Abstract
α-Synuclein (αSyn) is a small, disordered protein that becomes aggregated in Lewy body diseases, such as Parkinson’s disease (PD) and dementia with Lewy bodies (DLB). Human induced pluripotent stem cells (hiPSCs) potentially provide a tractable disease model to monitor early molecular changes associated with PD/DLB. We and others have previously derived hiPSC lines from patients with duplication and triplication of the SNCA gene, encoding for αSyn. Also, αSyn over-expression systems have been established in human embryonic stem cell (hESC) lines. A critical unresolved question is whether these pluripotent stem cell lines, with elevated levels of αSyn, can undergo efficient differentiation into dopaminergic and cortical neurons to model PD and DLB, respectively. It remains unclear if increased expression of αSyn affects hiPSC/hESC neural induction and neuronal differentiation. Therefore, we generated an allelic series of αSyn over-expressing hESC lines and characterised their potential for neurogenesis. Clonal hESC lines with significantly different levels of αSyn expression proliferated normally and maintained expression of pluripotent markers, such as OCT4. All cell lines efficiently produced PAX6+ neuroectoderm and there was no correlation between αSyn expression and neural induction efficiency. Finally, global transcriptomic analysis of cortical differentiation of hESC lines with low or high levels of αSyn expression demonstrated robust and similar induction of cortical neuronal expression profiles. Gene expression differences observed were unrelated to neural induction and neuronal differentiation. We conclude that elevated expression of αSyn in human pluripotent stem cells does not adversely affect their neuronal differentiation potential and this opens opportunities to model synucleinopathies including DLB.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
Conflicts of interest The authors have declared no competing interests.
Funding AN was funded by the Wellcome Trust Research Training Fellowship (203646/Z/16/Z). TK was funded by an MRC grant (MR/J012831/1).