Generating graftable dopaminergic neurons by NR4A2 activation and exploring associated lncRNA signatures

Abstract

Non-coding RNAs (ncRNAs) are crucial in neural development and neurodegenerative diseases like Parkinson's disease (PD). Long non-coding RNAs (lncRNAs) such as Neat1, Hotair, and Uchl1os are pivotal for mitochondrial function and dopaminergic neuron viability. This study utilized weighted gene co-expression network analysis in the substantia nigra of Parkinson's patients to identify nine key genes related to dopaminergic neuron development, emphasizing the transcription factor NR4A2 for further analysis through overexpression in P19 cells. P19 stem cells were induced to differentiate into dopaminergic neurons prior to transplantation into a PD rat model. The expression levels of lncRNAs and dopaminergic markers were evaluated using real-time PCR in both in vitro and in vivo settings. Flow cytometry was employed to detect specific proteins in differentiated cells from midbrain extracts. Immunofluorescence, flow cytometry, and Western blot techniques confirmed the presence of dopaminergic cell-specific proteins. The results indicated an increase in the expression of lncRNAs Neat1, Hotair, and Uchl1os post-transplantation of differentiated cells into the PD rat model. Moreover, dopamine concentrations were elevated in the differentiated group relative to the control. The expression levels of dopaminergic markers and lncRNAs exhibited variability in both in vitro and in vivo conditions. In summary, the findings of this investigation reveal that stimulating NR4A2 in P19 cells promotes the production of dopaminergic neurons viable for grafting in experimental models of PD, coupled with unique lncRNA expression patterns featuring heightened Neat1, Hotair, and Uchl1os.