Deriving Schwann Cells from hPSCs Enables Disease Modeling and Drug Discovery for Diabetic Peripheral Neuropathy

Abstract

Schwann cells (SCs) are the myelinating and non-myelinating glia of the peripheral nervous system (PNS) and are essential for its function. Defects in SCs are associated with many PNS disorders including diabetic peripheral neuropathy (DPN), a condition affecting millions of patients. Here we present a strategy for deriving and purifying SCs from human pluripotent stem cells (hPSCs). The scalable cultures of SCs allow basic and translational applications such as high-resolution molecular and functional characterization, developmental studies, modeling and mechanistic understanding of SC diseases and drug discovery. Our hPSC-derived SCs recapitulate the key molecular features of primary SCs and are capable of engrafting efficiently and producing myelin in injured sciatic nerves in rats. We further established an hPSC-based in vitro model of DPN that revealed the selective vulnerability of human SCs to hyperglycemia-induced cytotoxicity. We established a high-throughput screening system that identified a candidate drug that counteracts glucose-mediated cytotoxicity in SCs and normalizes glucose-induced transcriptional and metabolic abnormalities in SCs. Treatment of hyperglycemic mice with this drug candidate rescues sensory function, prevents SC death, and counteracts myelin damage in sciatic nerves suggesting considerable potential as a novel treatment for DPN.

Generation of Schwann cell derived melanocytes from hPSCs identifies pro-metastatic factors in melanoma

“…In agreement with our previous findings 16 , the Schwann cell master regulator EGR2 showed the highest level of expression and activity in S4 NCC2 compared to earlier S1-S3 populations and other S4 NCC subtypes (Figure 1F) . Interestingly, S4 NCC2 also expressed MSTN and SOX8 with high predicted SOX8 activity, both markers of NCCs transitioning to a SCP or “hub” transcriptional state recently described by Adameyko and colleagues 26,28 (Figure 1F) .…”

“…To systematically characterize how hPSC-NCC populations change over time, we performed single-cell RNA sequencing (scRNAseq) at five stages (S1-S5) of our previously established Schwann cell differentiation 16 . The timepoints sequenced were chosen to capture different stages of NCC specification and maturation: the earliest emergence of SOX10+ NCCs during the induction phase, an early, intermediate, and late stage of neural crest progression, and an early stage of glial induction (Figure 1A-B) .…”

“…The emerging S1 NCCs were homogenous, remaining as a single cluster, while S2, S3, and S4 NCCs contained two, three, and three subclusters, respectively (Figure 1D) . Stage 5 NCCs also contained three subclusters (Figure 1D) , which were annotated by comparison to our previously published 16 Schwann cell dataset from a similar differentiation stage. For this comparison we used similarity weighted non-negative embedding (SWNE) 25 .…”

“…Established strategies for differentiating NCCs from human pluripotent stem cell (hPSC) have provided an alternative model system with the ability to pattern into cranial, vagal, trunk and sacral regional identities [13][14][15] . Recently, Majd et al demonstrated temporal patterning of hPSC-NCCs with prolonged culture of 3D spheroids resulting in more efficient differentiation to glial lineages, mimicking the in vivo shift to gliogenesis in late-migrating NCCs 16 . This differentiation system enables the study of temporal fate restriction in NCCs, and promises access to lineages that emerge from late-migrating NCCs, such as parasympathetic neurons 17,18 .…”

CD146-assisted isolation and multi-omics characterisation of human pluripotent stem cell derived cranial Schwann cell precursor-like cells.

“…These data are consistent with previous observations made for NCC in the Ts65Dn mouse model of DS and reinforces the notion that defective Hedgehog signalling 160 is a likely contributor to the craniofacial defects associated with Down syndrome, as well as perhaps other features of DS such as Hirschsprung's disease, congenital heart defects, teeth and tongue abnormalities 20 , and altered skin pigmentation that likely involve SCPs. Collectively, the insights and datasets provided in this study should prove to be valuable resources for progressing investigations into the role of SCPs in normal and abnormal early human development, neurocristopathies, peripheral neuropathies 102,161 , and the pathogenesis of neuroblastoma 162 , and neurofibromatosis 23,26 .…”