miR-18a and miR-106a Signatures in Plasma Small EVs Are Promising Biomarkers for Early Detection of Pancreatic Ductal Adenocarcinoma

Abstract

Pancreatic cancer is the third leading cause of cancer-related death in the United States. Pancreatic ductal adenocarcinoma (PDAC) is the major form of pancreatic cancer with the worst outcomes. Early detection is key to improving the overall survival rate of PDAC patients. Recent studies have demonstrated that microRNA (miRNA) signatures in plasma small extracellular vesicles (EVs) are potential biomarkers for the early detection of PDAC. However, published results are inconsistent due to the heterogeneity of plasma small EVs and the methods used for small EV isolation. We have recently refined the process of plasma small EV isolation using double filtration and ultracentrifugation. In the present study, we applied this protocol and analyzed plasma small EV miRNA signatures by small RNA sequencing and quantitative RT-PCR in a pilot cohort, consisting of patients with early-stage PDAC, and age- and gender-matched healthy subjects (n = 20). We found, via small RNA sequencing, that there are several miRNAs enriched in plasma small EVs of PDAC patients, and the levels of miR-18a and miR-106a were confirmed by quantitative RT-PCR to be significantly elevated in patients with early-stage PDAC compared with age- and gender-matched healthy subjects. Furthermore, using an immunoaffinity-based plasma small EV isolation approach, we confirmed that the levels of miR-18a and miR-106a in plasma small EVs were significantly higher in PDAC patients versus the healthy subjects. We thus conclude that the levels of miR-18a and miR-106a in plasma small EVs are promising biomarkers for the early detection of PDAC.

Keywords: biomarkers; early detection; extracellular vesicles; microRNAs; pancreatic cancer.

Figure 1

Elevated expression of selected miRNAs in small EVs isolated from PDAC patient plasma. Small EVs were isolated from plasma (250 μL each) of patients with early-stage PDAC and age- and gender-matched healthy subjects (HP). miRNA expression was analyzed by qRT-PCR. Expression of miR-1246 ((A), n = 10), miR-106a ((B), n = 19), and miR-18a ((C), n = 20) is shown. ** p < 0.01, paired Student’s t-test.