Abstract
Extracellular vesicles (EVs) are found in all biological fluids, providing potential for the identification of disease biomarkers such as colorectal cancer (CRC). EVs are heavily glycosylated with specific glycoconjugates such as tetraspanins, integrins, and mucins, reflecting the characteristics of the original cell offering valuable targets for detection of CRC. We report here on europium-nanoparticle (EuNP)-based assay to detect and characterize different surface glycoconjugates of EVs without extensive purification steps from five different CRC and the HEK 293 cell lines. The promising EVs candidates from cell culture were clinically evaluated on small panel of serum samples including early-stage (n = 11) and late-stage (n = 11) CRC patients, benign condition (n = 11), and healthy control (n = 10). The majority of CRC cell lines expressed tetraspanin sub-population and glycovariants of integrins and conventional tumor markers. The subpopulation of CD151 having CD63 expression (CD151CD63) was significantly (p = 0.001) elevated in early-stage CRC (8 out of 11) without detecting any benign and late-stage samples, while conventional CEA detected mostly late-stage CRC (p = 0.045) and with only four early-stage cases. The other glycovariant assays such as CEACon-A, CA125WGA, CA 19.9Ma696, and CA 19.9Con-A further provided some complementation to the CD151CD63 assay. These results indicate the potential application of CD151CD63 assay for early detection of CRC patients in human serum.
Highlights
Diagnosing colorectal cancer (CRC) at an early stage is the key to reduce mortality and treating early-stage lesions
The CD151CD63-negative samples were further complemented with the glycovariant assays CEACon-A, CA19.9Con-A, and CA125WGA along with Carcinoembryonic antigen (CEA) EIA to detect all the early-stage (10 out of 11) and late-stage (8 out of 11) CRC serum samples
CD151CD63 assay detects only early-stage CRC; this novel assay in combination with other glycovariant (CEACon-A, CA125WGA, and CA 19.9Con-A) could be used for follow-up of CRC patients post-treatment for detection of early relapse as well as screening of CRC
Summary
Diagnosing colorectal cancer (CRC) at an early stage is the key to reduce mortality and treating early-stage lesions. The discovered promising candidates were validated with serum samples of CRC, including early and late stages; the cancer samples were tested side by side with benign disease and healthy controls and compared with conventional CEA, CA19-9, and CA125 immunoassays. The CD151CD63-negative samples were further complemented with the glycovariant assays CEACon-A, CA19.9Con-A, and CA125WGA along with CEA EIA to detect all the early-stage (10 out of 11) and late-stage (8 out of 11) CRC serum samples. Miki et al and Lewitowicsz et al reported the positive correlation of CD63 higher expression with the poor survival rate of gastric and gastrointestinal stromal tumor [39,40] In relation to these studies, we have identified the CD63 expression on CD151 expression in serum of early-stage CRC patients. CD151CD63 assay detects only early-stage CRC; this novel assay in combination with other glycovariant (CEACon-A, CA125WGA, and CA 19.9Con-A) could be used for follow-up of CRC patients post-treatment for detection of early relapse as well as screening of CRC
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