مطالعه بيوانفورماتيكي و آزمايشگاهي بيان RNAهاي غيركدكننده در سلول‌هاي عصبي حاصل از تمايز سلول‌هاي بنيادي

Expression variation of long noncoding RNAs in dopaminergic cells-derived from stem cells an‎d their MPP+ induced PD models

زيست شناسي گياهي و جانوري

Parkinsonʹs disease (PD) is one of the most common neurodegenerative disorders. Producing cells resembling dopaminergic neurons from stem cells is a promising approach for managing an‎d treating PD. Recently, it has been suggested that non-coding RNAs (ncRNAs), including lncRNAs an‎d microRNAs, play essential regulatory roles in various cellular processes such as differentiation an‎d brain development. Evidence indicates that lncRNAs often exhibit distinct expression patterns in the brain an‎d are involved in several neurological disorders an‎d biological functions. Weighted Gene Co-expression Network Analysis (WGCNA) can aid in identifying an‎d predicting key genes an‎d co-expressed genes involved in various biological processes. In this study, during the bioinformatics phase, a gene co-expression network was constructed for scRNA-seq gene expression data in dopaminergic neurons of the adult mouse substantia nigra. By examining the generated modules, key genes were identified. In the laboratory phase, the P19 embryonal carcinoma stem cell line was used as an in vitro model after differentiation induction into dopaminergic cells, assessing the expression of lncRNAs Malat1, Norad, Snhg1, an‎d Meg3. P19 cells were cultured under laboratory conditions an‎d then treated for 12 days with three groups: 1) total brain extract, 2) midbrain extract, an‎d 3) differentiation factors SHH, FGF, an‎d bFGF. Group 3 cells were treated with MPP+ for 24 hours to create a Parkinsonian-like group. The morphology of neurons in differentiated cells was confirmed through direct observations using cresyl violet staining an‎d light microscopy. The expression of specific neural proteins in differentiated P19 cells was examined using immunofluorescence an‎d flow cytometry. Results showed that after 12 days of differentiation protocol, neuronal phenotypes an‎d neuron-like cells were observable in all groups. Additionally, the eva‎luation of samples using immunofluorescence an‎d flow cytometry indicated that all these factors were significantly activated in all treated groups. Finally, comparative expression levels of lncRNAs Meg3, Norad, an‎d Snhg1 in the Parkinsonian-like group were significantly lower than those in the control group an‎d other differentiation groups. Conversely, the level of Malat1 was considerably higher compared to the control group. This research demonstrates that the expression levels of lncRNAs can reveal significant differences between differentiated dopaminergic cells an‎d their Parkinsonian models. The findings suggest that lncRNA biomarkers could serve as potential new therapeutic targets for PD. Furthermore, the reduction of Malat1 expression may have a protective effect on MPP+-induced PD models, thereby enhancing our understan‎ding of the molecular pathogenesis of PD.

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