Abstract
BackgroundDuchenne muscular dystrophy (DMD) is caused by mutations of the gene that encodes the protein dystrophin. A loss of dystrophin leads to severe and progressive muscle wasting in both skeletal and heart muscles. Human induced pluripotent stem cells (hiPSCs) and their derivatives offer important opportunities to treat a number of diseases. Here, we investigated whether givinostat (Givi), a histone deacetylase inhibitor, with muscle differentiation properties could reprogram hiPSCs into muscle progenitor cells (MPC) for DMD treatment.MethodsMPC were generated from hiPSCs by treatment with CHIR99021 and givinostat called Givi-MPC or with CHIR99021 and fibroblast growth factor as control-MPC. The proliferation and migration capacity were investigated by CCK-8, colony, and migration assays. Engraftment, pathological changes, and restoration of dystrophin were evaluated by in vivo transplantation of MPC. Conditioned medium from cultured MPC was collected and analyzed for extracellular vesicles (EVs).ResultsGivi-MPC exhibited superior proliferation and migration capacity compared to control-MPC. Givi-MPC produced less reactive oxygen species (ROS) after oxidative stress and insignificant expression of IL6 after TNF-α stimulation. Upon transplantation in cardiotoxin (CTX)-injured hind limb of Mdx/SCID mice, the Givi-MPC showed robust engraftment and restored dystrophin in the treated muscle than in those treated with control-MPC or human myoblasts. Givi-MPC significantly limited infiltration of inflammatory cells and reduced muscle necrosis and fibrosis. Additionally, Givi-MPC seeded the stem cell pool in the treated muscle. Moreover, EVs released from Givi-MPC were enriched in several miRNAs related to myoangiogenesis including miR-181a, miR-17, miR-210 and miR-107, and miR-19b compared with EVs from human myoblasts.ConclusionsIt is concluded that hiPSCs reprogrammed into MPC by givinostat possessing anti-oxidative, anti-inflammatory, and muscle gene-promoting properties effectively repaired injured muscle and restored dystrophin in the injured muscle.
Highlights
Duchenne muscular dystrophy (DMD) is caused by mutations of the gene that encodes the protein dystrophin
It is concluded that Human induced pluripotent stem cells (hiPSCs) reprogrammed into muscle progenitor cells (MPC) by givinostat possessing anti-oxidative, antiinflammatory, and muscle gene-promoting properties effectively repaired injured muscle and restored dystrophin in the injured muscle
Immunostaining revealed that the MPC derived from all iPSC lines were positive for the myogenic markers Pax7 and desmin (Fig. 1d) and the terminal differentiated cells expressed MF20 and displayed cell fusion (Fig. 1e), indicating their myogenic differentiation potential
Summary
Duchenne muscular dystrophy (DMD) is caused by mutations of the gene that encodes the protein dystrophin. Human induced pluripotent stem cells (hiPSCs) and their derivatives offer important opportunities to treat a number of diseases. We investigated whether givinostat (Givi), a histone deacetylase inhibitor, with muscle differentiation properties could reprogram hiPSCs into muscle progenitor cells (MPC) for DMD treatment. Human induced pluripotent stem cell (hiPSC)-derived derivatives offer important sources to treat a number of diseases. Efforts have been made in the past few years for the generation of muscle progenitor cells (MPC) from hiPSCs either by genetic modification or small molecules. It has been reported that MPC can be generated from teratoma with high engraftment efficiency in the muscle dystrophy model [10]. It seems more appropriate to look for alternate approaches for inducing MPC from hiPSCs with high engraftment and differentiation properties
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