Inhibiting pathological TDP-43 phosphorylation as a therapeutic strategy for ALS

TAR DNA结合蛋白43 kDa（TDP-43）是肌萎缩性侧索硬化症（AMO）患者的主要聚集性疾病蛋白。 侧索硬化症（ALS）。超过90%的ALS病例表现出含有洗涤剂的病理性病变 磷酸化、截短和泛素化TDP-43蛋白的不溶性沉积物。TDP-43 在S409/410磷酸化（pS409/410）是最一致的，稳健的，和特异性的神经病理学。 ALS的特征表明磷酸化TDP-43（pTDP）介导的神经毒性级联。 此外，pTDP已显示影响TDP-43在培养的细胞中的聚集和在细胞中的聚集。 人类ALS病例我们的前期工作证明了pS409/410 TDP-43介导运动神经元的运动 导致家族性ALS的TDP-43突变的毒性。存在调节TDP-43磷酸化的激酶 ALS治疗干预的一个有吸引力的目标。我们发现了一个保守的TDP-43- 具有翻译潜能的活性激酶，称为tau微管蛋白激酶1（TTBK 1）。脑识别 渗透TTBK 1抑制剂可能最终提供一个可行的药物开发策略。我们 假设TDP-43磷酸化增加驱动ALS中的运动神经元变性， 通过抑制TTBK 1来阻断pTDP积累将防止TDP-43介导的 ALS中的神经变性提出了三个综合的具体目标：1）确定TTBK 1 选择性激酶抑制剂。2)TTBK 1选择性抑制剂的优化和选择性抑制剂的验证 在pTDP积累的细胞模型中的化合物。3)验证TTBK 1在形成中的作用 使用TTBK 1敲除小鼠和现有的TDP-43蛋白质病转基因模型的pTDP。的 本文提出的研究将为TTBK 1选择性抑制剂的开发奠定基础 ALS的治疗方法

TAR DNA-binding protein 43 kDa (TDP-43) is the major aggregating disease protein in amyotrophic lateral sclerosis (ALS). Over 90% of ALS cases exhibit pathological lesions containing detergent insoluble deposits of phosphorylated, truncated, and ubiquitinated TDP-43 protein. TDP-43 phosphorylated at S409/410 (pS409/410) is the most consistent, robust, and specific neuropathological feature of ALS suggesting a phosphorylated TDP-43 (pTDP) mediated cascade of neurotoxicity. Furthermore pTDP has been shown to influence the aggregation of TDP-43 in cultured cells and in human ALS cases. Our previous work demonstrated pS409/410 TDP-43 mediates motor neuron toxicity of familial ALS-causing TDP-43 mutations. Kinases regulating TDP-43 phosphorylation present an attractive target for therapeutic intervention in ALS. We have identified a well-conserved TDP-43- active kinase with translational potential known as tau tubulin kinase 1 (TTBK1). Identification of brain penetrant TTBK1 inhibitors may ultimately provide a viable drug development strategy. We hypothesize that increased TDP-43 phosphorylation drives motor neuron degeneration in ALS and that blocking pTDP accumulation by inhibiting TTBK1 will protect against TDP-43 mediated neurodegeneration in ALS. Three integrated specific aims are proposed: 1) Identification of TTBK1 selective kinase inhibitors. 2) Optimization of TTBK1 selective inhibitors and validation of selective compounds in a cellular model of pTDP accumulation. 3) Validation of the role of TTBK1 in the formation of pTDP using TTBK1 knockout mice and an existing transgenic model of TDP-43 proteinopathy. The studies proposed here will set the stage for development of TTBK1 selective inhibitors as a candidate therapeutic approach for ALS.