using CPP to improve gene delivery could be potentially applicable to various other AAV serotypes.


Enhancing gene delivery of adeno-associated viruses by cell-permeable peptides.

Liu Y, Kim YJ, Ji M, Fang J, Siriwon N, Zhang LI, Wang P - Mol Ther Methods Clin Dev (2014)

Cell-permeable peptides (CPPs) facilitate AAV8-mediated gene delivery in mouse muscles. (a,b) Histochemical and immunofluorescence analyses of muscle tissues from BALB/c mice injected with AAV8 or AAV8-CPP complexes. Muscle cross sections were taken at day 14 after intramuscular injection of AAV8 alone (108 particles/mice) or AAV8-CPP complexes. Muscle sections were stained with hematoxylin and eosin (a), or GFP antibody (red), followed by nuclear staining (blue, b). Scale bar represents 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4365833&req=5

fig10: Cell-permeable peptides (CPPs) facilitate AAV8-mediated gene delivery in mouse muscles. (a,b) Histochemical and immunofluorescence analyses of muscle tissues from BALB/c mice injected with AAV8 or AAV8-CPP complexes. Muscle cross sections were taken at day 14 after intramuscular injection of AAV8 alone (108 particles/mice) or AAV8-CPP complexes. Muscle sections were stained with hematoxylin and eosin (a), or GFP antibody (red), followed by nuclear staining (blue, b). Scale bar represents 100 μm.
Mentions: Moreover, we investigated the potential effects of the cell-permeable peptides on AAV8-mediated gene transfer in muscles. Complexes of AAV8 (108 particles) preincubated with or without CPPs (final concentration: 10 μmol/l) were injected intramuscularly into mice, and the GFP expression in muscles was analyzed 14 days postadministration. As shown in Figure 10a, no significant histopathologic change was observed for the muscle tissues from the mice treated with the AAV8-CPPs as compared to that treated with AAV8 alone, confirming that CPPs did not induce detectable cytotoxicity. More importantly, intramuscular injection of AAV8 in the presence of CPPs significantly increased the GFP expression in muscles as compared to that of AAV8 alone (Figure 10b). Thus, the fact that CPPs can enhance both AAV2 and AAV8 suggests that the approach to using CPP to improve gene delivery could be potentially applicable to various other AAV serotypes.

Bottom Line: We show that CPPs increase internalization of viral particles into cells by facilitating both energy-independent and energy-dependent endocytosis.Moreover, CPPs can significantly enhance the endosomal escape process of viral particles, thus enhancing viral transduction to those cells that have exhibited very low permissiveness to AAV2 infection as a result of impaired intracellular viral processing.We also demonstrated that this approach could be applicable to other AAV serotypes.

View Article: PubMed Central - PubMed

Affiliation: Mork Family Department of Chemical Engineering and Materials Science, University of Southern California , Los Angeles, California, USA.

ABSTRACT
Adeno-associated virus type 2 (AAV2) is considered a promising gene delivery vector and has been extensively applied in several disease models; however, inefficient transduction in various cells and tissues has limited its widespread application in many areas of gene therapy. In this study, we have developed a general, but efficient, strategy to enhance viral transduction, both in vitro and in vivo, by incubating viral particles with cell-permeable peptides (CPPs). We show that CPPs increase internalization of viral particles into cells by facilitating both energy-independent and energy-dependent endocytosis. Moreover, CPPs can significantly enhance the endosomal escape process of viral particles, thus enhancing viral transduction to those cells that have exhibited very low permissiveness to AAV2 infection as a result of impaired intracellular viral processing. We also demonstrated that this approach could be applicable to other AAV serotypes. Thus, the membrane-penetrating ability of CPPs enables us to generate an efficient method for enhanced gene delivery of AAV vectors, potentially facilitating its applicability to human gene therapy.

No MeSH data available.


Related in: MedlinePlus