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A Proteomics Approach to Investigate miR-153-3p and miR-205-5p Targets in Neuroblastoma Cells
Ketan S Patil1, Indranil Basak1, Ramavati Pal1
1Department of Biological Sciences, St. John's University, New York, NY, 11439, United States of America.
Abstract:
MicroRNAs are key regulators associated with numerous diseases. In HEK293 cells, miR-153-3p and miR-205-5p down-regulate alpha-synuclein (SNCA) and Leucine-rich repeat kinase 2 (LRRK2), two key proteins involved in Parkinson's disease (PD). We have used two-dimensional gel electrophoresis (2D-PAGE) coupled to mass spectrometry (MS) 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 cells and 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. Modulation of miR-153-3p resulted in seven up-regulated proteins and eight down-regulated proteins. miR-205 modulation resulted in twelve up-regulated proteins and six down-regulated proteins. Several of the proteins are associated with neuronal processes, including peroxiredoxin-2 and -4, cofilin-1, prefoldin 2, alpha-enolase, human nucleoside diphosphate kinase B (Nm23) and 14-3-3 protein epsilon. Many of the differentially expressed proteins are involved in diverse pathways including metabolism, neurotrophin signaling, actin cytoskeletal regulation, HIF-1 signaling and the proteasome indicating that miR-153-3p and miR-205-5p are involved in the regulation of a wide variety of biological processes in neuroblastoma cells.
Insights
MicroRNAs miR-153-3p and miR-205-5p regulate Parkinson's disease proteins SNCA and LRRK2. Proteomics identified novel protein targets involved in neuronal processes and diverse cellular pathways.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) are crucial regulators in cellular processes and disease pathogenesis.
- Specific miRNAs, miR-153-3p and miR-205-5p, are implicated in down-regulating alpha-synuclein (SNCA) and Leucine-rich repeat kinase 2 (LRRK2), key proteins in Parkinson's disease (PD).
Purpose of the Study:
- To identify novel protein targets regulated by miR-153-3p and miR-205-5p in neuronal cells.
- To investigate the broader biological pathways influenced by these microRNAs in the context of neuroblastoma cells.
Main Methods:
- Utilized two-dimensional gel electrophoresis (2D-PAGE) combined with mass spectrometry (MS) for comparative proteomics.
- Overexpressed and inhibited miR-153-3p and miR-205-5p in SH-SY5Y neuroblastoma cells.
- Quantified and analyzed protein abundance changes using statistical significance (p-value < 0.05).
Main Results:
- Identified 33 protein spots with significant abundance changes upon miRNA modulation.
- miR-153-3p modulation affected 7 up-regulated and 8 down-regulated proteins.
- miR-205 modulation affected 12 up-regulated and 6 down-regulated proteins.
- Several identified proteins, including peroxiredoxins, cofilin-1, and alpha-enolase, are involved in neuronal functions.
Conclusions:
- miR-153-3p and miR-205-5p regulate a diverse set of proteins beyond SNCA and LRRK2.
- These microRNAs influence critical cellular pathways such as metabolism, neurotrophin signaling, and cytoskeletal regulation.
- The findings suggest a broad role for miR-153-3p and miR-205-5p in neuronal biology and disease, potentially impacting Parkinson's disease pathogenesis.
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