Obstacles and opportunities in the functional analysis of extracellular vesicle RNA - an ISEV position paper.
The release of RNA-containing extracellular vesicles (EV) into the extracellular milieu has been demonstrated in a multitude of differentin vitrocell 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-RNAin 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.
Department of Biology, Swiss Federal Institute of Technology Zurich (ETH Zürich) , Zurich , Switzerland.
Bogdan
B
Mateescu
Department of Biology, Massachusetts Institute of Technology , Cambridge , MA , USA.
Emma J K
0000-0002-8992-7608
EJ
Kowal
Department of Nephrology and Hypertension, UMC Utrecht , Utrecht , the Netherlands.
Bas W M
BW
van Balkom
Experimental Asthma Research, Priority Area Asthma & Allergy, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL) , Borstel , Germany.
Sabine
S
Bartel
Department of Science and Technology, CSIR-Indian Institute of Chemical Biology , Kolkata , India.
Suvendra N
SN
Bhattacharyya
Department of Genetics, Cell- and Immunobiology, Semmelweis University , Budapest , Hungary.
Edit I
EI
Buzás
Institute of Immunology and Infection Research, Centre for Immunity, Infection and Evolution, School of Biological Sciences, University of Edinburgh , Edinburgh , UK.
Amy H
AH
Buck
IRCCS MultiMedica , Milan , Italy.
Paola
P
de Candia
Institute of Immunology and Infection Research, Centre for Immunity, Infection and Evolution, School of Biological Sciences, University of Edinburgh , Edinburgh , UK.
Franklin W N
FW
Chow
Cardiovascular Research Institute, Massachusetts General Hospital , Boston , MA , USA.
Saumya
S
Das
Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University , Utrecht , the Netherlands.
Tom A P
TA
Driedonks
Immunology Service, Hospital La Princesa , Madrid , Spain.
Lola
L
Fernández-Messina
Department of Molecular Genetics, German Cancer Research Center (DKFZ), Heidelberg, Germany; Department of Medicine, Helen Diller Family Comprehensive Cancer Center, UC San Francisco, San Francisco, CA, USA.
Franziska
F
Haderk
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University , Bundoora , Australia.
Andrew F
0000-0001-5581-2354
AF
Hill
Molecular Immunogenetics & Vaccine Research Section, Vaccine Branch, CCR, NCI , Bethesda , MD , USA.
Jennifer C
0000-0002-9488-7719
JC
Jones
Neurogenomics Division, TGen , Phoenix , AZ , USA.
Kendall R
KR
Van Keuren-Jensen
Institute of Biomedical Engineering, National Tsing Hua University , Hsinchu , Taiwan.
Charles P
CP
Lai
Department of Neurology and Center for Molecular Imaging Research, Department of Radiology, Massachusetts General Hospital and NeuroDiscovery Center, Harvard Medical School, Boston, MA, USA; Krefting Research Centre, Department of Internal Medicine and Clinical Nutrition, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Cecilia
C
Lässer
Department of Experimental Biomedicine and Clinical Neurosciences (BIONEC), University of Palermo , Palermo , Italy.
Italia di
ID
Liegro
Department of Neurology and Center for Molecular Imaging Research, Department of Radiology, Massachusetts General Hospital and NeuroDiscovery Center, Harvard Medical School, Boston, MA, USA; Krefting Research Centre, Department of Internal Medicine and Clinical Nutrition, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Taral R
TR
Lunavat
Center for Molecular Medicine, University Medical Center Utrecht & Regenerative Medicine Center , Utrecht , the Netherlands.
Magdalena J
MJ
Lorenowicz
Department of Neurology and Center for Molecular Imaging Research, Department of Radiology, Massachusetts General Hospital and NeuroDiscovery Center, Harvard Medical School , Boston , MA , USA.
Sybren L N
SL
Maas
Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK; Institute of Technology, University of Tartu, Tartu, Estonia.
Imre
I
Mäger
Instituto de Investigación del Hospital 12 de Octubre , Madrid , Spain.
Maria
M
Mittelbrunn
Institute of Neurology (Edinger Institute), Frankfurt University Medical School , Frankfurt am Main , Germany.
Stefan
S
Momma
Department of Science and Technology, CSIR-Indian Institute of Chemical Biology , Kolkata , India.
Kamalika
K
Mukherjee
Department of Pathology and Forensic Medicine, Ribeirão Preto School of Medicine, University of Sao Paulo , Sao Paulo , Brazil.
Muhammed
M
Nawaz
Department of Pathology, Exosomes Research Group, VU University Medical Center , Amsterdam , the Netherlands.
D Michiel
DM
Pegtel
Animal Physiology and Immunology, School of Life Sciences, Technical University of Munich (TUM) Weihenstephan , Freising , Germany.
Michael W
MW
Pfaffl
Laboratory Clinical Chemistry & Haematology, University Medical Center Utrecht , Utrecht , the Netherlands.
Raymond M
RM
Schiffelers
Department of Cellular and Molecular Biology, Institute of Biomedical & Health Sciences, Hiroshima University , Hiroshima , Japan.
Hidetoshi
H
Tahara
Institut Curie, PSL Research University, INSERM U932 , Paris , France.
Clotilde
C
Théry
Functional Genomics Unit, Institut Pasteur de Montevideo, Nuclear Research Center, Faculty of Science, Universidad de la República , Montevideo , Uruguay.
Juan Pablo
JP
Tosar
Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University , Utrecht , the Netherlands.
Marca H M
MH
Wauben
Department of Molecular and Comparative Pathobiology and Department of Neurology, The Johns Hopkins University School of Medicine , Baltimore , MD , USA.
Kenneth W
0000-0003-1664-4233
KW
Witwer
Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University , Utrecht , the Netherlands.
Esther N M
EN
Nolte-'t Hoen
Extracellular vesicles; RNA binding proteins; exosomes; function; mRNA; non-coding RNA; quantification; sorting