Defining the MAST4 interactome in brain development and epilepsy

项目总结 癫痫是导致儿童和成人发病和死亡的常见且代价高昂的原因。发展和 癫痫脑病(DeE)是一种严重的儿童癫痫，需要快速诊断才能开始 控制癫痫发作，防止或逆转神经功能丧失的治疗。然而，这些 癫痫发作通常对药物没有反应。大多数DeE患者预计都有已知的潜在 遗传病因学，但他们往往缺乏分子诊断，因为潜在的致病变异是在 尚未与神经功能障碍相关的基因。我们已经确定了新的变种 MAST4基因与发育迟缓、DeE、视力异常以及一些结构性脑疾病患者的关系 异常现象。MAST4是一个研究较少的丝氨酸苏氨酸激酶家族的成员(MAST1-4和 Mast-like)，这是IDG合格的蛋白质之一，作为一种治疗手段，具有很高的影响人类健康的潜力 癫痫的目标。在目标1中，我们将表征MAST4在神经发育和神经发育中的时空表达 用无标记质谱仪鉴定体内MAST4相互作用蛋白。在目标2中，我们将验证 MAST4相互作用蛋白及患者特异性突变对这些相互作用的影响 体外培养神经元。这些实验的结果将提供重要的功能数据和 工具，指导具有MAST4突变的DIE患者获得精确医学的途径。

PROJECT SUMMARY Epilepsy is a common and costly cause of morbidity and mortality in children and adults. Developmental and epileptic encephalopathies (DEEs) are severe childhood epilepsies that require rapid diagnosis to initiate treatment that will control seizures and prevent or reverse the loss of neurologic function. However, these seizures often do not respond to medications. Most DEE patients are expected to have a known underlying genetic etiology, but they often lack a molecular diagnosis because the potential pathogenic variants are in genes that have not yet been associated with neurological dysfunction. We have identified de novo variants in the MAST4 gene in patients with developmental delay, DEE, vision abnormalities, and in some, structural brain abnormalities. MAST4 is a member of an understudied family of serine threonine kinases (MAST1-4 and MAST-like) that is among the IDG-eligible proteins with high potential to impact human health as a therapeutic target for epilepsy. In Aim 1, we will characterize MAST4 spatiotemporal expression in neurodevelopment and identify MAST4 interacting proteins in vivo using label-free mass spectrometry. In Aim 2, we will validate MAST4 interacting proteins and the impact of patient-specific mutations on these interactions in primary cultured neurons in vitro. The results from these experiments will provide both important functional data and tools to instruct the path to precision medicine for DEE patients with MAST4 mutations.