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

• Published in: Nature communications 2020-09, Vol.11 (1), p.4489-12, Article 4489
• Main Authors: Sun, Na, Lee, Yi-Te, Zhang, Ryan Y., Kao, Rueihung, Teng, Pai-Chi, Yang, Yingying, Yang, Peng, Wang, Jasmine J., Smalley, Matthew, Chen, Pin-Jung, Kim, Minhyung, Chou, Shih-Jie, Bao, Lirong, Wang, Jing, Zhang, Xinyue, Qi, Dongping, Palomique, Juvelyn, Nissen, Nicolas, Han, Steven-Huy B., Sadeghi, Saeed, Finn, Richard S., Saab, Sammy, Busuttil, Ronald W., Markovic, Daniela, Elashoff, David, Yu, Hsiao-hua, Li, Huiying, Heaney, Anthony P., Posadas, Edwin, You, Sungyong, Yang, Ju Dong, Pei, Renjun, Agopian, Vatche G., Tseng, Hsian-Rong, Zhu, Yazhen
• Format: Article
• Language: English
• Subjects: 13/1; 13/51; 13/62; 147/135; 147/143; 38/39; 38/77; 49/90; 631/1647/350/877; 631/67/1504/1610/4029; 631/67/2322; 692/53/2421; Aged; Antibodies; Biomarkers, Tumor - genetics; Biomarkers, Tumor - isolation & purification; Cancer; Carcinoma, Hepatocellular - blood; Carcinoma, Hepatocellular - diagnosis; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Case-Control Studies; Chemistry; Click Chemistry - instrumentation; Click Chemistry - methods; Computational Chemistry; Computer Simulation; Covalence; Diagnosis, Differential; Digital signatures; Dimethylpolysiloxanes - chemistry; Disease Progression; Early Detection of Cancer - instrumentation; Early Detection of Cancer - methods; Extracellular vesicles; Extracellular Vesicles - genetics; Female; Hep G2 Cells; Hepatocellular carcinoma; Humanities and Social Sciences; Humans; Lab-On-A-Chip Devices; Liquid Biopsy - instrumentation; Liquid Biopsy - methods; Liver cancer; Liver Cirrhosis - blood; Liver Cirrhosis - pathology; Liver Neoplasms - blood; Liver Neoplasms - diagnosis; Liver Neoplasms - genetics; Liver Neoplasms - pathology; Male; Microfluidic Analytical Techniques - instrumentation; Microfluidic Analytical Techniques - methods; Microfluidics; Middle Aged; Mixers; multidisciplinary; Nanostructures - chemistry; Nanowires - chemistry; Neoplasm Staging; Purification; Reverse Transcriptase Polymerase Chain Reaction - instrumentation; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA, Messenger - genetics; RNA, Messenger - isolation & purification; ROC Curve; Science; Science (multidisciplinary); Substrates; Therapeutic applications; Vesicles
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-18311-0

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%). Extracellular vesicles (EVs) are present in circulation at relatively early stages of disease, providing potential opportunities for early cancer diagnosis. Here, the authors report a covalent chemistry-based hepatocellular carcinoma (HCC)-specific EV purification system for early detection of HCC by performing digital scoring on the purified EVs.