Transcriptional and neuroprotective effects of hexokinase-2 inhibitors administered afterstroke

The inflammatory response induced by stroke can exacerbate injury to peri-infarct tissue. Microglia and other immune cells that mediate this response require increased glycolytic flux during pro-inflammatory activation. These cells, unlike neurons and most other cell types, utilize hexokinase-2 (HK2) rather than hexokinase-1 for glycolysis, such that HK2 inhibitors may selectively target them to suppress post-ischemic inflammation. Here we compared the effects of the non-selective hexokinase inhibitor 2-deoxyglucose to the HK2-selective inhibitors lonidamine and 3-bromopyruvate on secondary injury after stroke. A spatial transcriptomic assessment was performed in parallel to compare effects of the inhibitors on microglial gene expression and microglia - neuron interactions and to screen for off-target effects. Each of the inhibitors suppressed pro-inflammatory gene upregulation in peri-infarct microglia and attenuated the upregulation of cell stress functional pathways in the neighboring neurons, but had minimal effect on neuronal gene expression in uninjured cortex. The HK2-selective inhibitors were more effective than 2-deoxyglucose in suppressing morphological microglial changes, neuronal oxidative stress, and neurite loss. 3-bromopyruvate administered after stroke produced long-term improvements in functional outcome. Selective HK2 inhibitors may thus provide a clinically applicable means to suppress microglial activation and thereby improve outcomes after stroke without endangering neuronal energy metabolism.