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

doi:10.1016/j.celrep.2019.05.095

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

Department of Developmental Biology and Cancer Research

Research output: Contribution to journal › Article › peer-review

155 Scopus citations

Abstract

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 language	American English
Pages (from-to)	3956-3971.e6
Journal	Cell Reports
Volume	27
Issue number	13
DOIs	https://doi.org/10.1016/j.celrep.2019.05.095
State	Published - 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)

Keywords

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

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10.1016/j.celrep.2019.05.095

Cite this

Borghesan, M., Fafián-Labora, J., Eleftheriadou, O., Carpintero-Fernández, P., Paez-Ribes, M., Vizcay-Barrena, G., Swisa, A., Kolodkin-Gal, D., Ximénez-Embún, P., Lowe, R., Martín-Martín, B., Peinado, H., Muñoz, J., Fleck, R. A., Dor, Y., Ben-Porath, I., Vossenkamper, A., Muñoz-Espin, D., & O'Loghlen, A. (2019). Small Extracellular Vesicles Are Key Regulators of Non-cell Autonomous Intercellular Communication in Senescence via the Interferon Protein IFITM3. Cell Reports, 27(13), 3956-3971.e6. https://doi.org/10.1016/j.celrep.2019.05.095

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

abstract = "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.",

keywords = "DDIS, EV, IFITM3, OIS, aging, exosomes, fragilis, interferon, paracrine senescence, small extracellular vesicles",

author = "Michela Borghesan and Juan Fafi{\'a}n-Labora and Olga Eleftheriadou and Paula Carpintero-Fern{\'a}ndez and Marta Paez-Ribes and Gema Vizcay-Barrena and Avital Swisa and Dror Kolodkin-Gal and Pilar Xim{\'e}nez-Emb{\'u}n and Robert Lowe and Belen Mart{\'i}n-Mart{\'i}n and Hector Peinado and Javier Mu{\~n}oz and Fleck, {Roland A.} and Yuval Dor and Ittai Ben-Porath and Anna Vossenkamper and Daniel Mu{\~n}oz-Espin and Ana O'Loghlen",

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.{\textquoteright}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.{\textquoteright}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: {\textcopyright} 2019 The Author(s)",

year = "2019",

month = jun,

day = "25",

doi = "10.1016/j.celrep.2019.05.095",

language = "American English",

volume = "27",

pages = "3956--3971.e6",

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Borghesan, M, Fafián-Labora, J, Eleftheriadou, O, Carpintero-Fernández, P, Paez-Ribes, M, Vizcay-Barrena, G, Swisa, A, Kolodkin-Gal, D, Ximénez-Embún, P, Lowe, R, Martín-Martín, B, Peinado, H, Muñoz, J, Fleck, RA, Dor, Y , Ben-Porath, I, Vossenkamper, A, Muñoz-Espin, D & O'Loghlen, A 2019, 'Small Extracellular Vesicles Are Key Regulators of Non-cell Autonomous Intercellular Communication in Senescence via the Interferon Protein IFITM3', Cell Reports, vol. 27, no. 13, pp. 3956-3971.e6. https://doi.org/10.1016/j.celrep.2019.05.095

TY - JOUR

T1 - Small Extracellular Vesicles Are Key Regulators of Non-cell Autonomous Intercellular Communication in Senescence via the Interferon Protein IFITM3

AU - Borghesan, Michela

AU - Fafián-Labora, Juan

AU - Eleftheriadou, Olga

AU - Carpintero-Fernández, Paula

AU - Paez-Ribes, Marta

AU - Vizcay-Barrena, Gema

AU - Swisa, Avital

AU - Kolodkin-Gal, Dror

AU - Ximénez-Embún, Pilar

AU - Lowe, Robert

AU - Martín-Martín, Belen

AU - Peinado, Hector

AU - Muñoz, Javier

AU - Fleck, Roland A.

AU - Dor, Yuval

AU - Ben-Porath, Ittai

AU - Vossenkamper, Anna

AU - Muñoz-Espin, Daniel

AU - O'Loghlen, Ana

N1 - 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)

PY - 2019/6/25

Y1 - 2019/6/25

N2 - 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.

AB - 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.

KW - DDIS

KW - EV

KW - IFITM3

KW - OIS

KW - aging

KW - exosomes

KW - fragilis

KW - interferon

KW - paracrine senescence

KW - small extracellular vesicles

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U2 - 10.1016/j.celrep.2019.05.095

DO - 10.1016/j.celrep.2019.05.095

M3 - Article

C2 - 31242426

AN - SCOPUS:85067289517

SN - 2211-1247

VL - 27

SP - 3956-3971.e6

JO - Cell Reports

JF - Cell Reports

IS - 13

ER -

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

Abstract

Bibliographical note

Keywords

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