Investigating SH-SY5Y Neuroblastoma Cell Surfaceome as a Model for Neuronal-Targeted Novel Therapeutic Modalities

Area of Science:

• Neuroscience
• Proteomics
• Drug Discovery

Background:

• SH-SY5Y cells are a key in vitro neuronal model for neurodegenerative diseases and pain.
• Their utility for receptor-mediated delivery of novel therapeutics, like antibody-drug conjugates, requires further study.
• Understanding the SH-SY5Y cell surfaceome enhances in vitro to in vivo correlation for accelerated drug discovery.

Purpose of the Study:

• To bioinformatically identify high-confidence neuronal surface proteins from brain and dorsal root ganglion (DRG) neurons.
• To delineate the SH-SY5Y cell surfaceome using surface proteomics.
• To compare the SH-SY5Y cell surfaceome with neuronal surface proteomes and investigate isoform-specific expression.

Main Methods:

• Developed a bioinformatic workflow using tissue proteomic and transcriptomic data to identify neuronal surface proteins.
• Performed surface proteomics on SH-SY5Y cells to define their cell surfaceome.
• Utilized comparative analysis and isoform-specific evaluation of proteomic data.

Main Results:

• The SH-SY5Y cell surfaceome significantly overlaps with the human brain and DRG neuronal surface proteome.
• Alternative splicing results in SH-SY5Y-specific major isoforms for 32% of common surface proteins, predicted to localize to the cell surface.
• Confirmed a SH-SY5Y-specific alternative NRCAM (neuron-glia related cell adhesion molecule) isoform, present in neuroblastomas but not typical brain neurons.

Conclusions:

• The SH-SY5Y cell surfaceome closely resembles that of brain and DRG neurons, validating its use as a neuronal model.
• Alternative splicing generates unique cell surface protein isoforms in SH-SY5Y cells, impacting drug targeting.
• The neuroblastoma-specific NRCAM isoform presents a potential target for neuroblastoma-specific therapeutics.