Human sensory-like neuron surfaceome analysis

人类感觉样神经元表面组分析

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Abstract

Acral and triggerable pain is a hallmark of diseases involving small nerve fiber impairment, yet the underlying cellular mechanisms remain elusive. A key role is attributed to pain-related proteins located within the neuronal plasma membrane of nociceptive neurons. To explore this, we employed human induced pluripotent stem cell-derived sensory-like neurons and enriched their surface proteins by biotinylation. Samples from three independent cell differentiations were analyzed via liquid chromatography tandem mass spectrometry. Detected proteins were categorized by cellular location and function, followed by generating an interaction network for deregulated surface proteins. Gene expression of selected proteins was quantified using real-time PCR. A comparative analysis was performed between a patient with Fabry disease (FD) and a healthy control, which we used as model system. We successfully extracted surfaceome proteins from human sensory-like neurons, revealing deregulation of 48 surface proteins in FD-derived neurons. Among the candidates with potential involvement in pain pathophysiology were CACNA2D3, GPM6A, EGFR, and ABCA7. Despite the lack of gene expression differences in these candidates, the interaction network indicated compromised neuronal network integrity. Our approach successfully enabled the extraction and comprehensive analysis of the surfaceome from human sensory-like neurons, establishing a novel methodological framework for investigating human sensory-like neuron biology and cellular disease mechanisms.

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