Therapeutic evaluation of targeting Tau Tubulin kinase-1 in Alzheimer’s disease

靶向 Tau 微管蛋白激酶 1 治疗阿尔茨海默病的治疗评估

• 批准号: 10560906
• 负责人: Seiko Ikezu
• 金额: $ 54.78万
• 依托单位: MAYO CLINIC JACKSONVILLE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560906
• 关键词: Affect; Age Months; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease patient; Animal Model; Antisense Oligonucleotides; Arctic Regions; Autopsy; Axon; Biochemistry; Brain; CDK5 gene; Catalytic Domain; Cell Line; Cells; Central Nervous System; Chinese; Clinical Trials; Control Groups; Data; Dependovirus; Development; Episodic memory; FTD with parkinsonism; Functional Magnetic Resonance Imaging; Future; Goals; Hippocampus; Human; Immunohistochemistry; Impaired cognition; In Vitro; Induced pluripotent stem cell derived neurons; Injections; Knock-in; Knowledge; Late Onset Alzheimer Disease; Link; Medial; Mediator; Memory impairment; Mitosis; Modeling; Mus; Mutation; Nerve Degeneration; Neurons; Nucleic Acids; Odds Ratio; Organoids; Pathogenesis; Pathogenicity; Pathologic; Pathology; Patients; Persons; Phosphorylation; Phosphotransferases; Play; Protein Kinase; Publishing; Reagent; Reporting; Role; Semaphorin-3A; Senile dementia; Site; Specificity; Spinocerebellar Ataxias; Structure; Swedish mutation; System; Tauopathies; Temporal Lobe; Testing; Tg2576; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Thioflavin S; Transgenic Mice; Transgenic Organisms; axonal degeneration; behavior test; brain tissue; cilium biogenesis; cognitive function; cohort; comparison control; cross reactivity; drug development; effective therapy; efficacy evaluation; entorhinal cortex; experimental study; frontal lobe; hippocampal pyramidal neuron; in vivo; in vivo evaluation; induced pluripotent stem cell; kinase inhibitor; mild cognitive impairment; mouse model; neuron loss; neuropathology; novel; nucleic acid-based therapeutics; protein expression; response; safety assessment; stem cell model; tau Proteins; tau aggregation; tau dysfunction; tau expression; tau mutation; tau-1; tau-protein kinase; tau-tubulin kinase; therapeutic evaluation; therapeutic target; tool; transcriptome sequencing

Alzheimer’s disease (AD) is the most common form of senile dementia without effective treatments after countless number of clinical trials for over decades. This application proposes in vitro and in vivo evaluation of targeting human tau-tubulin kinase (TTBK1), a neuron specific tau kinase involved in the phosphorylation and aggregation of microtubule-associated protein tau in human and mice and genetically associated with late- onset AD in Spanish and Chinese cohorts. TTBK1 protein expression was significantly increased in the frontal cortex of postmortem AD brains compared to the control subject, suggesting that TTBK1 may play critical roles in AD pathogenesis. Our recent neuropathological examination of postmortem brain tissues revealed abundant expression of TTBK1 in the entorhinal cortex (EC) layer II and CA1 region of the hippocampal field where early tau pathology evolves in AD. Importantly, several studies reported that CA1 neuronal loss was correlated with cognitive decline in AD, and that CA1 connectivity to medial temporal cortex was associated with reduced episodic memory with mild cognitive impairment patients as determined by functional MRI. Interestingly, we found that TTBK1 significantly reduced axonal integrity of EC pyramidal neurons in AD mouse model, mimicking the early AD pathology. Thus, suppressing initial pathological tau accumulation in the EC and their spread to CA1 region by targeting TTBK1 could halt progression of AD at the prodromal stage. There are several attempts to develop TTBK1 kinase inhibitors, which were unsuccessful due to their cross-reactivity with TTBK2, a ubiquitous tau-tubulin kinase involved in mitosis and ciliogenesis. In this proposal, we will test the therapeutic specificity and efficiency of antisense oligonucleotides (ASOs) targeting TTBK1, which have little cross-reactivity to TTBK2. The central hypothesis of our proposal is that ASO-TTBK1 reduces accumulation and spread of pathological tau in early-stage AD brains. Our exciting preliminary data have shown that ASO- TTBK1 significantly reduced the level of phosphorylated tau at pSer396, pThr231 and pThr181 in the hippocampal region in PS19 tau mice. We will examine the therapeutic effect of ASO-TTBK1 on tau pathology and cognitive function in PS19 mice and our recently developed early tau propagation model (ECII-CA1 tau mice, Aim 1). We will further validate the effect of ASO-TTBK1 on tau pathology in human induced pluripotent stem cell-derived neurons and organoids isolated from late onset AD and FTDP-17 (P301L MAPT) cases in vitro (Aim 2) and on human TTBK1 transgenic and TTBK1:APP knock-in double transgenic mice in vivo (Aim 3). Successful completion of these experiments will validate TTBK1 as a therapeutic target of tauopathy and ASO as a translatable approach for targeting TTBK1 in AD brains.

阿尔茨海默氏病（AD）是老年痴呆症的最常见形式，在经历了严重的精神分裂症之后没有有效的治疗。 几十年来无数的临床试验。本申请提出了体外和体内评价 靶向人tau-微管蛋白激酶（TTBK 1），一种参与磷酸化的神经元特异性tau激酶， 微管相关蛋白tau在人类和小鼠中的聚集以及与晚期 西班牙和中国队列中的AD发作。TTBK 1蛋白表达在额叶皮层显著增加， 与对照受试者相比，死后AD大脑的皮质，这表明TTBK 1可能在其中起关键作用。 在AD发病机制中的作用我们最近对死后脑组织的神经病理学检查显示 TTBK 1在内嗅皮层（EC）第II层和海马区CA 1区的表达， tau病理学在AD中发展。重要的是，几项研究报道，CA 1神经元损失与 AD的认知能力下降，并且CA 1与内侧颞叶皮层的连接性与AD的认知能力下降有关。 情节记忆与轻度认知障碍患者的功能磁共振成像确定。有趣的是，我们 发现TTBK 1显著降低AD小鼠模型中EC锥体神经元的轴突完整性， 模仿早期AD病理学。因此，抑制EC中的初始病理性tau积累及其对细胞增殖的抑制作用是可能的。 通过靶向TTBK 1扩散到CA 1区，可以在前驱期阻止AD的进展。有 几次尝试开发TTBK 1激酶抑制剂，由于它们与 TTBK 2，一种参与有丝分裂和纤毛发生的普遍存在的tau-微管蛋白激酶。在本提案中，我们将测试 靶向TTBK 1的反义寡核苷酸（ASO）的治疗特异性和效率， TTBK 2的交叉反应性。我们建议的中心假设是ASO-TTBK 1减少了积累 以及病理性tau蛋白在早期AD大脑中的扩散。我们令人兴奋的初步数据表明，阿索- TTBK 1显著降低了pSer 396、pThr 231和pThr 181磷酸化tau蛋白的水平。 PS19 tau小鼠的海马区。我们将检查ASO-TTBK 1对tau病理学的治疗效果 以及我们最近开发的早期tau传播模型（ECII-CA 1 tau 小鼠，Aim 1）。我们将进一步验证ASO-TTBK 1在人诱导多能干细胞中对tau病理学的作用。 从迟发性AD和FTDP-17（P301 L MAPT）病例中分离的干细胞衍生的神经元和类器官， 体外（Aim 2）和体内对人TTBK 1转基因和TTBK 1：APP敲入双转基因小鼠（Aim （3）第三章。这些实验的成功完成将验证TTBK 1作为tau蛋白病的治疗靶点， 阿索作为在AD脑中靶向TTBK 1的可翻译方法。