miR‐153‐3p And miR‐205‐5p Target Key Regulatory Pathways in Neuronal Cells

MicroRNAs are key regulators associated with a spectrum of diseases including neurodegenerative disorders. In non‐neuronal HEK293 cells and primary cultured cortical neurons, miR‐153‐3p and miR‐205‐5p down‐regulate a‐synuclein (SNCA) and Leucine‐rich repeat kinase 2 (LRRK2), two key proteins involved in Parkinson's disease (PD). In this study we have used two‐dimensional gel electrophoresis (2D‐PAGE) and mass spectrometry to identify a spectrum of miR‐153‐3p and miR‐205‐5p targets in neuronal SH‐SY5Y cells. We overexpressed and inhibited both microRNAs in SH‐SY5Y cellsand through comparative proteomics profiling we quantified ~240 protein spots from each analysis. Combined, thirty‐three protein spots were identified showing significant (p‐value < 0.05) changes in abundance. In response to altered levels of miR‐153‐3p seven proteins were up‐regulated and eight proteins were down‐regulated. In response to miR‐205‐5p modulation twelve proteins were up‐regulated whilst six proteins showed decreased abundance. Several of the proteins identified, including peroxiredoxin‐2, peroxiredoxin‐4, cofilin‐1, prefoldin 2, alpha‐enolase, human nucleoside diphosphate kinase B (Nm23) and 14‐3‐3 protein epsilon are associated with neuronal processes and are involved in diverse pathways including metabolism, neurotrophin signaling, actin cytoskeletal regulation, HIF‐1 signaling and the proteasome. Collectively, this demonstrates that miR‐153‐3p and miR‐205‐5p are associated with the regulation of a wide variety of biological processes in neurons.