Investigating SH-SY5Y Neuroblastoma Cell Surfaceome as a Model for Neuronal-Targeted Novel Therapeutic Modalities
Abstract
:1. Introduction
2. Results
2.1. Bioinformatically Inferring Surface Proteins Expressed in Human Brain Neurons and DRG Neurons Using Proteomics and Transcriptomics Data
2.2. Investigation of the SH-SY5Y Neuroblastoma Cell Surfaceome and Its Comparison to the Inferred Brain and DRG Neuronal Surfaceome
2.3. Validation of SH-SY5Y Surface Proteomics by RNA Sequencing and Identification of SH-SY5Y Specific Surface Protein Major Isoforms
2.4. SH-SY5Y Cells Specifically Express an Alternatively Spliced Isoform of NRCAM
3. Discussion
3.1. Surface Proteins Shared between Brain and DRG Neurons Show Tissue-Specific Differences in Expression Levels
3.2. Using the SH-SY5Y Neuroblastoma Cell Line as a Model for Receptor-Targeted Drug Delivery Needs to Be Evaluated on a Receptor-by-Receptor Basis
3.3. Investigation of Alternatively Spliced Surface Protein Isoforms in SH-SY5Y Cells Reveals NRCAM as a Putative Delivery Target for Neuroblastoma
4. Materials and Methods
4.1. Cell Culture
4.2. Cell Surface Protein Labeling and Immunoprecipitation
4.3. Sample Preparation for Mass Spectrometry Analysis
4.4. Mass Spectrometry Peptide Analysis
4.5. Mass Spectrometry Bioinformatic Analysis
4.6. Western Blot Analysis
4.6.1. Primary Antibodies Used
4.6.2. Secondary Antibodies Used
4.7. SH-SY5Y RNA Sequencing Sample Preparation and Data Analysis
4.8. SH-SY5Y RNA Sequencing Library Preparation, Sequencing, and Data Analysis
4.9. Major Isoform Calculation and Membrane Topology Benchmarking
4.10. Brain and DRG Transcriptomic and Proteomic Datasets
4.11. Brain and DRG (Dorsal Root Ganglion) Tissue Datasets Analysis
4.12. Surface Protein Selection
4.13. GO Term Enrichment Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gangras, P.; Gelfanova, V.; Williams, G.D.; Handelman, S.K.; Smith, R.M.; Debets, M.F. Investigating SH-SY5Y Neuroblastoma Cell Surfaceome as a Model for Neuronal-Targeted Novel Therapeutic Modalities. Int. J. Mol. Sci. 2022, 23, 15062. https://doi.org/10.3390/ijms232315062
Gangras P, Gelfanova V, Williams GD, Handelman SK, Smith RM, Debets MF. Investigating SH-SY5Y Neuroblastoma Cell Surfaceome as a Model for Neuronal-Targeted Novel Therapeutic Modalities. International Journal of Molecular Sciences. 2022; 23(23):15062. https://doi.org/10.3390/ijms232315062
Chicago/Turabian StyleGangras, Pooja, Valentina Gelfanova, Graham D. Williams, Samuel K. Handelman, Ryan M. Smith, and Marjoke F. Debets. 2022. "Investigating SH-SY5Y Neuroblastoma Cell Surfaceome as a Model for Neuronal-Targeted Novel Therapeutic Modalities" International Journal of Molecular Sciences 23, no. 23: 15062. https://doi.org/10.3390/ijms232315062