Enhancement of chitosan-mediated gene delivery through combination with phiC31 integrase

Oliveira, Ana V.; Silva, Gabriela A.; Chung, Daniel C.

doi:10.1016/j.actbio.2015.01.013

Cited by 13 publications

(8 citation statements)

References 31 publications

(48 reference statements)

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“…Many researchers, including the authors of this study, have tried to improve CS transfection efficiency through chemical structure modifications. 6,7,9,11-13 In our previous study, we confirmed that pNNS conjugated to CS (pNNS-CS) improved the transfection efficiency of plasmid DNA (pDNA) in C2C12 myoblast cells. 13 Therefore, we proposed the use of pNNS-CS as a gene carrier in gene therapy for the treatment of cartilage defects.…”

Section: Introductionsupporting

confidence: 70%

“…The effect of CS nanoparticles carrying small interfering RNA (siRNA), microRNA (miRNA), or therapeutic genes on gene delivery has been studied both in vitro and in vivo . 6-11 However, the transfection efficiency of CS is low under physiological conditions. Many researchers, including the authors of this study, have tried to improve CS transfection efficiency through chemical structure modifications.…”

Section: Introductionmentioning

confidence: 99%

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Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model

Zhao

Wang

Jia

et al. 2019

Bone & Joint Research

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Objectives Previously, we reported the improved transfection efficiency of a plasmid DNA-chitosan (pDNA-CS) complex using a phosphorylatable nuclear localization signal-linked nucleic kinase substrate short peptide (pNNS) conjugated to chitosan (pNNS-CS). This study investigated the effects of pNNS-CS-mediated miR-140 and interleukin-1 receptor antagonist protein (IL-1Ra) gene transfection both in rabbit chondrocytes and a cartilage defect model. Methods The pBudCE4.1-miR-140, pBudCE4.1-IL-1Ra, and negative control pBudCE4.1 plasmids were constructed and combined with pNNS-CS to form pDNA/pNNS-CS complexes. These complexes were transfected into chondrocytes or injected into the knee joint cavity. Results High IL-1Ra and miR-140 expression levels were detected both in vitro and in vivo. In vitro , compared with the pBudCE4.1 group, the transgenic group presented with significantly increased chondrocyte proliferation and glycosaminoglycan (GAG) synthesis, as well as increased collagen type II alpha 1 chain (COL2A1), aggrecan (ACAN), and TIMP metallopeptidase inhibitor 1 (TIMP-1) levels. Nitric oxide (NO) synthesis was reduced, as were a disintegrin and metalloproteinase with thrombospondin type 1 motif 5 (ADAMTS-5) and matrix metalloproteinase (MMP)-13 levels. In vivo , the exogenous genes reduced the synovial fluid GAG and NO concentrations and the ADAMTS-5 and MMP-13 levels in cartilage. In contrast, COL2A1, ACAN, and TIMP-1 levels were increased, and the cartilage Mankin score was decreased in the transgenic group compared with the pBudCE4.1 group. Double gene combination produced greater efficacies than each single gene, both in vitro and in vivo . Conclusion This study suggests that pNNS-CS is a good candidate for treating cartilage defects via gene therapy, and that IL-1Ra in combination with miR-140 produces promising biological effects on cartilage defects. Cite this article : R. Zhao, S. Wang, L. Jia, Q. Li, J. Qiao, X. Peng. Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model. Bone Joint Res 2019;8:165–178. DOI: 10.1302/2046-3758.83.BJR-2018-0222.R1.

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Section: Introductionsupporting

confidence: 70%

Section: Introductionmentioning

confidence: 99%

Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model

Zhao

Wang

Jia

et al. 2019

Bone & Joint Research

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“…Parameters as N:P ratio, molecular weight, mixing technique, among others, have been widely investigated on CS-DNA complexes [30,[32][33][34], and described as factors that influence the binding affinity between chitosan and DNA, size and charge of polyplexes, cellular uptake and dissociation of DNA, and thus, transfection efficiency [35][36][37]. Based on our previous work [10][11][12][13][14], an amine to phosphate (N:P) ratio of 15:1 was chosen to promote the formation of positively charged polyplexes and avoid repulsion by the negative cell surface.…”

Section: Polyplex Formulations Display Properties Adequate For Gene Tmentioning

confidence: 99%

Human-derived NLS enhance the gene transfer efficiency of chitosan

Bitoque

Morais

Oliveira

et al. 2020

Preprint

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Nuclear import is considered one of the major limitations for non-viral gene delivery systems and the incorporation of nuclear localization signals (NLS) that mediate nuclear intake can be used as a strategy to enhance internalization of exogenous DNA. In this work, human-derived endogenous NLS peptides based on Insulin Growth FactorBinding Proteins (IGFBP), namely IGFBP-3 and IGFBP-5, were tested for their ability to improve nuclear translocation of genetic material by non-viral vectors. Several strategies were tested to determine their effect on chitosan mediated transfection efficiency: co-administration with polyplexes, co-complexation at the time of polyplex formation, and covalent ligation to chitosan. Our results show that co-complexation and covalent ligation of human-derived NLS peptides to chitosan polyplexes yields a 2-fold increase in transfection efficiency.These results indicate that the integration of IGFBP-NLS peptides into polyplexes has potential as a strategy to enhance the efficiency of non-viral vectors.

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“…They have characterized the nanoparticles and significantly improved the transfection and integration efficacy in human embryonic kidney cells. They achieved higher expression levels of small and large integrated transgenes for over 10 weeks post transfection (Oliveira et al, 2015 ). Ease of transfection into mammalian cells using reliable synthetic formulations and variety of nanoparticles makes the DNA-based integrating systems superior compared to their viral counterparts, in particular in terms of immunocompatibility and safety with genotoxicity perspective.…”

Section: Piggybac Transposon Systemmentioning

confidence: 99%

Neuroprotective therapies in glaucoma: II. Genetic nanotechnology tools

Nafissi

Földvári

2015

Front. Neurosci.

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Neurotrophic factor genome engineering could have many potential applications not only in the deeper understanding of neurodegenerative disorders but also in improved therapeutics. The fields of nanomedicine, regenerative medicine, and gene/cell-based therapy have been revolutionized by the development of safer and efficient non-viral technologies for gene delivery and genome editing with modern techniques for insertion of the neurotrophic factors into clinically relevant cells for a more sustained pharmaceutical effect. It has been suggested that the long-term expression of neurotrophic factors is the ultimate approach to prevent and/or treat neurodegenerative disorders such as glaucoma in patients who do not respond to available treatments or are at the progressive stage of the disease. Recent preclinical research suggests that novel neuroprotective gene and cell therapeutics could be promising approaches for both non-invasive neuroprotection and regenerative functions in the eye. Several progenitor and retinal cell types have been investigated as potential candidates for glaucoma neurotrophin therapy either as targets for gene therapy, options for cell replacement therapy, or as vehicles for gene delivery. Therefore, in parallel with deeper understanding of the specific protective effects of different neurotrophic factors and the potential therapeutic cell candidates for glaucoma neuroprotection, the development of non-invasive and highly specific gene delivery methods with safe and effective technologies to modify cell candidates for life-long neuroprotection in the eye is essential before investing in this field.

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Enhancement of chitosan-mediated gene delivery through combination with phiC31 integrase

Cited by 13 publications

References 31 publications

Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model

Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model

Human-derived NLS enhance the gene transfer efficiency of chitosan

Neuroprotective therapies in glaucoma: II. Genetic nanotechnology tools

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