TY - JOUR
T1 - Mechanical stress induces Ca2+-dependent signal transduction in erythroblasts and modulates erythropoiesis
AU - Aglialoro, Francesca
AU - Abay, Asena
AU - Yagci, Nurcan
AU - Rab, Minke A. E.
AU - Kaestner, Lars
AU - van Wijk, Richard
AU - von Lindern, Marieke
AU - van den Akker, Emile
N1 - Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/1/2
Y1 - 2021/1/2
N2 - Bioreactors are increasingly implemented for large scale cultures of various mammalian cells, which requires optimization of culture conditions. Such upscaling is also required to produce red blood cells (RBC) for transfusion and therapy purposes. However, the physiological suitability of RBC cultures to be transferred to stirred bioreactors is not well understood. PIEZO1 is the most abundantly expressed known mechanosensor on erythroid cells. It is a cation channel that translates mechanical forces directly into a physiological response. We investigated signaling cascades down-stream of PIEZO1 activated upon transitioning stationary cultures to orbital shaking associated with mechanical stress, and compared the results to direct activation of PIEZO1 by the chemical agonist Yoda1. Erythroblasts subjected to orbital shaking displayed decreased proliferation, comparable to incubation in the presence of a low dose of Yoda1. Epo (Erythropoietin)-dependent STAT5 phos-phorylation, and Calcineurin-dependent NFAT dephosphorylation was enhanced. Phosphoryla-tion of ERK was also induced by both orbital shaking and Yoda1 treatment. Activation of these pathways was inhibited by intracellular Ca2+ chelation (BAPTA-AM) in the orbital shaker. Our results suggest that PIEZO1 is functional and could be activated by the mechanical forces in a biore-actor setup, and results in the induction of Ca2+-dependent signaling cascades regulating various aspects of erythropoiesis. With this study, we showed that Yoda1 treatment and mechanical stress induced via orbital shaking results in comparable activation of some Ca2+-dependent pathways, ex-hibiting that there are direct physiological outcomes of mechanical stress on erythroblasts.
AB - Bioreactors are increasingly implemented for large scale cultures of various mammalian cells, which requires optimization of culture conditions. Such upscaling is also required to produce red blood cells (RBC) for transfusion and therapy purposes. However, the physiological suitability of RBC cultures to be transferred to stirred bioreactors is not well understood. PIEZO1 is the most abundantly expressed known mechanosensor on erythroid cells. It is a cation channel that translates mechanical forces directly into a physiological response. We investigated signaling cascades down-stream of PIEZO1 activated upon transitioning stationary cultures to orbital shaking associated with mechanical stress, and compared the results to direct activation of PIEZO1 by the chemical agonist Yoda1. Erythroblasts subjected to orbital shaking displayed decreased proliferation, comparable to incubation in the presence of a low dose of Yoda1. Epo (Erythropoietin)-dependent STAT5 phos-phorylation, and Calcineurin-dependent NFAT dephosphorylation was enhanced. Phosphoryla-tion of ERK was also induced by both orbital shaking and Yoda1 treatment. Activation of these pathways was inhibited by intracellular Ca2+ chelation (BAPTA-AM) in the orbital shaker. Our results suggest that PIEZO1 is functional and could be activated by the mechanical forces in a biore-actor setup, and results in the induction of Ca2+-dependent signaling cascades regulating various aspects of erythropoiesis. With this study, we showed that Yoda1 treatment and mechanical stress induced via orbital shaking results in comparable activation of some Ca2+-dependent pathways, ex-hibiting that there are direct physiological outcomes of mechanical stress on erythroblasts.
KW - Calcium signal transduction
KW - Mechanical stress
KW - PIEZO1
UR - http://www.scopus.com/inward/record.url?scp=85100164715&partnerID=8YFLogxK
U2 - https://doi.org/10.3390/ijms22020955
DO - https://doi.org/10.3390/ijms22020955
M3 - Article
C2 - 33478008
SN - 1661-6596
VL - 22
SP - 1
EP - 15
JO - International journal of molecular sciences
JF - International journal of molecular sciences
IS - 2
M1 - 955
ER -