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

Anikó Zeöld,Edit I Buzás,András Áron Soós,Zoltán Wiener,Anna Kiss,Tamás Tölgyes,Ákos Szűcs,Attila Bursics,Ágnes Kittel,László Harsányi,Gyöngyvér Orsolya Sándor,András Gézsi

doi:10.1007/s00018-020-03703-8

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.

Highlights

Pancreatic ductal adenocarcinoma (PDAC) is one of the most dangerous cancers with a 5-year survival rate of less than 8%
The Extracellular vesicles (EV) concentration and miRNA profile, such as miR-21 and miR195 levels differed between PDAC plasma EV and control samples; we found no change when comparing to chronic pancreatitis (CP) patients
To develop a model system for studying PDAC-derived EV cargo, we set up 3D culture of cell lines in Matrigel, an often used extracellular matrix that is enriched for laminin and collagen type IV

Summary

Introduction

Pancreatic ductal adenocarcinoma (PDAC) is one of the most dangerous cancers with a 5-year survival rate of less than 8%. Novel promising targets have been identified [1], there is only a slow progress in the early diagnosis and treatment efficiency of this disease. Most PDAC tumors prevalently harbour mutations in KRAS, encoding a. Key protein in the signalling pathway of epidermal growth factor (EGF), and/or TP53 that is central in DNA damage responses. A large number of oncogenic driver mutations in several other genes lead to large intratumoral genetic heterogeneity [2]. Reliable biomarkers for PDAC still need to be identified. MiRNAs, among them several with known roles in cancer initiation and progression, are promising candidates. Identifying the cancer-specific expression of circulating miRNAs is obscured by the inherent heterogeneity of miRNA populations in the blood

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Results

Conclusion