Transcriptional comparison of human induced and primary midbrain dopaminergic neurons

Abstract

Generation of induced dopaminergic (iDA) neurons may provide a significant step forward towards cell replacement therapy for Parkinson’s disease (PD). To study and compare transcriptional programs of induced cells versus primary DA neurons is a preliminary step towards characterizing human iDA neurons. We have optimized a protocol to efficiently generate iDA neurons from human pluripotent stem cells (hPSCs). We then sequenced the transcriptomes of iDA neurons derived from 6 different hPSC lines and compared them to that of primary midbrain (mDA) neurons. We identified a small subset of genes with altered expression in derived iDA neurons from patients with Parkinson’s Disease (PD). We also observed that iDA neurons differ significantly from primary mDA neurons in global gene expression, especially in genes related to neuron maturation level. Results suggest iDA neurons from patient iPSCs could be useful for basic and translational studies, including in vitro modeling of PD. However, further refinement of methods of induction and maturation of neurons may better recapitulate full development of mDA neurons from hPSCs.

A novel floor plate boundary defined by adjacentEn1andDbx1microdomains distinguishes midbrain dopamine and hypothalamic neurons

“…However, immunolabeling studies on cultured and grafted DA neurons show that many neurons in the graft are FOXA2 + but not TH + , probably representing a non-DA fate. Gene expression analysis of these cultures showed robust upregulation of DBX1 and PITX2, in addition to DA markers (Kriks et al, 2011;Xia et al, 2016), indicative of imperfect programming. As these protocols were designed to maximize FOXA2 + /LMX1A + progenitor cells, our work would predict that both DBX1 and EN1 progenitors were generated, consequently resulting in the differentiation of DA and non-DA neurons.…”

Induction of Tyrosine Hydroxylase Gene Expression in Embryonal Carcinoma Stem Cells Using a Natural Tissue‐Specific Inducer

“…The results suggested some genes as new candidates in dopaminergic neuron differentiation for future studies. TH, Nr4a2, Slc6a3, Lmx1b, and Foxa2 genes are identified as well‐known DA markers (Poulin et al , ; Xia et al , ). The results of the present study indicated a significant co‐expression of Avp, Ache, Lhfpl5, and Dlk1 with both TH and Slc6a3 dopaminergic markers.…”

Cell therapy for central nervous system disorders: Current obstacles to progress

“…Inter‐clonal differences, nonhomogeneity of differentiation, and varied genetic backgrounds represent additional technical issues when priming iPS cell–derived differentiated neurons, oligodendrocytes, or astrocytes as cell products for CNS disorders. Gene expression of dopaminergic neurons derived from iPS cells of PD patients was reported to be significantly different from that of primary dopaminergic neurons . Beyond replacement of functional cells for patients with CNS disorders, iPS cells have broad potentials for establishing disease models that will be valuable for recapitulating pathological conditions and subsequent drug development.…”