Departments of Neurobiology (J.A.J., C.-Y.Y., K.A.H.-S., A.J.S., G.J.K., S.M., E.A.) and Pharmacology and Chemical Biology (D.T.M., M.J.P.) and Pittsburgh Institute for Neurodegenerative Diseases (J.A.J., D.T.M., C.-Y.Y., K.A.H.-S., A.J.S., G.J.K., S.M., M.J.P., E.A.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
Departments of Neurobiology (J.A.J., C.-Y.Y., K.A.H.-S., A.J.S., G.J.K., S.M., E.A.) and Pharmacology and Chemical Biology (D.T.M., M.J.P.) and Pittsburgh Institute for Neurodegenerative Diseases (J.A.J., D.T.M., C.-Y.Y., K.A.H.-S., A.J.S., G.J.K., S.M., M.J.P., E.A.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
J Pharmacol Exp Ther. 2018 Nov;367(2):348-355. doi: 10.1124/jpet.118.252338. Epub 2018 Sep 6.
We present the design of an innovative molecular neuroprotective strategy and provide proof-of-concept for its implementation, relying on the injury-mediated activation of an ectopic gene construct. As oxidative injury leads to the intracellular liberation of zinc, we hypothesize that tapping onto the zinc-activated metal regulatory element () transcription factor 1 system to drive expression of the Kv2.1-targeted hepatitis C protein NS5A (hepatitis C nonstructural protein 5A) will provide neuroprotection by preventing cell death-enabling cellular potassium loss in rat cortical neurons in vitro. Indeed, using biochemical and morphologic assays, we demonstrate rapid expression of -driven products in neurons. Further, we report that -driven NS5A expression, induced by a slowly evolving excitotoxic stimulus, functionally blocks injurious, enhanced Kv2.1 potassium whole-cell currents and improves neuronal viability. We suggest this form of "on-demand" neuroprotection could provide the basis for a tenable therapeutic strategy to prevent neuronal cell death in neurodegeneration.
我们提出了一种创新的分子神经保护策略的设计，并提供了其实验概念的证据，该策略依赖于损伤介导的异位基因构建的激活。由于氧化损伤导致细胞内锌的释放，我们假设利用锌激活的金属调节元件（）转录因子 1 系统来驱动靶向 Kv2.1 的肝炎 C 蛋白 NS5A（肝炎 C 非结构蛋白 5A）的表达，将通过防止细胞死亡来提供神经保护，从而使体外培养的大鼠皮质神经元中的细胞内钾丢失。事实上，我们使用生化和形态学测定方法证明了在神经元中快速表达 - 驱动产物。此外，我们报告说，由缓慢发展的兴奋性毒性刺激诱导的 - 驱动的 NS5A 表达，可有效地阻断有害的、增强的 Kv2.1 钾全细胞电流，并提高神经元活力。我们认为这种形式的“按需”神经保护可以为预防神经退行性变中的神经元细胞死亡提供可行的治疗策略的基础。
