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（由适用提供）：自1970年代以来，N-乙酰L-半胱氨酸（NAC）已被用来治疗对乙酰氨基酚的过量剂，我们已经知道，从这些数十年的经验中，它具有出色的安全性。 NAC也已用于治疗患有神经系统疾病的患者。例如，发现NAC促进了阿尔茨海默氏病认知方面的方面。此外，发现NAC有助于解决士兵爆炸损伤的症状。然而，关于NAC的生物利用度低以及高剂量渗透大脑的需求引起了问题。我们将在基于帕金森氏病的新型α-突触核蛋白原纤维模型中开发使用可渗透的衍生物NAC乙基酯（NACET）。我们还将建立在分子水平保护作用的机制。我们关于NAC的试点数据已经表明，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。也就是说，NAC提高了HSP70A1/2，HSP70的泛抑制剂废除了其受保护作用。文献中其他人尚未描述这种NAC作用的新型机制，但可能有助于解释为什么NAC在多个双盲，安慰剂对照的临床试验中有效。在这里，我们将检验以下假设：NACET可以以HSP70依赖性方式保护嗅球囊神经元免受α-突触核病的影响。在这些研究中，我们选择检查嗅球，因为在帕金森氏病过程中早期就出现了鳞茎萎缩，细胞损失和嗅觉功能障碍。 AIM 1A）用A-核蛋白原纤维和各种浓度的NACET治疗原发性嗅球神经元。确定NACET是否可以预防细胞损失，增加HSP70家族成员，并减轻泛素化蛋白和Lewy样夹杂物的增加。 AIM 1B）使用3个独立的HSP70抑制剂来测试NACET介导的保护是否因HSP70活性的损失而被取消。 AIM 1C）测试NACET是否通过激活转录因子NRF2和HSF1提高HSP70水平。如果在NACET处理后将NRF2或HSF1转移到细胞核上，请在RNA干扰下将其击倒，以查看NACET介导的保护是否被废除。 AIM 2A）将α-突触核蛋白原纤维注入小鼠嗅球中，并在饮用水中施用NACET。 6个月后，计数Neun+神经元和类似Lewy的夹杂物。 AIM 2B）重复原纤维注射和NACET处理，并收集小鼠组织以进行蛋白质印迹。测量HSP70家族成员和泛素化蛋白的水平。检验了NACET将在体内减少神经元丧失和类似Lewy的夹杂物的假设，并且它将升高HSP70A1/2并降低嗅觉泡沫组织中的泛素化蛋白。 NACET可能是安全增强HSP70防御能力并减少人类蛋白质不满意的第一种治疗方法，并有可能改变帕金森氏症的治疗方法。这些研究举例说明了“床头到基础”的研究，其中在实验模型中检查了已知在人类中起作用的化合物，以产生对疾病病因学和药物作用的更深入的见解。多年来，这种方法在精神病学领域取得了成功，现在是时候在与帕金森氏病的战斗中利用它了。

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).