Donepezil promotes differentiation of neural stem cells into mature oligodendrocytes at the expense of astrogenesis

Abstract

Oligodendrocytes are the myelin-forming cells of the central nervous system. Oligodendrocyte loss and failure of myelin development result in serious human disorders, including multiple sclerosis. Previously, using oligodendrocyte progenitor cells, we have shown that donepezil, which is an acetylcholinesterase inhibitor developed for the treatment of Alzheimer's disease, stimulates myelin gene expression and oligodendrocyte differentiation. Here, we aimed to analyze the effects of donepezil on primary mouse embryonic neural stem cells (NSCs). Donepezil treatment led to impaired self-renewal ability and increased apoptosis. These effects appeared to be mediated through the Akt/Bad signaling pathway. Using neurosphere differentiation analysis, we observed that donepezil leads to reduced numbers of astrocytes and increased numbers of oligodendrocytes and neurons. Consistent with this finding, mRNA and protein levels for the oligodendrocyte markers myelin-associated glycoprotein, 2', 3'-cyclic-nucleotide 3'-phosphodiesterase (CNPase), and myelin basic protein, as well as the neuronal marker β-tubulin type III (Tuj1) were up-regulated. In contrast, the expression of the astrocyte marker glial fibrillary acidic protein (GFAP) was down-regulated by donepezil in a dose- and time-dependent manner. Moreover, donepezil increased oligodendrocyte differentiation, resulting in a reduction in the differentiation of NSCs into astrocytes, by suppressing the activation of signal transducer and activator of transcription 3 (STAT3), SMAD1/5/9, and the downstream target gene GFAP, even under astrocyte-inducing conditions. These results suggest that efficient differentiation of NSCs into oligodendrocytes by donepezil may indicate a novel therapeutic role for this drug in promoting repair in demyelinated lesions in addition to its role in preventing astrogenesis.

Donepezil‐induced oligodendrocyte differentiation is mediated through estrogen receptors

“…Western blotting revealed that the expression levels of myelin‐associated glycoprotein (MAG) and myelin basic protein (MBP) were enhanced by donepezil in a concentration‐dependent manner; considerable effect was observed at 10 μM, whereas the maximum effect was observed at 40 μM (Figure 1b). Consistent with previous reports, this result indicated that the relatively high doses of donepezil could successfully promote oligodendrocyte differentiation (Imamura et al, 2015, 2017). To determine the effect of donepezil on OPC specification and oligodendrocyte differentiation from miPSC‐NSCs, miPSC‐NSCs were differentiated with or without 20 μM donepezil in differentiation medium for 5 days.…”

“…Previously, we showed using primary mouse embryonic NSCs that donepezil also leads to increased numbers of neurons and reduced number of astrocytes (Imamura et al, 2017). Co‐immunostaining analysis revealed that ERs were co‐localized with both Tuj1, a neuronal marker, and GFAP, an astrocyte marker (Figure S2).…”

“…We discovered that donepezil, an approved drug for the treatment of Alzheimer's disease, significantly induces the expression of myelin-associated genes and stimulates the differentiation of OPCs into mature oligodendrocytes (Imamura et al, 2015). Additionally, donepezil was observed to significantly stimulate the differentiation of NSCs into mature oligodendrocytes and immature neurons under conditions of differentiation (Imamura, Arai, Dateki, & Takishima, 2017). In this study, we first investigated the effects of donepezil on the differentiation of NSCs derived from mouse iPSCs (miP-SC-NSCs).…”

Aberrant Oligodendrogenesis in Down Syndrome: Shift in Gliogenesis?

“…In this context, such a preferred shift toward oligodendroglia could be mediated via the acetylcholine esterase inhibitor donepezil, an FDA-approved drug for Alzheimer's disease and dementia. Donepezil was shown to promote the differentiation of primary NSCs into mature oligodendrocytes at the expense of astrocytes [150], and it was also found to enhance myelin sheath generation in neuron/glia co-cultures [151]. The FDA-approved anti-seizure drug ethosuximide was described to be capable of inducing trans-differentiation of muscle-derived stem cells into Olig2-positive oligodendroglial cells [152].…”

Insects in anthelminthics research: Lady beetle-derived harmonine affects survival, reproduction and stem cell proliferation of Schistosoma mansoni

“…For instance, embryonic stem cells of human and mouse express AChE and showed reduced proliferation upon ACh stimulation [67, 68], just like an ACh over-abundance by inhibition of AChE activity might do. Furthermore, inhibition of AChE by organophosphates in mesenchymal stem cells or by donezepil in neural stem cells led to suppression of proliferation, differentiation and self-renewal ability [41, 69]. Therefore, future studies should address the possible role of ACh signaling in schistosomal stem cells in detail, and we propose RT-qPCR analysis of sorted stem cells as a start.…”