151 Hypertension Increases Fibrosis After Skeletal Muscle Injury in Hypertensive Spontaneously Rats Molecular Therapy Volume 22, Supplement 1, May 2014 Copyright © The American Society of Gene & Cell[.]
CELL AND GENE THERAPY FOR MYOGENIC DISORDERS conferred by F9 More importantly, F9 not only enhanced the delivery of PMOs but also applied to other nucleic acid cargoes e.g 2’-O-methylphosphorothioate RNA (2’OMe RNA) and siRNA Longer-term study further established the safety and efficacy of F9 in enhancing different cargoes to muscle Our findings demonstrate the potential of formulation F9 as an effective and safe formulation for cargoes in DMD and other muscle-related disorders 150 Ex Vivo Gene Therapy of Duchenne Muscular Dystrophy Using iPSCs from Duchenne Muscular Dystrophy Patients Chantale Maltais,1 William-Édouard Gravel,1 Carl Lebel,1 Jacques P Tremblay.1 Molecular Medecine, Laval University, Québec, Canada Flow cytometry demonstrated 89% and 54% transduction efficiency of the defined cardiomyocytes by AAV6 and AAV8, respectively, at MOI 10,000 We are currently repeating the same set of studies using AAV9 and 10 to confirm the titers for AAV6 and Once an optimal titer is determined, a blinded experiment will be conducted to compare transduction efficiency of all major serotypes at once in iPSC-CM from both normal and DMD patients: AAV1, 2, 5, 6, 8, and 10 The knowledge gained in this study will provide much needed information for clinical translation Further, this study will demonstrate proof –of-principle the feasibility that drug-discovery assays can be developed using iPSC-derived cardiomyocytes as a biological reagent 149 Novel Formulation Enhances Nucleic Acid Cargoes Delivery To Muscle Duchenne Muscular Dystrophy is a hereditary myopathy characterized by a progressive muscular degeneration, due to the absence of dystrophin Among the possible therapies there is the autologous transplantation of myoblasts derived from induced pluripotent stem cells (hiPSCs) derived from the dystrophic patient himself Indeed, hiPSCs represents a renewable source of myogenic autologous cells and would thus not induce an immune response of the patient against the transplanted cells However, there is currently no effective and reproducible protocol of differentiation of hiPSCs into myoblasts that results in considerable transplantation success We aim to induce myogenesis of hiPSCs with recombinant transcription factor proteins fused with a cell penetrating peptide, i.e., Tat These recombinant proteins were produced in bacteria and purified Our results showed that these transcription factors were able to enter into mesenchymal-like cells The capacity of these recombinant proteins to induce a myogenic differentiation is currently under study When these therapeutic approaches will be established, they could be clinically applied to treat dystrophic patients 151 Hypertension Increases Fibrosis After Skeletal Muscle Injury in Hypertensive Spontaneously Rats Gang Han,1 Xianjun Gao,1 Qingsong Wang,1 Limin Cao,1 Ben Gu,1 Haifang Yin.1 Research Center of Basic Medical Science, Tianjin Medical University, Tianjin, China Roberta Sessa Stilhano,1 Leonardo Martins,1 Priscila Keiko Matsumoto,1 Sang Won Han.1 Biophysics, Federal University of Sao Paulo, Sao Paulo, Brazil Duchenne muscular dystrophy (DMD) is one of the most common and severe forms of muscular dystrophy, arising from mutations in the dystrophin gene that preclude the synthesis of functional protein Currently there is no treatment available in clinic Although antisense oligonucleotides (AOs) – mediated exon skipping approach has shown great promise for DMD patients from encouraging phase IIb clinical trials, the lack of clear success of recent GSK phase III trials further underline the importance of developing ways to further enhance the systemic delivery efficiency of current AOs To address this issue, in our current study, we screened a number of formulations, which have been extensively applied in clinic with high safety profiles, in mdx mice to identify effective and safe delivery systemis for AOs currently in clinical trials From local intramuscular screening in mdx mice, we identified four top candidates which can increase the exon skipping efficiency of phosphorodiamidate morpholino oligomer (PMO), with up to fold increase in dystrophin expression in tibialis anterior (TA) treated with formulation F9 compared with that of saline Subsequent systemic administration studies revealed that F9 can significantly enhance the exon skipping efficacy of PMOs across different dosing regimens Notably, approximately 6.5 fold higher level of dystrophin was restored with PMOs in the presence of formulation F9 compared to saline at different tested dosing regimens Further investigation indicated that the increased exon skipping efficiency of PMOs is attributed to the enhanced delivery of PMOs to muscle Introduction: Fibrosis is a major acquired sequel after muscle injury, especially in more severe injuries in which healing occurs slowly, which prevents the proper muscle contraction and can lead to chronic pain and muscle contractures Fibrosis is a necessary step for recovery of damaged tissue, but when this process is exacerbated, as often occurs in cases of deeper lesions, causes imperfect tissue regeneration Hypertension may enhance fibrosis in cardiac muscle Aim: The main objective of this project is to prove that SHR (Spontaneously hypertensive rat) has more fibrosis than W (Wistar) after skeletal muscle injury Methods: The laceration model of injury was performed using a scissor to cut partly the tibial muscle Animals were divided in the following groups: SHR_L (SHR with laceration), SHR_LS (SHR with laceration followed by suture), W_L (W with laceration) and W_LS (W with laceration followed by suture) After 30 days, animals were euthanized and the tibial muscles were removed for histological analysis The following parameters were analyzed: Injury’s area (Macro and microscopic), amount of adipocytes, fibrosis’s area and muscle’s weight Results: L group present more injury’s area and adipocytes when compared with LS group There is no difference between L and LS in terms of fibrosis’s area The W_LS presented 1.3-fold more injury’s area than SHR_LS (p