WNKs regulate learning and memory in mice and alter neuronal glucose uptake and insulin signaling

Areas of the brain involved in spatial memory, such as the hippocampus, express high levels of insulin receptors and glucose transporter-4 (GLUT4) and are responsive to insulin. Insulin and neuronal glucose metabolism improve cognition in humans. Insulin-dependent GLUT4 trafficking has been extensively studied in muscle and adipose tissue, but little work has demonstrated either how it is controlled in insulin-responsive brain regions or its mechanistic connection to cognitive functions. In this study, we demonstrate that inhibition of the serine/threonine kinases WNKs (With-No-lysine (K)) using the pan-WNK inhibitor WNK463 improves learning and memory measured using fear conditioning test and the novel object recognition test in mice. WNK463 is a compound developed by Novartis and it is selective for WNKs over other enzymes based on a library of >400 protein kinases. Neuronal inhibition of WNK enhances in vivo hippocampal glucose uptake measured using uptake of tritiated 2-deoxyglucose. Inhibition of WNKs in primary neuronal cultures and differentiated neuroblastoma cell line SHSY5Y, enhances phosphorylated Protein kinase B (Akt) levels and thereby enhances the insulin signaling output. In addition, inhibition of WNKs increase insulin-dependent GLUT4 trafficking to the plasma membrane in mice primary neuronal cultures and hippocampal slices measured using surface biotinylation assay. Moreover, OSR1/SPAK kinases which are the best characterized downstream mediators of WNKs interact with critical molecular mediators involved in GLUT4 trafficking such as the AKT-substrate of 160-kDa (AS160) and sortilin. Therefore, we propose that neuronal WNK kinase activity has an important influence on learning and memory, in part, by modulation of neuronal insulin signaling. These studies were supported by NIH K99AG075161-01A1 to Ankita Jaykumar, NIH R01 HL147661 to Melanie Cobb, and 1R37NS114516-01A1 to Kim Huber. This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.