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A Multiple siRNA-Based Anti-HIV/SHIV Microbicide Shows Protection in Both In Vitro and In Vivo Models.

Boyapalle S, Xu W, Raulji P, Mohapatra S, Mohapatra SS - PLoS ONE (2015)

Bottom Line: Herein, we report the development of multifunctional chitosan-lipid nanocomplexes that can effectively deliver plasmids encoding siRNA(s) as microbicides without adverse effects and provide significant protection against HIV in both in vitro and in vivo models.Chitosan or chitosan-lipid (chlipid) was complexed with a cocktail of plasmids encoding HIV-1-specific siRNAs (psiRNAs) and evaluated for their efficacy in HEK-293 cells, PBMCs derived from nonhuman primates, 3-dimensional human vaginal ectocervical tissue (3D-VEC) model and also in non-human primate model.Moreover, prophylactic administration of the chlipid to deliver a psiRNA cocktail intravaginally with a cream formulation in a non-human primate model showed substantial reduction of SHIV (simian/human immunodeficiency virus SF162) viral titers.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine -Division of Translational Medicine and Nanomedicine Research Center, University of South Florida, Tampa, Florida, United States of America; Transgenex Nanobiotech Inc, Tampa, Florida, United States of America.

ABSTRACT
Human immunodeficiency virus (HIV) types 1 and 2 (HIV-1 and HIV-2) are the etiologic agents of AIDS. Most HIV-1 infected individuals worldwide are women, who acquire HIV infections during sexual contact. Blocking HIV mucosal transmission and local spread in the female lower genital tract is important in preventing infection and ultimately eliminating the pandemic. Microbicides work by destroying the microbes or preventing them from establishing an infection. Thus, a number of different types of microbicides are under investigation, however, the lack of their solubility and bioavailability, and toxicity has been major hurdles. Herein, we report the development of multifunctional chitosan-lipid nanocomplexes that can effectively deliver plasmids encoding siRNA(s) as microbicides without adverse effects and provide significant protection against HIV in both in vitro and in vivo models. Chitosan or chitosan-lipid (chlipid) was complexed with a cocktail of plasmids encoding HIV-1-specific siRNAs (psiRNAs) and evaluated for their efficacy in HEK-293 cells, PBMCs derived from nonhuman primates, 3-dimensional human vaginal ectocervical tissue (3D-VEC) model and also in non-human primate model. Moreover, prophylactic administration of the chlipid to deliver a psiRNA cocktail intravaginally with a cream formulation in a non-human primate model showed substantial reduction of SHIV (simian/human immunodeficiency virus SF162) viral titers. Taken together, these studies demonstrate the potential of chlipid-siRNA nanocomplexes as a potential genetic microbicide against HIV infections.

No MeSH data available.


Related in: MedlinePlus

Efficacy and safety of chlipids.(A) Transmission electron micrographs of chitosan, Lipofectamine and CNs complexed with pEGFP. (B) Distribution and quantification of in vivo transfection of pEGFP in BAL cells of C57/BL6 mice. pG: Naked EGFP plasmid; C+pG: chitosan-complexed pG; L+pG: Lipofectamine-complexed pG; CN+pG: chlipid-complexed pG. (C) Quantification of IL-6 in BAL following intranasal administration of nanoparticles. C: chitosan; L: Lipofectamine; CN: chlipid; C+pV: chitosan-complexed pVAX DNA; L+pV: Lipofectamine-complexed pVAX DNA; and CN+pV: chlipid-complexed pVAX DNA.
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pone.0135288.g001: Efficacy and safety of chlipids.(A) Transmission electron micrographs of chitosan, Lipofectamine and CNs complexed with pEGFP. (B) Distribution and quantification of in vivo transfection of pEGFP in BAL cells of C57/BL6 mice. pG: Naked EGFP plasmid; C+pG: chitosan-complexed pG; L+pG: Lipofectamine-complexed pG; CN+pG: chlipid-complexed pG. (C) Quantification of IL-6 in BAL following intranasal administration of nanoparticles. C: chitosan; L: Lipofectamine; CN: chlipid; C+pV: chitosan-complexed pVAX DNA; L+pV: Lipofectamine-complexed pVAX DNA; and CN+pV: chlipid-complexed pVAX DNA.

Mentions: Chitosan (1 mg/ml) and Lipofectamine 2000 (Invitrogen) were mixed at a ratio of 2:1 to form typical CNs and were viewed under an electron microscope. This suggested that CNs had a smaller size (441 nm) compared to the parent chitosan polymer (~1000 nm) (Fig 1A). Further, CNs were evaluated for their transfection efficiency in vivo using a mouse lung lavage assay. Plasmid encoding EGFP was complexed with chitosan, Lipofectamine or CNs and administered intranasally and 72 h later BAL cells were collected and GFP expression in the pooled lung lavage were examined by flow cytometry. CNs showed 30% transfection efficiency, as opposed to 20% with classical chitosan or Lipofectamine, and only 10% with naked DNA (Fig 1B). Moreover, very little IL-6 was found in the BAL of CN-complexed pEGFP group compared to chitosan-complexed group (Fig 1C).


A Multiple siRNA-Based Anti-HIV/SHIV Microbicide Shows Protection in Both In Vitro and In Vivo Models.

Boyapalle S, Xu W, Raulji P, Mohapatra S, Mohapatra SS - PLoS ONE (2015)

Efficacy and safety of chlipids.(A) Transmission electron micrographs of chitosan, Lipofectamine and CNs complexed with pEGFP. (B) Distribution and quantification of in vivo transfection of pEGFP in BAL cells of C57/BL6 mice. pG: Naked EGFP plasmid; C+pG: chitosan-complexed pG; L+pG: Lipofectamine-complexed pG; CN+pG: chlipid-complexed pG. (C) Quantification of IL-6 in BAL following intranasal administration of nanoparticles. C: chitosan; L: Lipofectamine; CN: chlipid; C+pV: chitosan-complexed pVAX DNA; L+pV: Lipofectamine-complexed pVAX DNA; and CN+pV: chlipid-complexed pVAX DNA.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4583459&req=5

pone.0135288.g001: Efficacy and safety of chlipids.(A) Transmission electron micrographs of chitosan, Lipofectamine and CNs complexed with pEGFP. (B) Distribution and quantification of in vivo transfection of pEGFP in BAL cells of C57/BL6 mice. pG: Naked EGFP plasmid; C+pG: chitosan-complexed pG; L+pG: Lipofectamine-complexed pG; CN+pG: chlipid-complexed pG. (C) Quantification of IL-6 in BAL following intranasal administration of nanoparticles. C: chitosan; L: Lipofectamine; CN: chlipid; C+pV: chitosan-complexed pVAX DNA; L+pV: Lipofectamine-complexed pVAX DNA; and CN+pV: chlipid-complexed pVAX DNA.
Mentions: Chitosan (1 mg/ml) and Lipofectamine 2000 (Invitrogen) were mixed at a ratio of 2:1 to form typical CNs and were viewed under an electron microscope. This suggested that CNs had a smaller size (441 nm) compared to the parent chitosan polymer (~1000 nm) (Fig 1A). Further, CNs were evaluated for their transfection efficiency in vivo using a mouse lung lavage assay. Plasmid encoding EGFP was complexed with chitosan, Lipofectamine or CNs and administered intranasally and 72 h later BAL cells were collected and GFP expression in the pooled lung lavage were examined by flow cytometry. CNs showed 30% transfection efficiency, as opposed to 20% with classical chitosan or Lipofectamine, and only 10% with naked DNA (Fig 1B). Moreover, very little IL-6 was found in the BAL of CN-complexed pEGFP group compared to chitosan-complexed group (Fig 1C).

Bottom Line: Herein, we report the development of multifunctional chitosan-lipid nanocomplexes that can effectively deliver plasmids encoding siRNA(s) as microbicides without adverse effects and provide significant protection against HIV in both in vitro and in vivo models.Chitosan or chitosan-lipid (chlipid) was complexed with a cocktail of plasmids encoding HIV-1-specific siRNAs (psiRNAs) and evaluated for their efficacy in HEK-293 cells, PBMCs derived from nonhuman primates, 3-dimensional human vaginal ectocervical tissue (3D-VEC) model and also in non-human primate model.Moreover, prophylactic administration of the chlipid to deliver a psiRNA cocktail intravaginally with a cream formulation in a non-human primate model showed substantial reduction of SHIV (simian/human immunodeficiency virus SF162) viral titers.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine -Division of Translational Medicine and Nanomedicine Research Center, University of South Florida, Tampa, Florida, United States of America; Transgenex Nanobiotech Inc, Tampa, Florida, United States of America.

ABSTRACT
Human immunodeficiency virus (HIV) types 1 and 2 (HIV-1 and HIV-2) are the etiologic agents of AIDS. Most HIV-1 infected individuals worldwide are women, who acquire HIV infections during sexual contact. Blocking HIV mucosal transmission and local spread in the female lower genital tract is important in preventing infection and ultimately eliminating the pandemic. Microbicides work by destroying the microbes or preventing them from establishing an infection. Thus, a number of different types of microbicides are under investigation, however, the lack of their solubility and bioavailability, and toxicity has been major hurdles. Herein, we report the development of multifunctional chitosan-lipid nanocomplexes that can effectively deliver plasmids encoding siRNA(s) as microbicides without adverse effects and provide significant protection against HIV in both in vitro and in vivo models. Chitosan or chitosan-lipid (chlipid) was complexed with a cocktail of plasmids encoding HIV-1-specific siRNAs (psiRNAs) and evaluated for their efficacy in HEK-293 cells, PBMCs derived from nonhuman primates, 3-dimensional human vaginal ectocervical tissue (3D-VEC) model and also in non-human primate model. Moreover, prophylactic administration of the chlipid to deliver a psiRNA cocktail intravaginally with a cream formulation in a non-human primate model showed substantial reduction of SHIV (simian/human immunodeficiency virus SF162) viral titers. Taken together, these studies demonstrate the potential of chlipid-siRNA nanocomplexes as a potential genetic microbicide against HIV infections.

No MeSH data available.


Related in: MedlinePlus