MicroRNAs from Liquid Biopsy Derived Extracellular Vesicles: Recent Advances in Detection and Characterization Methods

Abstract

Liquid biopsies have become a convenient tool in cancer diagnostics, real-time disease monitoring, and evaluation of residual disease. Yet, the information still encrypted in the variety of tumor-derived molecules identified in biofluids has proven difficult to decipher due to the technological limitations imposed by their biological nature. Such is the case of extracellular vesicle (EV) encapsulated ncRNAs, which have gained traction in recent years as biomarkers. Due to their resilience towards degrading factors they may act as suitable disease indicators. This review addresses the less described issues in this context. We present an overview of less investigated biofluids that can be used for EV isolation in addition to different isolation approaches to overcome the technical challenges these specimens harbor. Furthermore, we summarize the latest technological advances providing improvement to ncRNA detection and analysis. Thereby, this review summarizes the current state-of-the-art methodologies regarding EV and EV derived miRNA analysis and how they compare to current approaches.

Liquid Biopsy in Pancreatic Cancer: Are We Ready to Apply It in the Clinical Practice?

“…Pre-analytical and analytical aspects should be considered since physiological processes such as blood hemolysis of blood during plasma collection can influence the measurement of cfRNAs [ 60 , 61 ]. The methods used for the isolation of exosomes also can also affect the quality of exosomal miRNA profiles [ 51 ].…”

“…Methods for the evaluation of cfRNAs include sequencing and RT-qPCR. The main problem in cfRNA isolation from exosomes is their low concentration, which can be addressed with more sensitive techniques such as ddPCR [ 51 ]. Recently, PCR-free techniques have been developed such as electrochemical approaches [ 64 ] or thermophoretic sensors implemented with nanoflares for in situ detection of exosomal miRNAs [ 51 , 65 ].…”

Updates on Clinical Use of Liquid Biopsy in Colorectal Cancer Screening, Diagnosis, Follow-Up, and Treatment Guidance

“…(11) The use of exosomes and CTCs in routine diagnostics is limited; only CTCs that maintain epithelial features can be detected by the EpCAM, excluding CTCs with other characteristics (Vacante et al, 2020). Regarding exosomes, one of the main challenges limiting the clinical use of these EV is the lack of standardization and consistency regarding their isolation methods, which requires difficult and laborious methods (Drula et al, 2020). In addition, the high cost of the equipment and reagents intended for the most used technology in bioscience, NGS hinders the routine use of liquid biopsy in the clinical laboratory.…”

Loss of miR-23a cluster in skeletal muscle can suppress bone remodeling

“…Among them, several miRNAs located adjacently as miRNA clusters, and dysregulation of these clusters has been identified to be involved in multiple pathogenesis events, such as, cancer, heart disease, and metabolic disorders (2). Accumulating evidence indicates that miRNAs also act as mediators of intercellular communications (3).…”