Single-cell transcriptomics and In vitro lineage tracing reveals differential susceptibility of human iPSC-derived midbrain dopaminergic neurons in a cellular model of Parkinson's Disease

Fernandez Cardo, Lucia, Monzón Sandoval, Jimena, Li, Zongze ORCID: https://orcid.org/0000-0002-9923-7205, Webber, Caleb ORCID: https://orcid.org/0000-0001-8063-7674 and Li, Meng ORCID: https://orcid.org/0000-0002-4803-4643 2023. Single-cell transcriptomics and In vitro lineage tracing reveals differential susceptibility of human iPSC-derived midbrain dopaminergic neurons in a cellular model of Parkinson's Disease. Cells 12 (24) , 2860. 10.3390/cells12242860

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Abstract

Advances in stem cell technologies open up new avenues for modelling development and diseases. The success of these pursuits, however, relies on the use of cells most relevant to those targeted by the disease of interest, for example, midbrain dopaminergic neurons for Parkinson’s disease. In the present study, we report the generation of a human induced pluripotent stem cell (iPSC) line capable of purifying and tracing nascent midbrain dopaminergic progenitors and their differentiated progeny via the expression of a Blue Fluorescent Protein (BFP). This was achieved by CRISPR/Cas9-assisted knock-in of BFP and Cre into the safe harbour locus AAVS1 and an early midbrain dopaminergic lineage marker gene LMX1A, respectively. Immunocytochemical analysis and single-cell RNA sequencing of iPSC-derived neural cultures confirm developmental recapitulation of the human fetal midbrain and high-quality midbrain cells. By modelling Parkinson’s disease-related drug toxicity using 1-Methyl-4-phenylpyridinium (MPP+), we showed a preferential reduction of BFP+ cells, a finding demonstrated independently by cell death assays and single-cell transcriptomic analysis of MPP+ treated neural cultures. Together, these results highlight the importance of disease-relevant cell types in stem cell modelling.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	MRC Centre for Neuropsychiatric Genetics and Genomics (CNGG) Medicine Neuroscience and Mental Health Research Institute (NMHRI)
Publisher:	MDPI
ISSN:	2073-4409
Funders:	MRC
Date of First Compliant Deposit:	9 January 2024
Date of Acceptance:	15 December 2023
Last Modified:	28 Feb 2024 12:04
URI:	https://orca.cardiff.ac.uk/id/eprint/165374

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