Impact of N-acetyl cysteine ethyl ester on a-synuclein pathology in the olfactory system

N-乙酰半胱氨酸乙酯对嗅觉系统中α-突触核蛋白病理学的影响

基本信息

批准号：
8957097
负责人：
Rehana Khan Leak
金额：
$ 38.29万
依托单位：
DUQUESNE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8957097
关键词：
Address Alzheimer&apos s Disease Animals Apoptosis Astrocytes Atrophic Attenuated Award Biological Assay Biological Availability Blast Injuries Blinded Brain Cell Fractionation Cell Nucleus Cell Volumes Cells Clinical Trials Cognition Controlled Clinical Trials Cysteine Data Disease Dose Double-Blind Method Drug effect disorder Esters Etiology Experimental Models Family Family member Functional disorder Future Glutathione HSF1 Heat shock proteins Heat-Shock Proteins 70 Heat-Shock Response Human In Vitro Injection of therapeutic agent Light Link Literature Lysosomes MG132 Measures Mediating Messenger RNA Mitochondria Modeling Modification Molecular Mus Neurologic Neurons Nuclear Translocation Olfactory Pathways Parkinson Disease Pathology Patients Phosphorylation Placebo Control Proteasome Inhibitor Proteins Psychiatry RNA Interference Research Safety Serine Soldier Staining method Stains Stress Subfamily lentivirinae Sulfhydryl Compounds Symptoms Testing Thioflavin S Time Tissues Toxic effect Traumatic Brain Injury Ubiquitin Western Blotting Work acetaminophen overdose activating transcription factor alpha synuclein base bench to bedside cognitive function dietary supplements double-blind placebo controlled trial drinking water experience glucose-regulated proteins improved in vivo inhibitor/antagonist insight member mitochondrial heat shock protein 70 monomer multicatalytic endopeptidase complex neuroblastoma cell neuron loss novel olfactory bulb protective effect protein misfolding public health relevance response small hairpin RNA success synucleinopathy transcription factor

项目摘要

DESCRIPTION (provided by applicant): N-acetyl-L-cysteine (NAC) has been used to treat acetaminophen overdose since the 1970s and we already know that it has an excellent safety profile from these many decades of experience. NAC has also been used to treat patients with neurological conditions. For example, NAC was found to facilitate aspects of cognition in Alzheimer's disease. Furthermore, NAC was found to help resolve symptoms of blast injury in soldiers. However, questions have arisen about the low bioavailability of NAC and the need for high doses to penetrate the brain. We will develop use of the cell-permeable derivative, NAC ethyl ester (NACET), in a novel α-synuclein fibril-based model of Parkinson's disease. We will also establish the mechanism of protective action at the molecular level. Our pilot data on NAC already suggest that NAC can protect neurons and astrocytes in a heat shock protein-dependent manner. That is, NAC raises Hsp70A1/2 and pan-inhibitors of Hsp70 abolish its protective effects. This novel mechanism of NAC action has not been described by others in the literature, but may help explain why NAC has been effective in multiple double blind, placebo-controlled clinical trials. Here we will test the hypothesis that NACET can protect olfactory bulb neurons against α-synucleinopathy in an Hsp70-dependent manner. We have chosen to examine the olfactory bulb in these studies, as bulb atrophy and cell loss and olfactory dysfunction develop early in the course of Parkinson's disease. Aim 1a) Treat primary olfactory bulb neurons with a-synuclein fibrils and various concentrations of NACET. Determine if NACET protects against cell loss, raises Hsp70 family members, and mitigates an increase in ubiquitinated proteins and in Lewy-like inclusions. Aim 1b) Use 3 independent Hsp70 inhibitors to test if NACET-mediated protection is abolished with loss of Hsp70 activity. Aim 1c) Test if NACET raises Hsp70 levels by activating the transcription factors Nrf2 and HSF1. If Nrf2 or HSF1 is translocated to the nucleus after NACET treatment, knock it down with RNA interference to see if NACET-mediated protection is abolished. Aim 2a) Infuse α-synuclein fibrils into the mouse olfactory bulb and administer NACET in drinking water. Count NeuN+ neurons and Lewy-like inclusions by stereology after 6 months. Aim 2b) Repeat the fibril injections and NACET treatments and collect mouse tissue for Western blotting. Measure levels of Hsp70 family members and ubiquitinated proteins. Test the hypothesis that NACET will reduce neuron loss and Lewy-like inclusions in vivo and that it will raise Hsp70A1/2 and lower ubiquitinated proteins in olfactory bulb tissue. NACET may be the first treatment to safely boost Hsp70 defenses and reduce protein-misfolding stress in humans, with the potential to revolutionize Parkinson's treatment. These studies exemplify "bedside-to-bench" research, where compounds that are known to work in humans are examined post hoc in experimental models to yield deeper insights into disease etiology and drug action. This approach has been successful in the field of psychiatry for many years and it is time to leverage it in the battle against Parkinson's disease.

描述（由申请人提供）：自 20 世纪 70 年代以来，N-乙酰基-L-半胱氨酸 (NAC) 一直被用于治疗对乙酰氨基酚过量，并且从数十年的经验中我们已经知道它具有出色的安全性。例如，NAC 被发现可以促进阿尔茨海默病的认知功能，此外，NAC 还可以帮助解决士兵的爆炸损伤症状。由于 NAC 的生物利用度低以及需要高剂量才能渗透到大脑，我们将开发细胞渗透性衍生物 NAC 乙酯 (NACET) 在基于 α-突触核蛋白原纤维的帕金森病模型中的应用。我们还将在分子水平上建立保护作用机制。我们的 NAC 试验数据已经表明，NAC 可以以热休克蛋白依赖性方式保护神经元和星形胶质细胞。 Hsp70A1/2 和 Hsp70 的泛抑制剂消除了 NAC 的这种新作用机制，其他文献尚未对此进行描述，但可能有助于解释为什么 NAC 在多项双盲、安慰剂对照临床试验中有效。在这里，我们将测试 NACET 可以以 Hsp70 依赖性方式保护嗅球神经元免受 α-突触核蛋白病的影响，我们选择在这些研究中检查嗅球，如球萎缩。目标 1a) 用 α-突触核蛋白原纤维和不同浓度的 NACET 治疗初级嗅球神经元，确定 NACET 是否可以防止细胞丢失、增加 Hsp70 家族成员并减轻嗅觉功能障碍。泛素化蛋白和 Lewy 样内含物的增加目标 1b) 使用 3 种独立的 Hsp70 抑制剂来测试 NACET 是否介导。目标 1c) 测试 NACET 是否通过激活转录因子 Nrf2 和 HSF1 提高 Hsp70 水平。目标 2a) 将 α-突触核蛋白原纤维注入小鼠嗅球并施用 NACET。 6 个月后通过体视学对饮用水中的 NeuN+ 神经元和 Lewy 样内含物进行计数。目标 2b) 重复原纤维注射和 NACET 治疗并收集小鼠组织进行蛋白质印迹分析。 NACET 将减少体内神经元损失和路易样内含物，并提高 Hsp70A1/2 并降低嗅球中的泛素化蛋白NACET 可能是第一个能够安全增强人类 Hsp70 防御并减少蛋白质错误折叠应激的治疗方法，有可能彻底改变帕金森氏症的治疗方法。在实验模型中对人类进行事后检查，以更深入地了解疾病病因学和药物作用。多年来，这种方法在精神病学领域取得了成功，现在是时候在对抗帕金森病的斗争中利用它了。疾病。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Enhancing and Extending Biological Performance and Resilience.

DOI：
10.1177/1559325818784501
发表时间：
2018-07
期刊：
Dose-response : a publication of International Hormesis Society
影响因子：
0
作者：
Leak RK;Calabrese EJ;Kozumbo WJ;Gidday JM;Johnson TE;Mitchell JR;Ozaki CK;Wetzker R;Bast A;Belz RG;Bøtker HE;Koch S;Mattson MP;Simon RP;Jirtle RL;Andersen ME
通讯作者：
Andersen ME

Mechanistic Research for the Student or Educator (Part II of II).