Department of Biology and Volen Center for Complex Systems, Brandeis University, 415 South St., Waltham, MA 02453, USA.
Department of Biology and Volen Center for Complex Systems, Brandeis University, 415 South St., Waltham, MA 02453, USA.
Epilepsy Res. 2023 Jul;193:107156. doi: 10.1016/j.eplepsyres.2023.107156. Epub 2023 Apr 27.
Previously we demonstrated that intra-hippocampal infusion of purified, Semaphorin 4D (Sema4D) extracellular domain (ECD) into the mouse hippocampus rapidly promotes formation of GABAergic synapses and decreases seizure susceptibility in mice. Given the relatively fast action of Sema4D treatment revealed by these studies, we sought to determine the time course of Sema4D treatment on hippocampal network activity using an acute hippocampal slice preparation. We performed long-term extracellular recordings from area CA1 encompassing a 2-hour application of Sema4D and found that hippocampal excitation is suppressed for hours following treatment. We also asked if Sema4D treatment could ameliorate seizures in an acute seizure model: the kainic acid (KA) mouse model. We demonstrate that Sema4D treatment delays and suppresses ictal activity, delays the transition to Status Epilepticus (SE), and lessens the severity of SE. Lastly, we sought to explore alternative methods of Sema4D delivery to hippocampus and thus created an Adeno Associated Virus expressing the ECD of Sema4D. Our data reveal that virally delivered, chronically overexpressed Sema4D-ECD promotes GABAergic synapse formation and suppresses ictal activity and progression to SE. These results provide proof of concept that viral delivery of Sema4D is an efficacious and promising delivery method to abate epileptiform activity and progression to SE.
先前,我们证明了将纯化的 Sema4D(Semaphorin 4D)细胞外结构域(ECD)注入小鼠海马体中可快速促进 GABA 能突触的形成,并降低小鼠的癫痫易感性。鉴于 Sema4D 治疗的作用相对较快,我们希望通过急性海马切片制备来确定 Sema4D 治疗对海马体网络活动的时间进程。我们对包含 Sema4D 应用 2 小时的 CA1 区进行了长期的细胞外记录,发现治疗后数小时内海马体兴奋被抑制。我们还询问了 Sema4D 治疗是否可以改善急性癫痫模型中的癫痫发作:即红藻氨酸(KA)小鼠模型。我们证明,Sema4D 治疗可延迟和抑制癫痫发作,延迟向癫痫持续状态(SE)的转变,并减轻 SE 的严重程度。最后,我们试图探索向海马体输送 Sema4D 的替代方法,因此创建了一种表达 Sema4D ECD 的腺相关病毒(Adeno Associated Virus)。我们的数据表明,病毒介导的慢性过表达 Sema4D-ECD 可促进 GABA 能突触的形成,并抑制癫痫发作活动和向 SE 的进展。这些结果提供了概念验证,即病毒传递 Sema4D 是一种有效且有前途的方法,可以减轻癫痫样活动和向 SE 的进展。