Signature mRNA markers in extracellular vesicles for the accurate diagnosis of colorectal cancer

Abstract

Background With the increasing incidence of colorectal cancer (CRC), its accurate diagnosis is critical and in high demand. However, conventional methods are not ideal due to invasiveness and low accuracy. Herein, we aimed to identify efficient CRC mRNA markers in a non-invasive manner using CRC-derived extracellular vesicles (EVs). The expression levels of EV mRNAs from cancer cell lines were compared with those of a normal cell line using quantitative polymerase chain reaction. Eight markers were evaluated in plasma EVs from CRC patients and healthy controls. The diagnostic value of each marker, individually or in combination, was then determined using recessive operating characteristics analyses and the Mann-Whitney U test. Results Eight mRNA markers (MYC, VEGF, CDX2, CD133, CEA, CK19, EpCAM, and CD24) were found to be more abundant in EVs derived from cancer cell lines compared to control cell lines. A combination of VEGF and CD133 showed the highest sensitivity (100%), specificity (80%), and accuracy (93%) and an area under the curve of 0.96; hence, these markers were deemed to be the CRC signature. Moreover, this signature was found to be highly expressed in CRC-derived EVs compared to healthy controls. Conclusions VEGF and CD133 mRNAs comprise a unique CRC signature in EVs that has the potential to act as a novel, non-invasive, and accurate biomarker that would improve the current diagnostic platform for CRC, while also serving to strengthen the value of EV mRNA as diagnostic markers for myriad of diseases.

Biological Functions Driven by mRNAs Carried by Extracellular Vesicles in Cancer

“…Studies in cell lines, as previously shown, have provided insight into possible biomarkers, but these results still have to be confirmed in patients to ascertain the real potential of these molecules as biomarkers. As an example, Cha et al (2020) selected a total of 12 mRNAs that were differentially expressed in EVs from 4 colorectal cancer cell lines, and through a series of comparisons, identified that the combination of only two of these mRNAs— CD133 and VEGF —was able to distinguish colorectal cancer patients from healthy controls. However, the combination of these two mRNAs achieved 100% sensitivity with extremely high specificity and accuracy.…”

The Roadmap of Colorectal Cancer Screening

“…Cha et al [ 143 ] evaluated eight mRNA markers (MYC, VEGF, CDX2, CD133, CEA, CK19, EpCAM, and CD24) extracted from plasma EVs. Of the eight mRNAs, the combination of VEGF and CD133 showed statistically significant differences between healthy and CRC, and obtained an AUC of 0.96 with 100% sensitivity and 80% specificity in discriminating between the two groups.…”

CRISPR/Cas13-Based Platforms for a Potential Next-Generation Diagnosis of Colorectal Cancer through Exosomes Micro-RNA Detection: A Review

“…Although current molecular biomarkers have a good performance, these perform correctly only within a small population of patients at specific CRC stages and with specific molecular characteristics, rendering them insufficient for a wide-range CRC diagnosis [ 20 , 94 ]. Thus, the research community keeps moving forward to develop new and more accessible molecular biomarkers that can be found more abundantly and with a broader diagnosis range, capable of diagnosing most CRC stages despite its heterogeneity, since CRC early diagnosis is critical for survival and is currently in high demand [ 95 , 96 ].…”