Skeletal muscle cells derived from induced pluripotent stem cells: A platform for limb girdle muscular dystrophies

Abstract

Limb girdle muscular dystrophies (LGMD), caused by mutations in 29 different genes, are the fourth most prevalent group of genetic muscle diseases, leading to progressive weakness and atrophy of the skeletal muscles. Although the link between LGMD and their genetic origins has been determined, LGMD still represent an unmet medical need. In this paper, we describe a platform for modeling LGMD based on the use of human induced pluripotent stem cells (hiPSC). Thanks to the self-renewing and pluripotency properties of hiPSC, this platform provides an alternative and renewable source of skeletal muscle cells (skMC) to primary, immortalized or overexpressing cells. We report that skMC derived from hiPSC express the majority of the genes and proteins causing LGMD. As a proof of concept, we demonstrate the importance of this cellular model for studying LGMDR9 by evaluating disease-specific phenotypes in skMC derived from hiPSC obtained from four patients.

Pathological modeling of glycogen storage disease type III with CRISPR/Cas9 edited human pluripotent stem cells

“…In order to overcome these technical difficulties, hiPSCs have emerged as a relevant alternative. Indeed, the ease of reprogramming hiPSCs from blood samples or skin biopsies and recent advances in the development of skeletal myogenic differentiation protocols have allowed the generation of human skeletal muscle cell models suitable for the study of rare muscle diseases (Yoshida et al, 2017;Bruge et al, 2022;Guo et al, 2022;Laberthonnière et al, 2022;Ortuño-Costela et al, 2022;Shahriyari et al, 2022). Due to their unique self-renewal capacity, hiPSCs represent a standardized and unlimited source of cells, allowing the production of virtually infinite skeletal muscle cells and opening application for high-throughput drug screening.…”

“…GSDIII patient skMt were differentiated from hiPSCs, generated through the reprogramming of GSDIII patient fibroblasts (GM00576, Coriell Institute), which were characterized and differentiated by the same protocols as the GSDIII CRISPR and CTRL1 hiPSC lines (Supplementary Figures S2, S3). CTRL2 skMt were differentiated from a control hiPSCs characterized previously (Bruge et al, 2022) and differentiated by the same protocol (Supplementary Figure S3). Western blot analysis demonstrated the absence of GDE expression on both mutated skMt compared to controls (Figure 4A).…”

“…Since the discovery of human embryonic stem cells (hESCs) in 1998 (Thomson et al, 1998) and hiPSCs in 2006 (Takahashi and Yamanaka, 2006), effective differentiation protocols for an increasing number of cell types have been reported (Karagiannis et al, 2019). A large number of studies, including from our own group, have highlighted the use of hiPSCs for the pathological modeling of muscular diseases of genetic origin (Tedesco et al, 2012;Tanaka et al, 2013;Shoji et al, 2015;Wu et al, 2017;El-Battrawy et al, 2018;Mateos-Aierdi et al, 2021;Mérien et al, 2021;Bruge et al, 2022). In this study, we first established a stable AGL knockout hiPSC line using CRISPR/Cas9 gene editing technology (GSDIII CRISPR ).…”