Purification of HCC-specific extracellular vesicles on nanosubstrates for early HCC detection by digital scoring

Abstract

We report a covalent chemistry-based hepatocellular carcinoma (HCC)-specific extracellular vesicle (EV) purification system for early detection of HCC by performing digital scoring on the purified EVs. Earlier detection of HCC creates more opportunities for curative therapeutic interventions. EVs are present in circulation at relatively early stages of disease, providing potential opportunities for HCC early detection. We develop an HCC EV purification system (i.e., EV Click Chips) by synergistically integrating covalent chemistry-mediated EV capture/release, multimarker antibody cocktails, nanostructured substrates, and microfluidic chaotic mixers. We then explore the translational potential of EV Click Chips using 158 plasma samples of HCC patients and control cohorts. The purified HCC EVs are subjected to reverse-transcription droplet digital PCR for quantification of 10 HCC-specific mRNA markers and computation of digital scoring. The HCC EV-derived molecular signatures exhibit great potential for noninvasive early detection of HCC from at-risk cirrhotic patients with an area under receiver operator characteristic curve of 0.93 (95% CI, 0.86 to 1.00; sensitivity = 94.4%, specificity = 88.5%).

Could protein content of Urinary Extracellular Vesicles be useful to detect Cirrhosis in Alcoholic Liver Disease?

“…Other authors also described changes when ALD in circulating EVs concentration 27 , 35 - 37 , certain miRNAs 27 , 37 , 95 and hepatic proteins such as ALB, HP, FGB, CYP2E1, 2A, 1A1/2, 4B 35 , 36 . For HCC diagnosis, new EVs-based approaches that allow easy and standardized use in clinical settings are being studied 96 . The proteins detected to be differently regulated in cirrhotic patients in our study are not specifically expressed in liver.…”

Could Protein Content of Urinary Extracellular Vesicles be Useful to Detect Alcoholic Liver Disease and Assess Hepatocarcinoma Risk?

“…Other authors also described changes when ALD in circulating EVs concentration [27,[61][62][63], certain miRNAs [27,63,118] and hepatic proteins such as ALB, HP, FGB, CYP2E1, 2A, 1A1/2, 4B [61,62]. For HCC diagnosis, new EVs-based approaches that allow easy and standardized use in clinical settings are being studied [119].…”

Small Extracellular Vesicles: A Novel Avenue for Cancer Management

“…Ten HCC-related mRNAs in sEVs were identified by EV Click Chip analysis and these mRNAs could be used to accurately detect early-stage HCC ( 128 ). A scoring system based on these mRNAs was superior in distinguishing HCC patients from liver cirrhosis patients.…”

“…A scoring system based on these mRNAs was superior in distinguishing HCC patients from liver cirrhosis patients. Compared with traditional serum AFP tests, this system provided a higher area under the curve (AUC) of 0.69 versus 0.93, 0.68 versus 0.91, and 0.70 versus 0.92, according to BCLC stage 0-A, Milan criteria, and UNOS DS criteria respectively ( 128 ). These data provide strong evidence of sEV mRNA utilization for early-stage cancer detection.…”