Shared extracellular vesicle miRNA profiles of matched ductal pancreatic adenocarcinoma organoids and blood plasma samples show the power of organoid technology

Abstract

Extracellular vesicles (EV) are considered as a promising diagnostic tool for pancreatic ductal adenocarcinoma (PDAC), a disease with a poor 5-year survival that has not improved in the past years. PDAC patient-derived 3D organoids maintain the intratumoral cellular heterogeneity, characteristic for the tumor in vivo.Thus, they represent an ideal in vitro model system to study human cancers. Here we show that the miRNA cargo of EVs from PDAC organoids largely differs among patients. However, we detected a common set of EV miRNAs that were present in matched organoids and blood plasma samples of individual patients. Importantly, the levels of EV miR-21 and miR-195 were higher in PDAC blood EV preparations than in healthy controls, albeit we found no difference compared to chronic pancreatitis (CP) samples. In addition, here we report that the accumulation of collagen I, a characteristic change in the extracellular matrix (ECM) in both CP and PDAC, largely increases EV release from pancreatic ductal organoids. This provides a possible explanation why both CP and PDAC patient-derived plasma samples have an elevated amount of CD63 + EVs. Collectively, we show that PDAC patient-derived organoids represent a highly relevant model to analyze the cargo of tumor cell-derived EVs. Furthermore, we provide evidence that not only driver mutations, but also changes in the ECM may critically modify EV release from pancreatic ductal cells.

The Cellular and Biological Impact of Extracellular Vesicles in Pancreatic Cancer

“…Although these studies identified such miRNAs as potential biomarkers, the cellular origins of these miRNA-carrying EVs remain unknown. A recent study from Zeöld et al confirmed this miRNA profiling by establishing shared extracellular vesicles’ miRNA profiles of matched PDA organoids and patient blood plasma samples, suggesting PDA tumor cells as cells of origin [ 24 ]. Specifically designed studies of tumor cell-derived EVs further reported the prognostic value of the tumor-suppressive miRNA, miR-192-5p [ 25 ].…”

Dysregulation of miRNAs Targeting the IGF-1R Pathway in Pancreatic Ductal Adenocarcinoma

“…A few recent studies have reported that specific long noncoding RNAs drive tumorigenesis by modulating the miR-195-5p/Wnt/beta-catenin signaling pathway [51] and promote the progression of pancreatic cancer by upregulating programmed death-ligand 1 by acting as a sponge for microRNA-195-5p [52]. Higher levels of miR-195 have also recently been reported in extracellular vesicles from blood from patients with PDAC than from blood from healthy controls, although no difference was detected when compared to extracellular vesicles from blood from patients with chronic pancreatitis [53].…”

The Past, the Present, and the Future of the Size Exclusion Chromatography in Extracellular Vesicles Separation

“…On the other hand, by separating EVs from the High Density Lipoprotein (HDL) fractions using Superose 6 or Sephacryl S-200 FPLC columns, Vickers et al showed that plasma miRNA are mostly HDL-associated [84]. In contrast, there exist studies demonstrating that extracellular RNAs are carried in EVs from human [85,86] and mouse [87] blood plasma. However, these studies did not use SEC for EV separation, making it difficult to compare.…”