TY - JOUR
T1 - Extracellular vesicles have variable dose-dependent effects on cultured draining cells in the eye
AU - Tabak, Saray
AU - Schreiber-Avissar, Sofia
AU - Beit-Yannai, Elie
N1 - Funding Information:
This work was supported by grants from the ISRAEL SCIENCE FOUNDATION (grant No. 1315/14). We thank Dr. Uzi Hadad from the Ilse Katz Institute for Nanoscale Science &Technology at Ben-Gurion University for his support in Image Stream analysis. Funding source: The sponsor or funding organization had no role in the research. Performed the research; S.T. Designed the research study; S.S.A. and E.B.Y. Analysed the data and wrote the paper; S.T., S.S.A. and E.B.Y.
Funding Information:
This work was supported by grants from the ISRAEL SCIENCE FOUNDATION (grant No. 1315/14). We thank Dr. Uzi Hadad from the Ilse Katz Institute for Nanoscale Science &Technology at Ben-Gurion University for his support in Image Stream analysis.
Publisher Copyright:
© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.
PY - 2018/3/1
Y1 - 2018/3/1
N2 - The role of extracellular vesicles (EVs) as signal mediators has been described in many biological fields. How many EVs are needed to deliver the desired physiological signal is yet unclear. Using a normal trabecular meshwork (NTM) cell culture exposed to non-pigmented ciliary epithelium (NPCE)–derived EVs, a relevant model for studying the human ocular drainage system, we addressed the EVs dose–response effects on the Wnt signaling. The objective of the study was to investigate the dosing effects of NPCE-derived EVs on TM Wnt signaling. EVs were isolated by PEG 8000 method from NPCE and RPE cells (used as controls) conditioned media. Concentrations were determined by Tunable Resistive Pulse Sensing method. Various exosomes concentration were incubated with TM cells, for the determination of mRNA (β-Catenin, Axin2 and LEF1) and protein (β-Catenin, GSK-3β) expression using real-time quantitative PCR and Western blot, respectively. Exposure of NTM cells for 8 hrs to low EVs concentrations was associated with a significant decreased expression of β-Catenin, GSK-3β, as opposed to exposure to high exosomal concentrations. Pro-MMP9 and MMP9 activities were significantly enhanced in NTM cells treated with high EV concentrations of (X10) as compared to low EV concentrations of either NPCE- or RPE-derived EVs and to untreated control. Our data support the concept that EVs biological effects are concentration-dependent at their target site. Specifically in the present study, we described a general dose–response at the gene and MMPs activity and a different dose–response regarding key canonical Wnt proteins expression.
AB - The role of extracellular vesicles (EVs) as signal mediators has been described in many biological fields. How many EVs are needed to deliver the desired physiological signal is yet unclear. Using a normal trabecular meshwork (NTM) cell culture exposed to non-pigmented ciliary epithelium (NPCE)–derived EVs, a relevant model for studying the human ocular drainage system, we addressed the EVs dose–response effects on the Wnt signaling. The objective of the study was to investigate the dosing effects of NPCE-derived EVs on TM Wnt signaling. EVs were isolated by PEG 8000 method from NPCE and RPE cells (used as controls) conditioned media. Concentrations were determined by Tunable Resistive Pulse Sensing method. Various exosomes concentration were incubated with TM cells, for the determination of mRNA (β-Catenin, Axin2 and LEF1) and protein (β-Catenin, GSK-3β) expression using real-time quantitative PCR and Western blot, respectively. Exposure of NTM cells for 8 hrs to low EVs concentrations was associated with a significant decreased expression of β-Catenin, GSK-3β, as opposed to exposure to high exosomal concentrations. Pro-MMP9 and MMP9 activities were significantly enhanced in NTM cells treated with high EV concentrations of (X10) as compared to low EV concentrations of either NPCE- or RPE-derived EVs and to untreated control. Our data support the concept that EVs biological effects are concentration-dependent at their target site. Specifically in the present study, we described a general dose–response at the gene and MMPs activity and a different dose–response regarding key canonical Wnt proteins expression.
KW - Exosomes
KW - Extracellular vesicles
KW - Non pigmented ciliary epithelium
KW - Signaling
KW - Trabecular meshwork
KW - Wnt
UR - http://www.scopus.com/inward/record.url?scp=85041650328&partnerID=8YFLogxK
U2 - 10.1111/jcmm.13505
DO - 10.1111/jcmm.13505
M3 - Article
AN - SCOPUS:85041650328
SN - 1582-1838
VL - 22
SP - 1992
EP - 2000
JO - Journal of Cellular and Molecular Medicine
JF - Journal of Cellular and Molecular Medicine
IS - 3
ER -