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Short and long TNF‐alpha exposure recapitulates canonical astrogliosis events in human‐induced pluripotent stem cells‐derived astrocytes
Author(s) -
Trindade Pablo,
Loiola Erick Correia,
Gasparotto Juciano,
Ribeiro Camila Tiefensee,
Cardozo Pablo Leal,
Devalle Sylvie,
Salerno José Alexandre,
Ornelas Isis Moraes,
Ledur Pitia Flores,
Ribeiro Fabiola Mara,
Ventura Ana Lucia Marques,
Moreira José Claudio Fonseca,
Gelain Daniel Pens,
Porciúncula Lisiane Oliveira,
Rehen Stevens Kastrup
Publication year - 2020
Publication title -
glia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.954
H-Index - 164
eISSN - 1098-1136
pISSN - 0894-1491
DOI - 10.1002/glia.23786
Subject(s) - astrogliosis , biology , induced pluripotent stem cell , microbiology and biotechnology , astrocyte , gliosis , glial fibrillary acidic protein , tumor necrosis factor alpha , neuroscience , immunolabeling , immunology , central nervous system , embryonic stem cell , genetics , immunohistochemistry , gene
Astrogliosis comprises a variety of changes in astrocytes that occur in a context‐specific manner, triggered by temporally diverse signaling events that vary with the nature and severity of brain insults. However, most mechanisms underlying astrogliosis were described using animals, which fail to reproduce some aspects of human astroglial signaling. Here, we report an in vitro model to study astrogliosis using human‐induced pluripotent stem cells (iPSC)‐derived astrocytes which replicate temporally intertwined aspects of reactive astrocytes in vivo. We analyzed the time course of astrogliosis by measuring nuclear translocation of NF‐kB, production of cytokines, changes in morphology and function of iPSC‐derived astrocytes exposed to TNF‐α. We observed NF‐kB p65 subunit nuclear translocation and increased gene expression of IL‐1β, IL‐6, and TNF‐α in the first hours following TNF‐α stimulation. After 24 hr, conditioned media from iPSC‐derived astrocytes exposed to TNF‐α exhibited increased secretion of inflammation‐related cytokines. After 5 days, TNF‐α‐stimulated cells presented a typical phenotype of astrogliosis such as increased immunolabeling of Vimentin and GFAP and nuclei with elongated shape and shrinkage. Moreover, ~50% decrease in aspartate uptake was observed during the time course of astrogliosis with no evident cell damage, suggesting astroglial dysfunction. Together, our results indicate that human iPSC‐derived astrocytes reproduce canonical events associated with astrogliosis in a time dependent fashion. The approach described here may contribute to a better understanding of mechanisms governing human astrogliosis with potential applicability as a platform to uncover novel biomarkers and drug targets to prevent or mitigate astrogliosis associated with human brain disorders.