Characterization of secretomes from brain microvascular endothelial cells after ischemia

ABSTRACT: The human immortalized brain endothelial cell line hCMEC/D3 is considered an in vitro model of the blood-brain-barrier. We aimed to characterize changes in the secretome of hCMEC/D3 subjected to oxygen and glucose deprivation (OGD) by SILAC, in order to identify new proteins involved in ischemia-triggered blood-brain-barrier disruption and test their potential as blood biomarkers for ischemic stroke diagnosis. After SILAC analysis, 19 proteins were found differentially secreted between OGD and normoxia/normoglycemia conditions (Fold Change>|1.4| and peptide count≥2). Protein folding and nucleic acid binding were the main molecular functions and epithelial adherens junctions and aldosterone signaling appeared as the main canonical pathways represented by OGD-secreted proteins. ANXA1, CLUS, IGFBP2, PRDX3, TIMP2 and COL1A2 were replicated by western blotting in 9 independent cell cultures. Five replicated proteins were analyzed in human serum samples of 38 ischemic stroke patients compared to 18 stroke-mimicking conditions and 18 healthy controls by ELISA. IGFBP2 showed increased blood levels when strokes were compared with stroke-mimicking patients (p<0.1). In conclusion, we characterized changes in the secretome of hCMEC/D3 after an ischemic insult and highlighted some candidates to become biomarkers for ischemic stroke diagnosis related to blood-brain-barrier disruption.

INSTRUMENT(S): maXis

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

SUBMITTER: Víctor Llombart  

LAB HEAD: Joan Montaner

PROVIDER: PXD002292 | Pride | 2016-01-21

REPOSITORIES: Pride