Temporally resolved kinase regulatory networks control endothelial barrier integrity

Abstract

SUMMARYBreakdown of the blood-brain barrier is triggered by a range of physiological and pathological stimuli and is detrimental for brain function. Yet, the underlying signaling networks regulating barrier integrity are incompletely understood. Here, we present a novel and generalizable tool, Temporally REsolved KInase Network Generation (TREKING), that combines machine learning and network reconstruction to build time-resolved, functional phosphosignaling networks. We investigated kinase-driven pathways that modulate barrier permeability in brain endothelial cells in the presence of inflammatory stimuli. Our results reveal that >100 kinases are functional during barrier insult and provide time-resolved molecular insights into the differential networks that drive barrier disruption and recovery. The resulting models suggest a multi-layered rewiring of thrombin-induced barrier disruption following TNFα priming, including the timing of common signaling pathways, condition-specific phosphosignaling networks, and a skewing at major signaling hubs towards barrier-weakening activities. TREKING provides a novel tool for dissecting temporal phosphosignaling networks in biological systems.