Small Molecules that Activate Cytoprotective Functions of Tyrosyl-tRNA Synthetase

激活酪氨酰-tRNA 合成酶细胞保护功能的小分子

• 批准号: 10221719
• 负责人: Sajish Mathew
• 金额: $ 22.35万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-10 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221719
• 关键词: Active Sites; Affect; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Amino Acids; Amino Acyl-tRNA Synthetases; Amyloid beta-42; Amyloid beta-Protein; Area; Attenuated; Behavioral Assay; Binding; Biological Assay; Brain; Career Choice; Cell Nucleus; Centers of Research Excellence; Crystallization; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Repair Gene; DNA Repair Pathway; Data; Development; Drug Targeting; Enzymes; Evolution; Extramural Activities; Funding; Histologic; Homeostasis; Hydrogen Peroxide; Immunoblotting; In Vitro; Isomerism; Lead; Metabolic; Modeling; Molecular Conformation; Mus; N-Methylaspartate; Names; Nature; Neurodegenerative Disorders; Neuronal Survival Pathway; Neurons; Nicotinamide adenine dinucleotide; Nuclear; Oxidative Stress; Pathway interactions; Penetration; Phenols; Poly(ADP-ribose) Polymerases; Protein Biosynthesis; Proto-Oncogene Proteins c-akt; Publications; Publishing; Resveratrol; Series; Signal Transduction; Signal Transduction Pathway; Stress; Structure; Testing; Therapeutic; Tyrosine; Tyrosine-tRNA Ligase; Work; base; density; design; excitotoxicity; in vitro Model; in vitro activity; in vivo; in vivo evaluation; mouse model; nervous system disorder; neuroprotection; neurotoxicity; novel; novel drug class; novel strategies; oxidative damage; small molecule; targeted treatment; trans-resveratrol

ABSTRACT Aminoacyl-tRNA synthetases (aaRSs) are enzymes that activate L-amino acids for protein synthesis. However, during evolution, aaRSs progressively accrued ‘moonlighting’ functions that are activated under conditions of diminished protein synthesis and that enable aaRSs to regulate metabolic homeostasis and modulate signal transduction pathways. In particular, Tyrosyl-tRNA synthetase (TyrRS) moves into the nucleus under stress conditions and facilitates DNA repair through poly-ADP-ribose polymerase 1 (PARP1) activation. PARP1, the central regulator of nicotinamide adenine dinucleotide (NAD+) signaling, senses and responds to DNA damage and activates DNA repair pathways. Unrepaired or erroneously repaired DNA double strand breaks (DSBs) in neurons are a major contributing factor in the development of a variety of neurological disorders including Alzheimer’s disease (AD). In particular, accumulation of amyloid β(Aβ), a hallmark of AD, exacerbates the accumulation of DSBs in neurons. Remarkably, PARP1 is downregulated in AD and the TyrRS-PARP1 pathway was shown to attenuate Aβ-induced neurotoxicity. Stimulation of this pathway could offer a new approach to the treatment of AD and other neurodegenerative diseases. We hypothesize that the L-tyrosine binding pocket of eukaryotic TyrRS can be targeted to design and develop small molecules named TyrRS- Targeting Compounds (TTCs), which will activate the moonlighting functions of TyrRS that stimulate DNA repair, and that such molecules will exert neuroprotective activities in AD-relevant models. In the first year of this project, we have designed and synthesized a series of TTCs on the basis of our work published in Nature, which demonstrated that the cis-isomer of resveratrol (RSV) binds to TyrRS and acts as an L-tyrosine antagonist, activating PARP1-dependent signaling cascades. Some of the newly synthesized TTCs have already shown neuroprotective activity in vitro. Mechanistic analysis indicated that the lead neuroprotective compound activates AKT and upregulates DNA repair proteins. In the remaining period, we will pursue the following Specific Aims. Under Aim 1, we will continue structure-based design and synthesis of TyrRS Targeting Compounds (TTCs) based on the cis-RSV-bound co-crystal structure of TyrRS and demonstrate that such molecules engage and affect their TyrRS target. Under Aim 2, we will determine the effect of TTCs on the survival of primary cortical neurons and TyrRS/PARP1-dependent DNA repair and signaling cascades. Under Aim 3, we will test the in vivo therapeutic potential of TTCs that show neuroprotection in vitro using 5XFAD mouse model of AD. The completion of these Aims will provide proof-of-principle data that would guide the eventual development of a new class of drugs that targets TyrRS to activate PARP1-dependent DNA repair in neurons, with potential applicability to AD and other neurodegenerative diseases that originate from accumulated DNA damage.

抽象的 氨基酰基-TRNA合成酶（AARSS）是激活L-氨基酸以进行蛋白质合成的酶。然而， 在进化过程中，AARS逐渐获得的“月光”功能在 蛋白质合成减少，使AARS能够调节代谢稳态并调节信号 转导途径。特别是，在应激下，酪酶-TRNA合成酶（Tyrrs）进入核 条件并通过聚ADP-核糖聚合酶1（PARP1）激活促进DNA修复。 Parp1， 烟酰胺腺苷二核苷酸（NAD+）信号传导，对DNA损伤的反应 并激活DNA修复途径。未修复或错误修复的DNA双链断裂（DSB） 神经元是多种神经系统疾病的发展的主要因素 阿尔茨海默氏病（AD）。特别是，淀粉样β（Aβ）的积累，AD的标志，加剧了 DSB在神经元中的积累。值得注意的是，PARP1在AD和Tyrrs-Parp1中被下调 显示途径可减弱Aβ诱导的神经毒性。刺激这一途径可能会提供新的 治疗AD和其他神经退行性疾病的方法。我们假设L-酪氨酸 真核Tyrrs的结合袋可以针对设计和发展名为Tyrrs-的小分子 靶向化合物（TTC），它将激活刺激DNA的Tyrr的月光功能 维修，并且这种分子将在AD相关模型中执行神经保护活动。在第一年 这个项目，我们根据我们在自然界发表的工作设计并合成了一系列TTC， 这表明白藜芦醇（RSV）的顺式异构体与Tyrrs结合并充当L-酪氨酸 拮抗剂，激活PARP1依赖性信号级联。一些新合成的TTC具有 在体外已经显示出神经保护活性。机械分析表明铅神经保护 化合物激活AKT并上调DNA修复蛋白。在剩下的时期，我们将追求 遵循特定目标。在AIM 1下，我们将继续基于结构的设计和Tyrrs的合成 基于CIS-RSV结合的Tyrrs结构的靶向化合​​物（TTC），并证明这一点 这样的分子参与并影响其Tyrrs目标。在AIM 2下，我们将确定TTC对 原代皮质神经元和Tyrr/PARP1依赖性DNA修复和信号级联的存活。 在AIM 3下，我们将测试TTC的体内治疗潜力 5XFAD鼠标AD的模型。这些目标的完成将提供原则证明数据，以指导 针对Tyrrs激活PARP1依赖性DNA修复的新类药物的事件开发 在神经元中，具有起源于AD的潜在适用性和其他神经退行性疾病 累积的DNA损伤。