Small Extracellular Vesicles Are Key Regulators of Non-cell Autonomous Intercellular Communication in Senescence via the Interferon Protein IFITM3

Michela Borghesan, Juan Fafián-Labora, Olga Eleftheriadou, Paula Carpintero-Fernández, Marta Paez-Ribes, Gema Vizcay-Barrena, Avital Swisa, Dror Kolodkin-Gal, Pilar Ximénez-Embún, Robert Lowe, Belen Martín-Martín, Hector Peinado, Javier Muñoz, Roland A. Fleck, Yuval Dor, Ittai Ben-Porath, Anna Vossenkamper, Daniel Muñoz-Espin, Ana O'Loghlen*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

155 Scopus citations


Senescence is a cellular phenotype present in health and disease, characterized by a stable cell-cycle arrest and an inflammatory response called senescence-associated secretory phenotype (SASP). The SASP is important in influencing the behavior of neighboring cells and altering the microenvironment; yet, this role has been mainly attributed to soluble factors. Here, we show that both the soluble factors and small extracellular vesicles (sEVs) are capable of transmitting paracrine senescence to nearby cells. Analysis of individual cells internalizing sEVs, using a Cre-reporter system, show a positive correlation between sEV uptake and senescence activation. We find an increase in the number of multivesicular bodies during senescence in vivo. sEV protein characterization by mass spectrometry (MS) followed by a functional siRNA screen identify interferon-induced transmembrane protein 3 (IFITM3) as being partially responsible for transmitting senescence to normal cells. We find that sEVs contribute to paracrine senescence. Borghesan et al. show that the soluble fraction and small extracellular vesicles (sEVs) mediate paracrine senescence. RNA sequencing and loxP reporter systems confirm sEV-mediated paracrine senescence, while preventing sEV release averts senescence. Mass spectrometry and functional analysis show that the IFN protein, IFITM3, is partially responsible for this phenotype.

Original languageAmerican English
Pages (from-to)3956-3971.e6
JournalCell Reports
Issue number13
StatePublished - 25 Jun 2019

Bibliographical note

Funding Information:
We are grateful to Tom Nightingale and Maria Niklison-Chirou for reading the manuscript. Alissa Weaver provided tagged CD63 constructs; and Jacob Yount and I-Chueh Huang supplied the IFITM3 and shIFITM3 plasmids. We are grateful to Luke Gammon, the Queen Mary University of London (QMUL) Genome Centre, and Gary Warnes for excellent technical support. Mouse hepatic stellate cells were a gift from Scott Lowe. A.O.’s lab is supported by the BBSRC (BB/P000223/1) and The Royal Society (RG170399). M.B. is funded by the MRC (MR/K501372/1) and the Centre for Genomics and Child Health. P.C.-F. (IN606B 2017/014) and J.F.-L. (ED481B 2017/117) are funded by the Xunta de Galicia. M.B. and J.F.-L. performed most of the experiments, with help from O.E. and P.C.-F. M.P.-R. and D.M.-E. performed the IHC staining and analysis. G.V.-B. and R.A.F. performed the TEM. A.S. D.K.-G. Y.D. and I.B.-P. provided the fixed mouse pancreatic tissue. P.X.-E. and J.M. performed the proteomics analyses. R.L. performed the RNA-seq analysis. B.M.-M. helped with the imaging analysis. H.P. provided advice regarding the proteomic experimental settings. A.V. performed the blood donor experiments. A.O. conceived and designed the study and wrote and edited the manuscript, with input from all of the authors. The authors declare no competing interests.

Funding Information:
A.O.’s lab is supported by the BBSRC ( BB/P000223/1 ) and The Royal Society ( RG170399 ). M.B. is funded by the MRC ( MR/K501372/1 ) and the Centre for Genomics and Child Health . P.C.-F. ( IN606B 2017/014 ) and J.F.-L. ( ED481B 2017/117 ) are funded by the Xunta de Galicia .

Publisher Copyright:
© 2019 The Author(s)


  • DDIS
  • EV
  • IFITM3
  • OIS
  • aging
  • exosomes
  • fragilis
  • interferon
  • paracrine senescence
  • small extracellular vesicles


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