TY - JOUR
T1 - Activated human astrocyte-derived extracellular vesicles modulate neuronal uptake, differentiation and firing
AU - You, Yang
AU - Borgmann, Kathleen
AU - Edara, Venkata Viswanadh
AU - Stacy, Satomi
AU - Ghorpade, Anuja
AU - Ikezu, Tsuneya
N1 - Funding Information:
This work is in part funded by Cure Alzheimer’s Fund (TI), BrightFocus Foundation (A2016551S), NIH RF1AG054199 (TI), NIH R01AG054672 (TI), NIH R56AG057469 (TI), NIH R21 NS104609 (TI), The Laboratory of Developmental Biology for provided human brain tissues as was support by NIH 5R24 HD0008836 from the Eunice Kennedy Shriver National Institute of Child Health & Human Development. We thank the Ikezu, Borgmann and Ghorpade lab members for experimental supports.
Publisher Copyright:
© 2019, © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles.
PY - 2020/1/1
Y1 - 2020/1/1
N2 - Astrocytes in the central nervous system (CNS) provide supportive neural functions and mediate inflammatory responses from microglia. Increasing evidence supports their critical roles in regulating brain homoeostasis in response to pro-inflammatory factors such as cytokines and pathogen/damage-associated molecular pattern molecules in infectious and neurodegenerative diseases. However, the underlying mechanisms of the trans-cellular communication are still unclear. Extracellular vesicles (EVs) can transfer a large diversity of molecules such as lipids, nucleic acids and proteins for cellular communications. The purpose of this study is to characterize the EVs cargo proteins derived from human primary astrocytes (ADEVs) under both physiological and pathophysiological conditions. ADEVs were isolated from human primary astrocytes after vehicle (CTL) or interleukin-1β (IL-1β) pre-treatment. Label-free quantitative proteomic profiling revealed a notable up-regulation of proteins including actin-associated molecules, integrins and major histocompatibility complex in IL-1β-ADEVs compared to CTL-ADEVs, which were involved in cellular metabolism and organization, cellular communication and inflammatory response. When fluorescently labelled ADEVs were added into primary cultured mouse cortical neurons, we found a significantly increased neuronal uptake of IL-1β-ADEVs compared to CTL-ADEVs. We further confirmed it is likely due to the enrichment of surface proteins in IL-1β-ADEVs, as IL-1β-ADEVs uptake by neurons was partially suppressed by a specific integrin inhibitor. Additionally, treatment of neurons with IL-1β-ADEVs also reduced neurite outgrowth, branching and neuronal firing. These findings provide insight for the molecular mechanism of the ADEVs’ effects on neural uptake, neural differentiation and maturation, and its alteration in inflammatory conditions.
AB - Astrocytes in the central nervous system (CNS) provide supportive neural functions and mediate inflammatory responses from microglia. Increasing evidence supports their critical roles in regulating brain homoeostasis in response to pro-inflammatory factors such as cytokines and pathogen/damage-associated molecular pattern molecules in infectious and neurodegenerative diseases. However, the underlying mechanisms of the trans-cellular communication are still unclear. Extracellular vesicles (EVs) can transfer a large diversity of molecules such as lipids, nucleic acids and proteins for cellular communications. The purpose of this study is to characterize the EVs cargo proteins derived from human primary astrocytes (ADEVs) under both physiological and pathophysiological conditions. ADEVs were isolated from human primary astrocytes after vehicle (CTL) or interleukin-1β (IL-1β) pre-treatment. Label-free quantitative proteomic profiling revealed a notable up-regulation of proteins including actin-associated molecules, integrins and major histocompatibility complex in IL-1β-ADEVs compared to CTL-ADEVs, which were involved in cellular metabolism and organization, cellular communication and inflammatory response. When fluorescently labelled ADEVs were added into primary cultured mouse cortical neurons, we found a significantly increased neuronal uptake of IL-1β-ADEVs compared to CTL-ADEVs. We further confirmed it is likely due to the enrichment of surface proteins in IL-1β-ADEVs, as IL-1β-ADEVs uptake by neurons was partially suppressed by a specific integrin inhibitor. Additionally, treatment of neurons with IL-1β-ADEVs also reduced neurite outgrowth, branching and neuronal firing. These findings provide insight for the molecular mechanism of the ADEVs’ effects on neural uptake, neural differentiation and maturation, and its alteration in inflammatory conditions.
KW - Astrocytes
KW - IL-1β
KW - exosome
KW - extracellular matrix
KW - extracellular vesicles
KW - inflammatory diseases
KW - neurodegenerative diseases
KW - proteomics
UR - http://www.scopus.com/inward/record.url?scp=85077158556&partnerID=8YFLogxK
U2 - 10.1080/20013078.2019.1706801
DO - 10.1080/20013078.2019.1706801
M3 - Article
AN - SCOPUS:85077158556
SN - 2001-3078
VL - 9
JO - Journal of Extracellular Vesicles
JF - Journal of Extracellular Vesicles
IS - 1
M1 - 1706801
ER -