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Erythrocyte mitogen-activated protein kinases mediate hemolytic events under osmotic and oxidative stress and in hemolytic diseases
Journal article   Peer reviewed

Erythrocyte mitogen-activated protein kinases mediate hemolytic events under osmotic and oxidative stress and in hemolytic diseases

Kelsey Hazegh, Fang Fang, Kathleen Kelly, Derek Sinchar, Ling Wang, Benjamin E. Zuchelkowski, Alexander C. Ufelle, Orlando Esparza, Pavel Davizon-Castillo, Grier P. Page, …
Cellular signalling, Vol.99, pp.110450-110450
11/2022
DOI: 10.1016/j.cellsig.2022.110450
PMCID: PMC9530026
PMID: 36029940

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Abstract

p38 MAPKs are key regulators of cellular adaptation to various stress stimuli, however, their role in mediating erythrocyte cell death and hemolysis is largely unknown. We hypothesized that activation of erythrocyte p38 MAPK is a common event in the stimulation of hemolysis, and that inhibition of p38 MAPK pathways could mitigate hemolysis in hemoglobinopathies. We exposed human erythrocytes to diamide-induced oxidative stress or to hypoosmotic shock in the presence or absence of p38 MAPK inhibitors (SCIO469, SB203580, CMPD1) and used immunoblotting to determine MAPK activity and to identify possible downstream effectors of p38 MAPK. We also evaluated the impact of p38 MAPK inhibitors on stress-induced hemolysis or hypoxia-induced sickling in erythrocytes from mouse models of sickle cell disease. We found that human erythrocytes express conventional MAPKs (MKK3, p38 MAPK, MAPKAPK2) and identified differential MAPK activation pathways in each stress condition. Specifically, p38 MAPK inhibition in diamide-treated erythrocytes was associated with decreased phosphorylation of Src tyrosine kinases and Band 3 protein. Conversely, hypoosmotic shock induced MAPKAPK2 and RSK2 phosphorylation, which was inhibited by SCIO469 or CMPD1. Relevant to hemoglobinopathies, sickle cell disease was associated with increased erythrocyte MKK3, p38 MAPK, and MAPKAPK2 expression and phosphorylation as compared with erythrocytes from healthy individuals. Furthermore, p38 MAPK inhibition was associated with decreased hemolysis in response to diamide treatments or osmotic shock, and with decreased erythrocyte sickling under experimental hypoxia. These findings provided insights into MAPK-mediated signaling pathways that regulate erythrocyte function and hemolysis in response to extracellular stressors or human diseases. Visual abstract: Common and differential p38 MAPK signaling pathways that promote hemolysis under the tested stress conditions. Common to all evaluated stressors is the activation (phosphorylation, p) and membrane association of MKK3 and its downstream effector, p38 MAPK. A. Sickle cell disease and thalassemia are associated with increased expression of total and phosphorylated MKK3 and p38 MAPK in erythrocyte membranes compared with healthy controls. B. Diamide-induced oxidative stress promoted the activation of MKK3, p38 MAPK and erythrocyte Band 3 (anion exchanger-1). This pathway may involve interactions between p38 MAPK and Src proteins. C. Activation of p38 MAPK cascade under hypo-osmotic involved its downstream effector MAPAPK2 (MK2) and RSK2. D. Unlike hypo-osmotic stress, no changes were observed in MK2 and RSK2 membrane expression or phosphorylation under hyper-osmotic stress, however, p38 MAPK may promote Band 3 phosphorylation similar to the pathway depicted in B. graphics by Biorender.com. [Display omitted] •Human erythrocytes express conventional MAPKs including MKK3, p38 MAPK, and MK2.•Activation of MAPK cascades promotes hemolysis under osmotic and oxidative stress.•p38 MAPK expression is upregulated in hemolytic diseases.•p38 interacts with Src proteins to activate Band 3 under oxidative stress.•p38 MAPK inhibitors have antihemolytic and anti-sickling properties.
 Erythrocyte Hemolysis Oxidative stress p38 MAPK Sickle cell disease

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