Peptide-Functionalized Dendrimer Nanocarriers for Targeted Microdystrophin Gene Delivery.
Jessica HershJosé Manuel Condor CapchaCamila Iansen IrionGuerline LambertMauricio NogueraMohit SinghAvinash KaurEmre DikiciJoaquín J JiménezLina A ShehadehSylvia DaunertSapna K DeoPublished in: Pharmaceutics (2021)
Gene therapy is a good alternative for determined congenital disorders; however, there are numerous limitations for gene delivery in vivo including targeted cellular uptake, intracellular trafficking, and transport through the nuclear membrane. Here, a modified G5 polyamidoamine (G5 PAMAM) dendrimer-DNA complex was developed, which will allow cell-specific targeting to skeletal muscle cells and transport the DNA through the intracellular machinery and the nuclear membrane. The G5 PAMAM nanocarrier was modified with a skeletal muscle-targeting peptide (SMTP), a DLC8-binding peptide (DBP) for intracellular transport, and a nuclear localization signaling peptide (NLS) for nuclear uptake, and polyplexed with plasmid DNA containing the GFP-tagged microdystrophin ( µDys ) gene. The delivery of µDys has been considered as a therapeutic modality for patients suffering from a debilitating Duchenne muscular dystrophy (DMD) disorder. The nanocarrier-peptide-DNA polyplexes were prepared with different charge ratios and characterized for stability, size, surface charge, and cytotoxicity. Using the optimized nanocarrier polyplexes, the transfection efficiency in vitro was determined by demonstrating the expression of the GFP and the µDys protein using fluorescence and Western blotting studies, respectively. Protein expression in vivo was determined by injecting an optimal nanocarrier polyplex formulation to Duchenne model mice, mdx 4Cv . Ultimately, these nanocarrier polyplexes will allow targeted delivery of the microdystrophin gene to skeletal muscle cells and result in improved muscle function in Duchenne muscular dystrophy patients.
Keyphrases
- duchenne muscular dystrophy
- skeletal muscle
- drug delivery
- cancer therapy
- single molecule
- end stage renal disease
- circulating tumor
- induced apoptosis
- cell free
- newly diagnosed
- insulin resistance
- ejection fraction
- gene therapy
- chronic kidney disease
- peritoneal dialysis
- prognostic factors
- escherichia coli
- poor prognosis
- genome wide
- mesenchymal stem cells
- binding protein
- cell cycle arrest
- single cell
- gene expression
- signaling pathway
- type diabetes
- copy number
- quantum dots
- cell therapy
- cell death
- south africa
- metabolic syndrome
- mass spectrometry
- patient reported
- high fat diet induced
- solar cells