Assessing and enhancing migration of human myogenic progenitors using directed iPS cell differentiation and advanced tissue modelling

Abstract

Muscle satellite stem cells (MuSCs) are responsible for skeletal muscle growth and regeneration. Despite their differentiation potential, human MuSCs have limited in vitro expansion and in vivo migration capacity, limiting their use in cell therapies for diseases affecting multiple skeletal muscles. Several protocols have been developed to derive MuSC-like progenitors from human induced pluripotent stem (iPS) cells (hiPSCs) to establish a source of myogenic cells with controllable proliferation and differentiation. However, current hiPSC myogenic derivatives also suffer from limitations of cell migration, ultimately delaying their clinical translation. Here we use a multidisciplinary approach including bioinformatics and tissue engineering to show that DLL4 and PDGF-BB improve migration of hiPSCderived myogenic progenitors. Transcriptomic analyses demonstrate that this property is conserved across species and multiple hiPSC lines, consistent with results from single cell motility profiling. Treated cells showed enhanced trans-endothelial migration in transwell assays. Finally, increased motility was detected in a novel humanised assay to study cell migration using 3D artificial muscles, harnessing advanced tissue modelling to move hiPSCs closer to future muscle gene and cell therapies.

CRISPR-Cas9 mediated endogenous utrophin upregulation improves Duchenne Muscular Dystrophy

“…We then assessed if the editing strategy would induce utrophin upregulation and correct sarcolemma localization in a model more closely resembling skeletal muscle tissue architecture than conventional monolayer cultures (65). To this aim, we generated 3D engineered skeletal muscles by culturing Cas9 only or Cas9/gRNA treated hDMD myoblasts in our established 3D platform capable to model muscular dystrophies and therapeutic interventions, including the possibility to study muscle function (65)(66)(67)(68)(69). 8 days after treatment, edited and control hDMD myoblasts were differentiated into a 3D hydrogel as previously described (69) (Fig 4A ; Fig S3A-B).…”

Effect of heme oxygenase-1 on the differentiation of human myoblasts and the regeneration of murine skeletal muscles after acute and chronic injury

“…This may indicate that hiPSC-SkM-HO-1 cells affected earlier stages of muscle regeneration. Problems with the survival and/or integration of myoblasts/in vivo engraftment have been shown to limit the application of this technology (for references, see: [ 28 ]). Therefore, modifications of established protocols to increase the number of myogenic progenitors and their regenerative potential in vivo seem to be crucial for a successful therapy.…”

“…It was shown that the overexpression of Pax7 transcription factor [ 29 ] or the inhibition of p38 signaling pathway in vitro during mSC expansion enhances the cell potential to engraft in vivo [ 30 ]. A recent study by Choi et al [ 28 ] suggested modulation of NOTCH and PDGF signalling as a way to increase the migration of hiPSC-derived muscle cells for more efficient muscle cell therapies. The absence of hiPSC-derived myoblasts in murine muscle tissue may be affected by the number of injected cells, their (immature) properties, or the timing of the experiment.…”

A self-renewing biomimetic skeletal muscle construct engineered using induced myogenic progenitor cells

“…It will be of interest to assess if a fraction of the Pax7 + cells in iSMCs exit the cell cycle and return to quiescence, as previously reported for SCs seeded on myofibers [56][57][58] . Additionally, it will be of interest to assess whether serum withdrawal or modulation of additional signaling pathways might confer quiescence rather than activation on the Pax7 + cell population in iSMCs, rendering them more similar to muscle tissue during homeostasis 57,59,60 .…”