Cheng Juan, Ji Dongrui, Ma Jing, Zhang Qinghua, Zhang Wanglin, Yang Lin
Department of Clinical Laboratory, Shanghai Children's Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Wayen Biotechnologies (Shanghai), Inc., Shanghai, China.
Front Oncol. 2024 Aug 20;14:1367159. doi: 10.3389/fonc.2024.1367159. eCollection 2024.
Neuroblastoma (NB) primarily arises in children who are <10 years of age, and originates from developing sympathetic nervous system, which results in tumors in adrenal glands and/or sympathetic ganglia. The diagnosis of NB involves a combination of laboratory and imaging tests, and biopsies. Small extracellular vesicles (sEVs) have gained attention as potential biomarkers for various types of tumors. Here, we performed proteomic analysis of serum sEVs and identified potential biomarkers for NB.
Label-free proteomics of serum sEVs were performed in the discovery phase. A bulk RNA-seq dataset of NB tissues was used to analyze the association between genes encoding sEVs proteins and prognosis. Potential biomarkers were validated via multiple reaction monitoring (MRM) or western blot analysis in the validation phase. A public single-cell RNA-seq (scRNA-seq) dataset was integrated to analyze the tissue origin of sEVs harboring biomarkers.
A total of 104 differentially expressed proteins were identified in NB patients with label-free proteomics, and 26 potential biomarkers were validated with MRM analysis. Seven proteins BSG, HSP90AB1, SLC44A1, CHGA, ATP6V0A1, ITGAL and SELL showed the strong ability to distinguish NB patients from healthy controls and non-NB patients as well. Integrated analysis of scRNA-seq and sEVs proteomics revealed that these sEVs-derived biomarkers originated from different cell populations in tumor tissues.
sEVs-based biomarkers may aid the molecular diagnosis of NB, representing an innovative strategy to improve NB detection and management.
神经母细胞瘤（NB）主要发生在10岁以下儿童，起源于发育中的交感神经系统，可导致肾上腺和/或交感神经节出现肿瘤。NB的诊断需要结合实验室检查、影像学检查和活检。小细胞外囊泡（sEVs）作为各种肿瘤的潜在生物标志物受到关注。在此，我们对血清sEVs进行了蛋白质组学分析，并鉴定了NB的潜在生物标志物。
在发现阶段对血清sEVs进行无标记蛋白质组学分析。利用NB组织的大量RNA测序数据集分析编码sEVs蛋白的基因与预后之间的关联。在验证阶段，通过多反应监测（MRM）或蛋白质印迹分析对潜在生物标志物进行验证。整合一个公共的单细胞RNA测序（scRNA-seq）数据集，以分析携带生物标志物的sEVs的组织来源。
通过无标记蛋白质组学在NB患者中总共鉴定出104种差异表达蛋白，并用MRM分析验证了26种潜在生物标志物。七种蛋白（BSG、HSP90AB1、SLC44A1、CHGA、ATP6V0A1、ITGAL和SELL）显示出强大的能力，能够将NB患者与健康对照以及非NB患者区分开来。scRNA-seq和sEVs蛋白质组学的综合分析表明，这些源自sEVs的生物标志物起源于肿瘤组织中的不同细胞群体。
基于sEVs的生物标志物可能有助于NB的分子诊断，代表了一种改善NB检测和管理的创新策略。
