The Rat Pre-Formed Alpha-Synuclein Fibril Model of Parkinson's Disease

帕金森病大鼠预制α-突触核蛋白原纤维模型

• 批准号: 9387180
• 负责人: Caryl E Sortwell
• 金额: $ 39.77万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9387180
• 关键词: Amygdaloid structure; Animal Model; Area; Behavior; Bilateral; Biological Models; Blood; Body Size; Clinical; Corpus striatum structure; Data; Denervation; Development; Diagnosis; Disease; Disease model; Dopamine; Drug Kinetics; Endopeptidase K; Exhibits; Female; Genetic; Human; Image; Impairment; Injectable; Injection of therapeutic agent; Intervention; Ipsilateral; Levodopa; Lewy Bodies; Light; Measurable; Memory impairment; Modeling; Monitor; Motor; Mus; Nerve Degeneration; Parkinson Disease; Pathologic; Pathology; Patients; Performance; Peripheral; Phase; Positron-Emission Tomography; Proteins; Rattus; Reporting; Resistance; Seeds; Substantia nigra structure; Synapses; Testing; Thioflavin S; Time; Tyrosine 3-Monooxygenase; Ubiquitin; Validation; alpha synuclein; brain size; clinical predictors; clinical translation; dopamine transporter; dopaminergic neuron; gender difference; in vivo; in vivo imaging; male; motor deficit; motor impairment; motor symptom; nerve supply; non-drug; novel; pars compacta; pharmacodynamic biomarker; pre-clinical; sample collection; synucleinopathy; treatment strategy; validation studies; vesicular monoamine transporter

Until recently, no animal model of Parkinson's disease (PD) has adequately incorporated both widespread alpha-synuclein (α-syn) pathology and protracted significant nigrostriatal degeneration. In 2012, Luk and colleagues described how intrastriatal injection of synthetic α-syn preformed fibrils (PFFs) into wildtype (WT) mice seeded endogenous accumulation of Lewy Body (LB)-like intracellular α-syn inclusions and ultimate nigrostriatal degeneration. In light of the fact that the rat model system offers distinct advantages over mice (fine motor behaviors, greater synaptic complexity, genetics and pharmacokinetics more similar to humans, larger brain and body size more amenable to neurosurgical interventions and sample collection), we recently characterized the results of unilateral injection of mouse α-syn PFFs into the striatum of rats. Similar to the mouse, we observed phosphorylated α-syn intraneuronal accumulations in several areas that innervate the striatum, most prominently the frontal and insular cortices, the amygdala, and the substantia nigra pars compacta (SNpc). α-Syn accumulations co-localized with ubiquitin, p62, and were thioflavin-S-positive and proteinase-k resistant. Although α-syn inclusions within the SNpc remained ipsilateral to striatal injection, we observed bilateral reductions in nigral dopamine neurons 6 months following α-syn PFF injection. Further, PFF injected rats exhibited reductions in striatal dopaminergic innervation as well as deficits in striatal dopamine and metabolites. This initial study demonstrates that α-syn PFFs are sufficient to seed the pathological conversion and propagation of endogenous α-syn to induce a progressive, neurodegenerative model of α-synucleinopathy in rats. In the present IGNITE application we seek to identify which α-syn species (human, mouse, rat) results in consistent loss of 60% nigral dopamine neurons over the course of 4 months after intrastriatal injection into rats (Aim 1). Once identified, we will conduct internal validation studies to systematically characterize the time course and magnitude of α-syn pathology, striatal dopamine and metabolite loss, levels of striatal dopaminergic innervation (tyrosine hydroxylase, vesicular monoamine transporter, and dopamine transporter) and deficits in motor behaviors (Aim 2). Lastly, we will conduct PD-relevant external validation studies including: 1) investigating whether PFF-induced motor deficits are reversible with levodopa and 2) conduct non-invasive longitudinal in vivo imaging of nigrostriatal DA synthesis, storage and turnover using positron emission tomography (PET) (Aim 3). We propose that optimization, internal validation and external validation studies in the progressive rat α-syn PFF PD model will allow for direct comparison to clinical metrics in PD patients and facilitate the development of novel disease modifying therapies.

直到最近，帕金森氏病（PD）的动物模型还充分纳入了宽度的α-突触核蛋白（α-syn）病理学和持久的明显的黑骨性变性。 2012年，Luk及其同事描述了合成α-Syn的纹状体内注射如何预先形成的原纤维（PFFS）到野生型（WT）小鼠中，将Lewy身体（LB）类细胞内α-α-α-α-α-α-α-α-同伴夹杂性和最终的Nigrostriptriatiation NigrostriatiataL的内源性积聚。 In light of the fact that the rat model system offers distinct advantages over mice (fine motor behaviors, greater synaptic complexity, genetics and pharmacokinetics more similar to humans, larger brain and body size more amenable to neurosurgical interventions and sample collection), we recently characterized the results of unilateral injection of mouse α-syn PFFs into the striatum of rats.与小鼠相似，我们观察到几种磷酸化的α-Syn神经内神经元积累，尽管SNPC内的α-SYN夹杂物仍然是纹状体注射的同侧，但我们观察到α-syn PFF注射6个月后6个月的双胺神经元的双侧减少。此外，PFF注射大鼠暴露于纹状体多巴胺能神经以及纹状体多巴胺和代谢物的碎片环境中。这项最初的研究表明，α-Syn PFF足以使内源性α-Syn的病理转化和繁殖以诱导大鼠α-Syn的进行性神经退行性模型。在目前的IGNITE应用中，我们试图确定哪些α-Syn物种（人，小鼠，大鼠）在注射到大鼠后4个月的时间内导致60％nigral多巴胺神经元持续损失（AIM 1）。一旦确定，我们将进行内部验证研究，以系统地表征α-SYN病理学，纹状体多巴胺和代谢产物的时间过程和大小，纹状体多巴胺能神经（酪氨酸羟化酶，囊心羟基单氨基单氨基单胺转运蛋白转运蛋白和多巴胺变速器）和多巴胺转移）和在Motory Prapisie in Morecii cardion cardies（Aim define criptiors）and（酪氨酸羟化酶）的水平。最后，我们将进行与PD相关的外部验证研究，包括：1）研究PFF诱导的运动定义是否可以用左旋多巴进行逆转和2）2）使用Polaron发射器（PET）（AIM 3）进行NIGROSTIRATAL DA合成，存储和传输的体内非侵入性纵向成像。我们建议，进行性大鼠α-Syn PFF PD模型中的优化，内部验证和外部验证研究将使PD患者的临床指标直接比较，并促进新的疾病修饰疗法的发展。