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Published on: February 27, 2015
Quantitative proteomics identifies and validates urinary biomarkers of rhabdomyosarcoma in children
Na Xu1,2, Yuncui Yu3, Chao Duan1
1Medical 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.
Insights
This study identifies novel urinary biomarkers for early rhabdomyosarcoma (RMS) detection. These findings enable noninvasive diagnosis and improve outcomes for pediatric cancer patients.
Area of Science:
- Biochemistry
- Oncology
- Proteomics
Background:
- Rhabdomyosarcoma (RMS) is a prevalent childhood soft tissue sarcoma with a poor prognosis in advanced stages.
- Early detection through noninvasive methods like urine analysis is crucial for improving treatment outcomes.
- Urine proteomics offers a promising avenue for identifying low-abundance tumor markers.
Purpose of the Study:
- To identify and validate novel urinary protein biomarkers for the early, noninvasive diagnosis of rhabdomyosarcoma (RMS).
- To establish a diagnostic panel for improved screening of pediatric RMS patients.
Main Methods:
- A two-stage proteomic workflow was employed, involving discovery and verification phases using mass spectrometry-based methods (data-independent acquisition and parallel reaction monitoring).
- Urine samples from pediatric RMS patients and healthy controls were analyzed.
- Bioinformatics tools, including Gene Ontology and Ingenuity Pathway Analysis, were utilized for data interpretation.
Main Results:
- A total of 251 significantly altered proteins were identified in the discovery stage, with enrichment in common RMS sites.
- 39 proteins were confirmed as potential urinary biomarkers for RMS.
- A diagnostic panel of 5 proteins (EPS8L2, SPARC, HLA-DRB1, ACAN, CILP) demonstrated diagnostic potential with an AUC of 0.79.
Conclusions:
- Novel urinary biomarkers for RMS have been identified, facilitating easier clinical translation for noninvasive molecular diagnosis.
- Urine proteomics is valuable for identifying and qualifying candidate biomarkers for early cancer detection.
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.
