Modelling Duchenne muscular dystrophy in MYOD1-converted urine-derived cells treated with 3-deazaneplanocin A hydrochloride

Hotake Takizawa,Ken Inoue,Yoshitsugu Aoki,Yuko Hara,Akihiko Ishiyama,Mikio Hoshino,Sadafumi Suzuki,Yuko Shimizu-Motohashi,Taisuke Ohno,Hirofumi Komaki,Yoshitaka Mizobe,Shin’Ichi Takeda,Eri Takeshita

doi:10.1038/s41598-019-40421-z

Abstract

Duchenne muscular dystrophy (DMD) is a severe muscle disorder characterised by mutations in the DMD gene. Recently, we have completed a phase I study in Japan based on systemic administration of the morpholino antisense that is amenable to exon-53 skipping, successfully. However, to achieve the effective treatment of DMD, in vitro assays on patient muscle cells to screen drugs and patient eligibility before clinical trials are indispensable. Here, we report a novel MYOD1-converted, urine-derived cells (UDCs) as a novel DMD muscle cell model. We discovered that 3-deazaneplanocin A hydrochloride, a histone methyltransferase inhibitor, could significantly promote MYOGENIN expression and myotube differentiation. We also demonstrated that our system, based on UDCs from DMD patients, could be used successfully to evaluate exon-skipping drugs targeting DMD exons including 44, 50, 51, and 55. This new autologous UDC-based disease modelling could lead to the application of precision medicine for various muscle diseases.

Highlights

Duchenne muscular dystrophy (DMD) is a severe muscle disorder characterised by mutations in the DMD gene
AQP1, specific for renal proximal tubular cells, was expressed only in partially needle-shaped urine-derived cells (UDCs). These mRNA expression patterns suggest that UDCs with high proliferative ability originated from the renal epithelial or uroepithelial cells including renal proximal tubular cells, which is consistent with earlier studies[6,7,14,15]
We discovered that an epigenetic modification using deazaneplanocin A hydrochloride (DZNep) could significantly promote the cell lineage conversion of UDCs into myotubes

Summary

Introduction

Duchenne muscular dystrophy (DMD) is a severe muscle disorder characterised by mutations in the DMD gene. Antoury et al reported that using mRNA isolated from DMD patient urine, they could successfully detect exon deletions in the DMD gene and confirmed exon-skipping activity after treatment with eteplirsen, an ASO that was granted accelerated approval by the U.S Food and Drug Administration in September of 20164. Their findings of ASO-dependent exon-skipping activity in urine provides the first non-invasive evaluation of ASO efficacy during a clinical trial. Kim et al demonstrated the direct-reprogramming of UDCs derived from limb-girdle muscular dystrophy and DMD patients to myogenic cells expressing MYOD1 using a lentiviral vector.

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