Treatment With Tetrahydrobiopterin Improves White Matter Maturation in a Mouse Model for Prenatal Hypoxia in Congenital Heart Disease

Abstract

Background Reduced oxygen delivery in congenital heart disease causes delayed brain maturation and white matter abnormalities in utero. No treatment currently exists. Tetrahydrobiopterin ( BH 4) is a cofactor for neuronal nitric oxide synthase. BH 4 availability is reduced upon NOS activation, such as during hypoxic conditions, and leads to toxin production. We hypothesize that BH 4 levels are depleted in the hypoxic brain and that BH4 replacement therapy mitigates the toxic effects of hypoxia on white matter. Methods and Results Transgenic mice were used to visualize oligodendrocytes. Hypoxia was introduced during a period of white matter development equivalent to the human third trimester. BH 4 was administered during hypoxia. BH 4 levels were depleted in the hypoxic brain by direct quantification (n=7–12). The proliferation (n=3–6), apoptosis (n=3–6), and developmental stage (n=5–8) of oligodendrocytes were determined immunohistologically. Total oligodendrocytes increased after hypoxia, consistent with hypoxia‐induced proliferation seen previously; however, mature oligodendrocytes were less prevalent in hypoxia, and there was accumulation of immature oligodendrocytes. BH 4 treatment improved the mature oligodendrocyte number such that it did not differ from normoxia, and accumulation of immature oligodendrocytes was not observed. These results persisted beyond the initial period of hypoxia (n=3–4). Apoptosis increased with hypoxia but decreased with BH 4 treatment to normoxic levels. White matter myelin levels decreased following hypoxia by western blot. BH 4 treatment normalized myelination (n=6–10). Hypoxia worsened sensory‐motor coordination on balance beam tasks, and BH 4 therapy normalized performance (n=5–9). Conclusions Suboptimal BH 4 levels influence hypoxic white matter abnormalities. Repurposing BH 4 for use during fetal brain development may limit white matter dysmaturation in congenital heart disease.