Peptide-Functionalized Dendrimer Nanocarriers for Targeted Microdystrophin Gene Delivery
Jessica Hersh,
José Manuel Condor Capcha,
Camila Iansen Irion,
Guerline Lambert,
Mauricio Noguera,
Mohit Singh,
Avinash Kaur,
Emre Dikici,
Joaquín J. Jiménez,
Lina A. Shehadeh,
Sylvia Daunert,
Sapna K. Deo
Affiliations
Jessica Hersh
Department of Biochemistry and Molecular Biology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
José Manuel Condor Capcha
Interdisciplinary Stem Cell Institute and Division of Cardiology, Department of Medicine, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Camila Iansen Irion
Interdisciplinary Stem Cell Institute and Division of Cardiology, Department of Medicine, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Guerline Lambert
Interdisciplinary Stem Cell Institute and Division of Cardiology, Department of Medicine, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Mauricio Noguera
Department of Biochemistry and Molecular Biology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Mohit Singh
Department of Biochemistry and Molecular Biology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Avinash Kaur
Department of Biochemistry and Molecular Biology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Emre Dikici
Department of Biochemistry and Molecular Biology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Joaquín J. Jiménez
Department of Biochemistry and Molecular Biology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Lina A. Shehadeh
Interdisciplinary Stem Cell Institute and Division of Cardiology, Department of Medicine, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Sylvia Daunert
Department of Biochemistry and Molecular Biology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
Sapna K. Deo
Department of Biochemistry and Molecular Biology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL 33136, USA
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, mdx4Cv. 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.