Adipocyte-derived extracellular vesicles regulate survival and function of pancreatic β cells
Iacopo Gesmundo, … , Giovanni Camussi, Riccarda Granata
Iacopo Gesmundo, … , Giovanni Camussi, Riccarda Granata
Published February 4, 2021
Citation Information: JCI Insight. 2021;6(5):e141962. https://doi.org/10.1172/jci.insight.141962.
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Research Article Metabolism

Adipocyte-derived extracellular vesicles regulate survival and function of pancreatic β cells

Abstract

Extracellular vesicles (EVs) are implicated in the crosstalk between adipocytes and other metabolic organs, and an altered biological cargo has been observed in EVs from human obese adipose tissue (AT). Yet, the role of adipocyte-derived EVs in pancreatic β cells remains to be determined. Here, we explored the effects of EVs released from adipocytes isolated from both rodents and humans and human AT explants on survival and function of pancreatic β cells and human pancreatic islets. EVs from healthy 3T3-L1 adipocytes increased survival and proliferation and promoted insulin secretion in INS-1E β cells and human pancreatic islets, both those untreated or exposed to cytokines or glucolipotoxicity, whereas EVs from inflamed adipocytes caused β cell death and dysfunction. Human lean adipocyte-derived EVs produced similar beneficial effects, whereas EVs from obese AT explants were harmful for human EndoC-βH3 β cells. We observed differential expression of miRNAs in EVs from healthy and inflamed adipocytes, as well as alteration in signaling pathways and expression of β cell genes, adipokines, and cytokines in recipient β cells. These in vitro results suggest that, depending on the physiopathological state of AT, adipocyte-derived EVs may influence β cell fate and function.

Authors

Iacopo Gesmundo, Barbara Pardini, Eleonora Gargantini, Giacomo Gamba, Giovanni Birolo, Alessandro Fanciulli, Dana Banfi, Noemi Congiusta, Enrica Favaro, Maria Chiara Deregibus, Gabriele Togliatto, Gaia Zocaro, Maria Felice Brizzi, Raul M. Luque, Justo P. Castaño, Maria Alessandra Bocchiotti, Simone Arolfo, Stefania Bruno, Rita Nano, Mario Morino, Lorenzo Piemonti, Huy Ong, Giuseppe Matullo, Juan M. Falcón-Pérez, Ezio Ghigo, Giovanni Camussi, Riccarda Granata

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Figure 2

Effect of Ad-EVs and CK-EVs on survival, proliferation, apoptosis, and function of INS-1E β cells.

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Effect of Ad-EVs and CK-EVs on survival, proliferation, apoptosis, and f...
(A) Top: Representative fluorescence microscopy micrographs showing internalization of Ad-EVs in INS-1E β cells, incubated at the indicated times with unlabeled Ad-EVs (Control) or with Ad-EVs labeled with PKH26 (red dye) (scale bar: 10 μm). Bottom: Internalization of Ad-EVs for 24 hours in cells untreated (-CD29) or preincubated for 1 hour with the blocking antibody against CD29 (+CD29). Nuclei were stained blue with DAPI (scale bar: 10 μm). Cell survival (B), cell proliferation (C), and apoptosis (D) assessed by MTT, BrdU, and caspase-3 activity, respectively, in cells cultured in serum-deprived medium for 12 hours, then untreated (control [c]) or treated for a further 24 hours with Ad-EVs (10 × 103/cell) or with EVs from 3T3-L1 adipocytes treated for 24 hours with cytokines (CKs) (CK-EVs) (TNF-α/IFN-γ/IL-1β [50, 25, and 2.5 ng/mL, respectively]). (E) Cell survival in cells treated for 24 hours with or without Ad-EVs and anti-CD29 antibody. Results are expressed as percentage of control (mean ± SEM). *P < 0.05, **P < 0.01, ***P < 0.001 vs. c; ##P < 0.01, ###P < 0.001 by 1-way ANOVA and Tukey’s post hoc test (n = 5 for B and C; n = 3 for D; n = 4 for E). Cell survival (F), cell proliferation (G), and apoptosis (H) assessed by MTT, BrdU, and caspase-3 activity, respectively, in β cells cultured in serum-deprived medium or pretreated for 40 minutes with CKs (TNF-α/IFN-γ/IL-1β [100, 50 and 5 ng/mL, respectively]), and then with EVs or CK-EVs for 24 hours. (I) Cell survival in cells cultured with or without CKs, Ad-EVs, and CD29 blocking antibody. Results are expressed as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. CK; ##P < 0.01, ###P < 0.001 by 1-way ANOVA and Tukey’s post hoc test (n = 5 for F and G; n = 3 for H; n = 4 for I). (J and K) Insulin secretion assessed by ELISA in INS-1E β cells incubated with 2 mM glucose for 1 hour and for a further 1 hour with the indicated concentrations of glucose, in the presence or absence of Ad-EVs (J) or CK-EVs (K) (mean ± SEM). *P < 0.05 by 2-tailed Student’s t test (n = 3).