Mesenchymal stem cells derived from human induced pluripotent stem cells improve the engraftment of myogenic cells by secreting urokinase-type plasminogen activator receptor (uPAR)

Ahmed Elhussieny,Natsumi Takemura,Fusako Sakai-Takemura,Yusuke Maruyama,Wael Talaat Soliman,Ken’Ichiro Nogami,Yuko Miyagoe-Suzuki,Shin’Ichi Takeda

doi:10.1186/s13287-021-02594-1

Abstract

BackgroundDuchenne muscular dystrophy (DMD) is a severe X-linked recessive disease caused by mutations in the dystrophin gene. Transplantation of myogenic stem cells holds great promise for treating muscular dystrophies. However, poor engraftment of myogenic stem cells limits the therapeutic effects of cell therapy. Mesenchymal stem cells (MSCs) have been reported to secrete soluble factors necessary for skeletal muscle growth and regeneration.MethodsWe induced MSC-like cells (iMSCs) from induced pluripotent stem cells (iPSCs) and examined the effects of iMSCs on the proliferation and differentiation of human myogenic cells and on the engraftment of human myogenic cells in the tibialis anterior (TA) muscle of NSG-mdx4Cv mice, an immunodeficient dystrophin-deficient DMD model. We also examined the cytokines secreted by iMSCs and tested their effects on the engraftment of human myogenic cells.ResultsiMSCs promoted the proliferation and differentiation of human myogenic cells to the same extent as bone marrow-derived (BM)-MSCs in coculture experiments. In cell transplantation experiments, iMSCs significantly improved the engraftment of human myogenic cells injected into the TA muscle of NSG-mdx4Cv mice. Cytokine array analysis revealed that iMSCs produced insulin-like growth factor-binding protein 2 (IGFBP2), urokinase-type plasminogen activator receptor (uPAR), and brain-derived neurotrophic factor (BDNF) at higher levels than did BM-MSCs. We further found that uPAR stimulates the migration of human myogenic cells in vitro and promotes their engraftment into the TA muscles of immunodeficient NOD/Scid mice.ConclusionsOur results indicate that iMSCs are a new tool to improve the engraftment of myogenic progenitors in dystrophic muscle.

Highlights

Duchenne muscular dystrophy (DMD) is a devastating X-linked muscle disease that is caused by mutations in the DMD gene and affects 1 of 5000 male infants [1, 2].Elhussieny et al Stem Cell Res Ther (2021) 12:532Boys with DMD exhibit symptoms between 2 and 5 years of age, including delayed motor development, abnormal gait, and muscle weakness [2]
Positive staining with Alizarin red and Alcian blue in each differentiation condition further suggests the ability of Induced mesenchymal stem cells (iMSC) to differentiate into osteogenic and chondrogenic lineages, respectively (Fig. 1C)
IMSCs differentiated into oil red O-positive adipogenic cells with much less efficiency than bone marrow-derived (BM)-Mesenchymal stem cells (MSCs). iMSC cultures were negative for the pluripotency markers OCT3/4, SOX2, and NANOG by immunocytochemistry and RT-qPCR, indicating that there were no residual pluripotent stem cells in the culture (Additional file 1: Fig. 1C, D)

Summary

Introduction

Duchenne muscular dystrophy (DMD) is a devastating X-linked muscle disease that is caused by mutations in the DMD gene and affects 1 of 5000 male infants [1, 2].Elhussieny et al Stem Cell Res Ther (2021) 12:532Boys with DMD exhibit symptoms between 2 and 5 years of age, including delayed motor development, abnormal gait, and muscle weakness [2]. Duchenne muscular dystrophy (DMD) is a devastating X-linked muscle disease that is caused by mutations in the DMD gene and affects 1 of 5000 male infants [1, 2]. The DMD gene encodes the protein dystrophin. Transplantation of muscle stem/progenitor cells is a potential therapy for DMD, but the transplanted cells do not efficiently engraft and exert the therapeutic effects in the muscle of DMD patients [5]. Duchenne muscular dystrophy (DMD) is a severe X-linked recessive disease caused by mutations in the dystrophin gene. Transplantation of myogenic stem cells holds great promise for treating muscular dystrophies. Poor engraftment of myogenic stem cells limits the therapeutic effects of cell therapy. Mesenchymal stem cells (MSCs) have been reported to secrete soluble factors necessary for skeletal muscle growth and regeneration

Methods

Results

Discussion

Conclusion