Peptide Dendrimer/Lipid Hybrid Systems Are Efficient DNA Transfection Reagents: Structure–Activity Relationships Highlight the Role of Charge Distribution Across Dendrimer Generations

Kwok, Albert; Eggimann, Gabriela A.; Reymond, Jean‐Louis; Darbre, Tamis; Hollfelder, Florian

doi:10.1021/nn400343z

Cited by 85 publications

(77 citation statements)

References 99 publications

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“…For these experiments, the pH 4.7 and sMUA AuPAMAM vectors complexed with DNA were evaluated in comparison with a control and PEI/ DNA complexes. AlamarBlue assays showed some decrease in viability by PEI which is in agreement with other published results at the chosen N:P of 7.5 [39], and minimal cell death by the AuPAMAM/DNA complexes in SkBr3 cells (Fig. 8).…”

Section: Resultssupporting

confidence: 92%

Optimization of PAMAM-gold nanoparticle conjugation for gene therapy

et al. 2014

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The development of efficient and biocompatible non-viral vectors for gene therapy remains a great challenge, and exploiting the properties of both nanoparticle carriers and cationic polymers is an attractive approach. In this work, we have developed gold nanoparticle (AuNP) polyamidoamine (PAMAM) conjugates for use as non-viral transfection agents. AuPAMAM conjugates were prepared by crosslinking PAMAM dendrimers to carboxylic-terminated AuNPs via EDC and sulfo-NHS chemistry. EDC and sulfo-NHS have been utilized widely and in numerous applications such as amino acid coupling; however, their use in the coupling of PAMAM dendrimers to AuNPs presents new challenges to form effective and stable constructs for delivery that have not yet been examined. Enhanced colloidal stability and DNA condensation ability was established by probing two critical synthetic parameters: the reaction rate of the PAMAM crosslinking step, and the amine to carboxyl ratio. Based on this work, increasing the amine to carboxyl ratio during conjugation of PAMAM onto AuNPs yielded the optimal vector with respect to colloidal stability and transfection efficiency in vitro. AuPAMAM conjugates present attractive candidates for non-viral gene delivery due to their commercial availability, ease of fabrication and scale-up, high yield, high transfection efficiency and low cytotoxicity.

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Section: Resultssupporting

confidence: 92%

Optimization of PAMAM-gold nanoparticle conjugation for gene therapy

et al. 2014

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“…In addition, some dendrons and dendrimers, have shown intrinsic additional antitumoural properties, such as antiangiogenic actions [41], which makes them suitable molecules as scaffolds for antitumoural therapy. In addition, recent studies on peptide dendrimer/lipid mixtures used as effective transfection reagents also revealed the importance of appropriate charge distribution throughout the dendrimer tree, of the presence of active residues on the membrane/water interface, of the overall amphipathic character of the dendrimer and of the presence of co-lipids [42]. …”

Section: Discussionmentioning

confidence: 99%

Second Generation Amphiphilic Poly-Lysine Dendrons Inhibit Glioblastoma Cell Proliferation without Toxicity for Neurons or Astrocytes

et al. 2016

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Glioblastomas are the most common malignant primary brain tumours in adults and one of the most aggressive and difficult-to-treat cancers. No effective treatment exits actually for this tumour and new therapeutic approaches are needed for this disease. One possible innovative approach involves the nanoparticle-mediated specific delivery of drugs and/or genetic material to glioblastoma cells where they can provide therapeutic benefits. In the present work, we have synthesised and characterised several second generation amphiphilic polylysine dendrons to be used as siRNA carriers. We have found that, in addition to their siRNA binding properties, these new compounds inhibit the proliferation of two glioblastoma cell lines while being nontoxic for non-tumoural central nervous system cells like neurons and glia, cell types that share the anatomical space with glioblastoma cells during the course of the disease. The selective toxicity of these nanoparticles to glioblastoma cells, as compared to neurons and glial cells, involves mitochondrial depolarisation and reactive oxygen species production. This selective toxicity, together with the ability to complex and release siRNA, suggests that these new polylysine dendrons might offer a scaffold in the development of future nanoparticles designed to restrict the proliferation of glioblastoma cells.

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“…In vitro transfection studies have led to the discovery of potent gene transfer cationic lipids [3-5], polyethylenimine (PEI) and its analogues[6, 7], cyclodextrin derivatives[8, 9], chitosan and polysaccharide based polymers[10], dendrimers[11-13], polylysines[14, 15], and disulfide cross-linking polypeptides[16-18]. In vitro gene assays typically use plasmids expressing luciferase or green fluorescent protein (GFP) as reporter genes, which are most often performed in either 6-well or 96-well formats.…”

Section: Introductionmentioning

confidence: 99%

“…As chemical libraries grow, the miniaturization of gene transfer assays are necessary to better manage time, materials and cost. Gene transfer assays have been previously used in 384-well plate formats [6, 11, 19-27] but the full optimization of all parameters have not been reported and these have not been adapted to 1536 well plates [28-31]. In addition to transient transfection of cells, the use of cell lines stably transfected with luciferase or GFP have been used in a miniaturized assay format to facilitated screening for new siRNA delivery vectors [6, 11, 19-27].…”

Section: Introductionmentioning

confidence: 99%

Miniaturization of gene transfection assays in 384- and 1536-well microplates

Crowley

Duskey

et al. 2015

Analytical Biochemistry

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The miniaturization of gene transfer assays to either 384 or 1536-well plates greatly economizes the expense and allows much higher throughput when transfecting immortalized and primary cells compared to more conventional 96-well assays. To validate the approach, luciferase and GFP reporter gene transfer assays were developed to determine the influence of cell seeding number, transfection reagent to DNA ratios, transfection time, DNA dose, and luciferin dose on linearity and sensitivity. HepG2, CHO and 3T3 cells were transfected with polyethylenimine (PEI)-DNA in both 384 and 1536 well plates. The results established optimal transfection parameters in 384-well plates in an total assay volume of 35 μl and in 1536-well plates in a total volume of 8 μl. A luciferase assay performed in 384-well plates produced a Z' score of 0.53, making it acceptable for high throughput screening. Primary hepatocytes were harvested from mouse liver and transfected with PEI-DNA and calcium phosphate DNA nanoparticles in 384-well plates. Optimal transfection of primary hepatocytes was achieved on as few as 250 cells per well in 384-well plates, with CaPO4 proving to be 10-fold more potent than PEI.

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Peptide Dendrimer/Lipid Hybrid Systems Are Efficient DNA Transfection Reagents: Structure–Activity Relationships Highlight the Role of Charge Distribution Across Dendrimer Generations

Cited by 85 publications

References 99 publications

Optimization of PAMAM-gold nanoparticle conjugation for gene therapy

Optimization of PAMAM-gold nanoparticle conjugation for gene therapy

Second Generation Amphiphilic Poly-Lysine Dendrons Inhibit Glioblastoma Cell Proliferation without Toxicity for Neurons or Astrocytes

Miniaturization of gene transfection assays in 384- and 1536-well microplates

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