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%).

Background Recent trends of hepatocellular carcinoma (HCC) mortality and outcome remain unknown in the United States (US). We investigated the recent trends of primary liver cancer (excluding intrahepatic cholangiocarcinoma) mortality and HCC stage, treatment, and overall survival (OS) in the US. Methods US Cancer Mortality database was analyzed to investigate the trend of primary liver cancer mortality. We analyzed the SEER 18 database to assess the temporal trend of tumor size, stage, treatment, and OS of HCC. Cox regression analysis investigated the association between HCC diagnosis year and OS. All statistical tests were 2-sided. Results During 2000-2018, liver cancer mortality rates increased until 2013, plateaued during 2013-2016 (annual percent change [APC] = 0.1%/yr, 95% confidence interval [CI] = -2.1% to 2.4%; P=0.92), and started to decline during 2016-2018 (APC = -1.5%/yr, 95% CI= -3.2% to 0.2%; P=0.08). However, mortality continues to increase in American Indians/Alaska Natives, individuals aged 65 or older, and in 33 states. There was a 0.61% (95% CI = 0.53% to 0.69%; P<0.001) increase in localized stage HCC and 0.86 mm (95% CI= -1.10 to -0.62; P<0.001) decrease in median tumor size per year. One-year OS rate increased from 36.3% (95% CI = 34.3% to 38.3%) to 58.1% (95% CI = 56.9% to 59.4%) during 2000-2015, and five-year OS rate almost doubled from 11.7% (95% CI = 10.4% to 13.1%) to 21.3% (95% CI = 20.2% to 22.4%) during 2000-2011. Diagnosis year (per year) (adjusted hazard ratio = 0.96; 95% CI = 0.96 to 0.97) was independently associated with OS in multivariable analysis. Conclusions Primary liver cancer mortality rates have started to decline in the US with demographic and state-level variation. With an increasing detection of localized HCC, the OS of HCC has improved over the past decades.

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and one of the leading causes of cancer-related death worldwide. Despite the improvements in surveillance and treatment, the prognosis of HCC remains poor. Extracellular vesicles (EVs) are a heterogeneous group of phospholipid bilayer-enclosed particles circulating in the bloodstream and mediating intercellular communication. Emerging studies have shown that EVs play a crucial role in regulating the proliferation, immune escape, and metastasis of HCC. In addition, because EVs are present in the circulation at relatively early stages of disease, they are getting attention as an attractive biomarker for HCC detection. Over the past decade, dedicated efforts have been made to isolate EVs more efficiently and make them useful tools in different clinical settings. In this review article, we provide an overview of the EVs isolation methods and highlight the role of EVs as mediators in the pathogenesis and progression of HCC. Lastly, we summarize the potential applications of EVs in early-stage HCC detection.