Kanzaki Sho, Shibata Shinsuke, Nakamura Masaya, Ozaki Masahiro, Okano Hideyuki
Department of Otolaryngology Head and Neck Surgery, Keio University School of Medicine, Tokyo, Japan.
Division of Hearing and Balance Research, National Institute of Sensory Organs, National Hospital Organization Tokyo Medical Center, Tokyo, Japan.
Methods Mol Biol. 2022;2524:327-332. doi: 10.1007/978-1-0716-2453-1_26.
Treatment for inner ear regeneration and protection needs local injection of steroid or new drug for inner ear regeneration into the round window membrane (RWM) in cochlea, but a systemic injection is not available due to its systemic side effects. However, pharmacokinetics of therapeutic agents or steroid locally injected into the inner ear is not well known. Hence, we introduce a new method for the real-time observation of drug delivery in transgenic animals in vivo. We exemplify mice which contain a firefly luciferase (FLuc) gene expression cassette regulated by the murine glial fibrillary acidic protein (GFAP) promoter. Luciferin delivered into the inner ear of those mice reacts with FLuc that is expressed in the GFAP-expressing cells in the cochlear nerve and spiral ganglion, and the resulting bioluminescence is detected by a camera. Using this system, we show the imaging of pharmacokinetic differences between local and systemic (intravenous and subcutaneous) injections of the inner ear.
内耳再生与保护的治疗需要将类固醇或用于内耳再生的新药局部注射到耳蜗的圆窗膜(RWM)中,但由于其全身副作用,无法进行全身注射。然而,局部注射到内耳的治疗剂或类固醇的药代动力学尚不清楚。因此,我们介绍一种在转基因动物体内实时观察药物递送的新方法。我们以含有由小鼠胶质纤维酸性蛋白(GFAP)启动子调控的萤火虫荧光素酶(FLuc)基因表达盒的小鼠为例。注入这些小鼠内耳的荧光素与在耳蜗神经和螺旋神经节中表达GFAP的细胞中表达的FLuc发生反应,产生的生物发光由相机检测。使用该系统,我们展示了内耳局部和全身(静脉内和皮下)注射药代动力学差异的成像。