Open Access

Bogdan Mateescu, Emma Kowal, Bastiaan Wilhelmus Maria van Balkom, Sabine Bartel, Suvendra N. Bhattacharyya, Edit I. Buzás, Amy H. Buck, Paola de-Candia, Franklin W. N. Chow, Saumya Das, Tom A. P. Driedonks, Lola Fernández-Messina, Franziska Haderk, Andrew F. Hill, Jennifer C. Jones, Kendall R. Van Keuren-Jensen, Charles P. Lai, Cecilia Lässer, Italia Di Liegro, Taral Rameshchand Lunavat, Magdalena J. Lorenowicz, Sybren L. N. Maas, Imre Mäger, Maria Mittelbrunn, Stefan Momma, Kamalika Mukherjee, Kamalika Mukherjee, Muhammed Nawaz, Dirk Michiel Pegtel, Michael W. Pfaff, Raymond M. Schiffelers, Hidetoshi Tahara, Clotilde Théry, Juan Pablo Tosar, Marca H. M. Wauben, Kenneth W. Witwer, Esther N. M. Nolte-’t Hoen

• The release of RNA-containing extracellular vesicles (EV) into the extracellular milieu has been demonstrated in a multitude of different in vitro cell systems and in a variety of body fluids. RNA-containing EV are in the limelight for their capacity to communicate genetically encoded messages to other cells, their suitability as candidate biomarkers for diseases, and their use as therapeutic agents. Although EV-RNA has attracted enormous interest from basic researchers, clinicians, and industry, we currently have limited knowledge on which mechanisms drive and regulate RNA incorporation into EV and on how RNA-encoded messages affect signalling processes in EV-targeted cells. Moreover, EV-RNA research faces various technical challenges, such as standardisation of EV isolation methods, optimisation of methodologies to isolate and characterise minute quantities of RNA found in EV, and development of approaches to demonstrate functional transfer of EV-RNA in vivo. These topics were discussed at the 2015 EV-RNA workshop of the International Society for Extracellular Vesicles. This position paper was written by the participants of the workshop not only to give an overview of the current state of knowledge in the field, but also to clarify that our incomplete knowledge – of the nature of EV(-RNA)s and of how to effectively and reliably study them – currently prohibits the implementation of gold standards in EV-RNA research. In addition, this paper creates awareness of possibilities and limitations of currently used strategies to investigate EV-RNA and calls for caution in interpretation of the obtained data.