Comparison of three congruent patient-specific cell types for the modelling of a human genetic Schwann-cell disorder

Abstract

Patient-specific human induced pluripotent stem cells (hiPSCs) hold great promise for the modelling of genetic disorders. However, these cells display wide intra-individual and inter-individual variations in gene expression, making it challenging to distinguish true-positive and false-positive phenotypes. Also, data from hiPSC phenotypes and from human embryonic stem cells (hESCs) harbouring the same disease mutation are lacking. Here, we report a comparison of molecular, cellular and functional characteristics of three congruent patient-specific cell types ― hiPSCs, hESCs, and direct lineage-converted cells ― derived from currently available differentiation and direct-reprogramming technologies, for the modelling of Charcot Marie Tooth 1A, a human genetic Schwann-cell disorder featuring a 1.4 megabase chromosomal duplication. In particular, we find that the chemokines CXCL1 and MCP1 are commonly upregulated in all three congruent models and in clinical patient samples. The development of congruent models of a single genetic disease by using somatic cells from a common patient will facilitate the search for convergent phenotypes.

Inhibition of the Combinatorial Signaling of Transforming Growth Factor-Beta and NOTCH Promotes Myotube Formation of Human Pluripotent Stem Cell-Derived Skeletal Muscle Progenitor Cells

“…Furthermore, the inhibition of NOTCH signaling abolished the effect of TGFβ ( Figure 4 b,c,g,h), indicating that the myotube generating process is a target regulated by the combinatorial mechanism of TGFβ and NOTCH [ 22 ]. To corroborate the effect of the combinational inhibition of TGFβ and NOTCH on the promotion of myotube formation, we employed human primary myoblasts, another model system that could rule out biological or technical errors in studies with iPSCs [ 31 ]. The promotion of myotube formation by combinational inhibition of TGFβ and NOTCH was reproduced in the primary myoblasts-based experiment ( Figure 5 a–c), and furthermore, the effect on myotube formation was also demonstrated in iPSC-derived myoblasts from DMD patients ( Figure 5 f–j).…”

Neuromuscular Development and Disease: Learning From in vitro and in vivo Models

“…This method shortened the total differentiation time from approximately 41 days ( Huang et al, 2017 ) to approximately 32 days ( Kim et al, 2017 ). However, a more recent protocol has been developed for the derivation of direct Schwann-cell precursors (SCPs) from SOX10-reporting hiPSCs that only required a total differentiation time of 21 days and allowed for in vitro culture up to 80 days with maintained expression of the SC proteins S100b, glial fibrillary acidic protein (GFAP), and galactosylceramidase ( Mukherjee-Clavin et al, 2019 ). With all these methods, differentiated SCs show increased expression of SC-specific markers such as GFAP and S100β.…”

Schwann Cell Cultures: Biology, Technology and Therapeutics

“…Differentiation protocols and analysis of myelinating cells were described in [48]. The image of iSCs was kindly provided by Dr. Yong Jun Kim [49].…”

“…The concept of SC creation is rather new [56]. In this paradigm, cells with SC characteristics (iSCs) are derived by using directed in vitro differentiation (e.g., from multipotent neural or non-neural stem cells) [57], transdifferentiation [56] or even direct conversion from somatic cells such as fibroblasts [49,58]. iSCs can be generated from embryonic stem cells, induced pluripotent stem cells (iPSCs), [59], neural stem cells (e.g., neural crest-derived) and mesenchymal stem cells (MSCs) originating from the bone marrow [60][61][62] or adipose tissue [63].…”

“…This is due mainly to the prospect of obtaining human SCs without major ethical concerns. SC derivation from iPSCs offer an alternative, potentially unlimited source of patient-specific human SCs with broad applications in modeling genetic disorders that affect SC function such as Charcot-Marie-Tooth disease [49]. Yet, how much iSCs resemble the SCs from peripheral nerve or how stable they are as they are passaged or manipulated in culture remains to be further explored.…”