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Proteo-metabolomic analysis of plasma exosomes identifies novel biomarkers of thrombo-inflammation and disease progression in pediatric neuroblastoma

ABSTRACT: Neuroblastoma (NB) is a heterogeneous pediatric solid tumor in which minimally invasive biomarkers that reflect tumor biology and treatment response remain limited. Plasma exosomes contain proteins and metabolites that may provide insight into neuroblastoma pathophysiology and enable the identification of mechanistically linked biomarkers. This study employed an integrated proteo-metabolomic approach to identify novel exosomal biomarkers associated with disease status. Plasma samples were obtained from 14 patients with neuroblastoma before tumor resection (on-disease), 15 patients after tumor resection (follow-up), and 20 healthy donors. Plasma exosomes were characterized by transmission electron microscopy, nanoparticle tracking analysis, and immunoblotting. SWATH-targeted proteomic profiling identified 16 significantly altered exosomal proteins, and bioinformatic analysis revealed platelet activation, coagulation cascades, and growth-related signaling, all centralized on exosomal alpha-2-macroglobulin (A2M), which showed the highest level in the on-disease group and declined in the follow-up and healthy control groups.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Wararat Chiangjong

PROVIDER: PXD074613 | JPOST Repository | Thu Feb 19 00:00:00 GMT 2026

REPOSITORIES: jPOST

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Project description:Introduction: microRNAs are promising candidate breast cancer biomarkers due to their cancer-specific expression profiles. However, efforts to develop circulating breast cancer biomarkers are challenged by the heterogeneity of microRNAs in the blood. To overcome this challenge, we aimed to develop a molecular profile of microRNAs specifically secreted from breast cancer cells. Our first step towards this direction relates to capturing and analyzing the contents of exosomes, which are small secretory vesicles that selectively encapsulate microRNAs indicative of their cell of origin. To our knowledge, circulating exosome microRNAs have not been well evaluated as biomarkers for breast cancer diagnosis or monitoring. Methods: Exosomes were collected from the conditioned media of human breast cancer cell lines, mouse plasma of patient-derived orthotopic xenograft models (PDX), and human plasma samples. Exosomes were verified by electron microscopy, nanoparticle tracking analysis, and western blot. Cellular and exosome microRNAs from breast cancer cell lines were profiled by next-generation small RNA sequencing. Plasma exosome microRNA expression was analyzed by qRT-PCR analysis. Results: Small RNA sequencing and qRT-PCR analysis showed that several microRNAs are selectively encapsulated or highly enriched in breast cancer exosomes. Importantly, the selectively enriched exosome microRNA, human miR-1246, was detected at significantly higher levels in exosomes isolated from PDX mouse plasma, indicating that tumor exosome microRNAs are released into the circulation and can serve as plasma biomarkers for breast cancer. This observation was extended to human plasma samples where miR-1246 and miR-21 were detected at significantly higher levels in the plasma exosomes of 16 breast cancer patients as compared to the plasma exosomes of healthy control subjects. Receiver Operating Characteristic (ROC) curve analysis indicated that the combination of plasma exosome miR-1246 and miR-21 levels is a better indicator of breast cancer than their individual levels. Conclusions: Our results demonstrate that certain microRNA species, such as miR-21 and miR-1246, are selectively enriched in human breast cancer exosomes and significantly elevated in the plasma of breast cancer patients. These findings indicate a potential new strategy to selectively analyze plasma breast cancer microRNAs indicative of the presence of breast cancer.

			Action	DRS
	EV_21_SWATH2.wiff	Wiff
	EV_21_SWATH2.wiff.scan	Wiff
	EV_41_SWATH2.wiff	Wiff
	EV_41_SWATH2.wiff.scan	Wiff
	EV_48_IDA1.wiff	Wiff

Dataset Information

Proteo-metabolomic analysis of plasma exosomes identifies novel biomarkers of thrombo-inflammation and disease progression in pediatric neuroblastoma

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