定量蛋白质组学鉴定并验证了儿童横纹肌肉瘤的尿液生物标志物。

Quantitative proteomics identifies and validates urinary biomarkers of rhabdomyosarcoma in children.

作者信息

Xu Na, Yu Yuncui, Duan Chao, Wei Jing, Sun Wei, Jiang Chiyi, Jian Binglin, Cao Wang, Jia Lulu, Ma Xiaoli

机构信息

Medical Oncology Department, Pediatric Oncology Center, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing Key Laboratory of Pediatric Hematology Oncology, Key Laboratory of Major Diseases in Children, Ministry of Education, No. 56 Nalishi Road, Beijing, 100045, China.

Department of Pediatrics, Beijing Friendship Hospital, Capital Medical University, Beijing, China.

出版信息

Clin Proteomics. 2023 Mar 14;20(1):10. doi: 10.1186/s12014-023-09401-4.

DOI:10.1186/s12014-023-09401-4

PMID:36918772

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10012572/

Abstract

BACKGROUND

Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma with poor prognosis in children. The 5-year survival rate for early RMS has improved, whereas it remains unsatisfactory for advanced patients. Urine can rapidly reflect changes in the body and identify low-abundance proteins. Early screening of tumor markers through urine in RMS allows for earlier treatment, which is associated with better outcomes.

METHODS

RMS patients under 18 years old, including those newly diagnosed and after surgery, were enrolled. Urine samples were collected at the time points of admission and after four cycles of chemotherapy during follow-up. Then, a two-stage workflow was established. (1) In the discovery stage, differential proteins (DPs) were initially identified in 43 RMS patients and 12 healthy controls (HCs) using a data-independent acquisition method. (2) In the verification stage, DPs were further verified as biomarkers in 54 RMS patients and 25 HCs using parallel reaction monitoring analysis. Furthermore, a receiver operating characteristic (ROC) curve was used to construct the protein panels for the diagnosis of RMS. Gene Ontology (GO) and Ingenuity Pathway Analysis (IPA) software were used to perform bioinformatics analysis.

RESULTS

A total of 251 proteins were significantly altered in the discovery stage, most of which were enriched in the head, neck and urogenital tract, consistent with the most common sites of RMS. The most overrepresented biological processes from GO analysis included immunity, inflammation, tumor invasion and neuronal damage. Pathways engaging the identified proteins revealed 33 common pathways, including WNT/β-catenin signaling and PI3K/AKT signaling. Finally, 39 proteins were confirmed as urinary biomarkers for RMS, and a diagnostic panel composed of 5 candidate proteins (EPS8L2, SPARC, HLA-DRB1, ACAN, and CILP) was constructed for the early screening of RMS (AUC: 0.79, 95%CI = 0.66 ~ 0.92).

CONCLUSIONS

These findings provide novel biomarkers in urine that are easy to translate into clinical diagnosis of RMS and illustrate the value of global and targeted urine proteomics to identify and qualify candidate biomarkers for noninvasive molecular diagnosis.

摘要

背景

横纹肌肉瘤（RMS）是儿童中最常见且预后较差的软组织肉瘤。早期RMS的5年生存率有所提高，而晚期患者的生存率仍不尽人意。尿液能够快速反映身体变化并识别低丰度蛋白质。通过尿液对RMS进行肿瘤标志物的早期筛查可实现更早治疗，这与更好的治疗结果相关。

方法

纳入18岁以下的RMS患者，包括新诊断患者及术后患者。在入院时及随访期间化疗四个周期后采集尿液样本。然后，建立了两阶段工作流程。（1）在发现阶段，最初使用数据非依赖采集方法在43例RMS患者和12例健康对照（HC）中鉴定差异蛋白（DP）。（2）在验证阶段，使用平行反应监测分析在54例RMS患者和25例HC中进一步验证DP作为生物标志物。此外，使用受试者工作特征（ROC）曲线构建用于RMS诊断的蛋白质组。使用基因本体论（GO）和 Ingenuity通路分析（IPA）软件进行生物信息学分析。

结果

在发现阶段共有251种蛋白质发生显著变化，其中大多数在头颈部和泌尿生殖道富集，这与RMS最常见的部位一致。GO分析中最具代表性的生物学过程包括免疫、炎症、肿瘤侵袭和神经元损伤。涉及已鉴定蛋白质的通路揭示了33条常见通路，包括WNT/β-连环蛋白信号通路和PI3K/AKT信号通路。最后，39种蛋白质被确认为RMS的尿液生物标志物，并构建了由5种候选蛋白质（EPS8L2、SPARC、HLA-DRB1、ACAN和CILP）组成的诊断组用于RMS的早期筛查（AUC：0.79，95%CI = 0.66 ~ 0.92）。