Rational derivation of DA neuron subtypes from iPS cells for improved modelling of Parkinson's disease

从 iPS 细胞中合理推导 DA 神经元亚型以改进帕金森病模型

基本信息

批准号：
9886284
负责人：
Rajeshwar B Awatramani
金额：
$ 56.65万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9886284
关键词：
Agonist Anatomy Back Cells Coupled Data Derivation procedure Development Disease Disease model Dopamine Dorsal Embryo Foundations Future Gene Expression Generations Genetic Genetic Models Genetic Transcription Goals Heterogeneity Human Individual Knowledge Label Laboratories Light Literature Logic Messenger RNA Methods Midbrain structure Modeling Molecular Molecular Profiling Mus Mutation Nerve Degeneration Neurons Parkinson Disease Pathologic Pathway interactions Patients Phenotype Physiological Physiology Production Property Protocols documentation Regimen Role SOX6 gene Solid Substantia nigra structure Sum Testing Time Tyrosine 3-Monooxygenase Ventral Tegmental Area WNT Signaling Pathway base beta catenin cohesion dopaminergic neuron dosage experimental study genetic manipulation human stem cells improved in vivo induced pluripotent stem cell loss of function mRNA Differential Displays motor symptom mouse model neuron development pars compacta postnatal progenitor public health relevance stem stem cell technology transcription factor transcriptome transcriptomics

项目摘要

DESCRIPTION (provided by applicant): Dopamine (DA) deficiency, caused by DA neuron degeneration, underpins the devastating motor symptoms of Parkinson's disease (PD). DA neurons located in the ventral tier of the substantia nigra pars compacta (SNc), are particularly vulnerable, compared to those in the dorsal tier of the SNc or ventral tegmental area (VTA). Why these DA neurons display differential vulnerability remains enigmatic. Understanding the underlying mechanisms would shed light on degeneration as well as potential neuroprotective strategies to mitigate the disease. iPS-derived DA neurons are an important new method for modeling PD. Yet current protocols for generating DA neurons are not designed to generate specific DA subtypes, a critical requisite for modeling selective vulnerability. This gap exists because the molecular heterogeneity of midbrain DA neurons is not well understood. To elucidate the heterogeneity of DA neurons, we have recently used single cell molecular profiling, coupled with anatomical co-labeling studies, and revealed the existence of at least six distinct of DA neuron subtypes in mouse models. Here, we aim to use this knowledge to i. better understand DA neuron diversity in vivo ii. understand mechanisms that may influence the generation of DA neuron subtypes iii. derive and characterize two prominent DA neuronal subtypes, one located in the SNc and one in the VTA, from human iPS cells in a rational manner, and iv. use these DA neuron subtypes to examine selective vulnerability in the context of genetic PD mutations. In Aim1, we will examine how Wnt signaling may influence DA neuron subtype allocation. In Aim 2, having optimized the Wnt regimen, we will next use targeted gene manipulations to derive highly enriched cultures of two specific DA neuron subtypes, and then characterize those subtypes by physiological and transcriptomic approaches. Next, we will generate both DA neuron subtypes from iPS cells harboring a DJ-1 mutation and examine differential pathological effects on both, SNc as well as VTA DA neuron subtypes. In Aim 3, we will further characterize the phenotype of the two DA neuron subtypes in vivo. We will elucidate the projections, and complete transcriptomes of two murine DA neuron subtypes, taking advantage of genetically targeted mice. Information from this aim will further highlight the differences between these subtypes. Additionally, these results will feed back into Aims 1 and 2, to further optimize our DA neuron subtype derivation protocol. In sum, taking advantage of the combined expertise and extensive interactions of two labs, we propose a cohesive plan based on molecular logic, to derive distinct DA neuron subtypes from iPS cells and aim to improve modelling PD. These studies will open the future possibility of understanding the effects of a range of PD mutations on selective vulnerability.

描述（由申请人提供）：由 DA 神经元变性引起的多巴胺 (DA) 缺乏是位于黑质致密部 (SNc) 腹侧层的帕金森病 (PD) 破坏性运动症状的基础。与 SNc 背层或腹侧被盖区 (VTA) 的神经元相比，这些 DA 神经元表现出不同的脆弱性的原因仍然存在。了解潜在的机制将有助于揭示退化现象以及缓解这种疾病的潜在神经保护策略，这是建模 PD 的重要新方法，但目前用于生成 DA 神经元的方案并非旨在生成特定的 DA。亚型，这是选择性脆弱性建模的关键必要条件。这种差距的存在是因为中脑 DA 神经元的分子异质性尚不清楚，为了阐明 DA 神经元的异质性，我们最近使用了单细胞分子分析，结合解剖学联合标记研究，揭示了至少六种不同的小鼠模型中的 DA 神经元亚型。在此，我们的目标是更好地了解体内 DA 神经元的多样性 ii. 了解可能影响 DA 神经元亚型生成的机制 iii. iv. 以合理的方式从人类 iPS 细胞中获取 SNc 中的一个和 VTA 中的一个，并使用这些 DA 神经元亚型来检查遗传 PD 突变背景下的选择性脆弱性。目标 1，我们将研究 Wnt 信号传导如何影响 DA 神经元亚型分配。在目标 2 中，优化 Wnt 方案后，我们接下来将使用靶向基因操作来获得两种特定 DA 神经元亚型的高度富集培养物，然后通过以下方式表征这些亚型。接下来，我们将从携带 DJ-1 突变的 iPS 细胞中生成两种 DA 神经元亚型，并检查对 SNc 和 VTA DA 神经元的不同病理影响。在目标 3 中，我们将利用来自该目标的遗传靶向信息进一步阐明两种小鼠 DA 神经元亚型的预测和完整转录组。此外，这些结果将反馈到目标 1 和 2，以进一步优化我们的 DA 神经元亚型推导协议。结合两个实验室的专业知识和广泛的互动，我们提出了一个基于分子逻辑的连贯计划，从 iPS 细胞中衍生出不同的 DA 神经元亚型，并旨在改进 PD 建模。这些研究将为理解一系列 DA 神经元的影响开启未来的可能性。 PD 突变对选择性脆弱性的影响。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Conversion of Quinazoline Modulators from Inhibitors to Activators of β-Glucocerebrosidase.

喹唑啉调节剂从β-葡萄糖脑苷脂酶抑制剂到激活剂的转换。

DOI：
10.1021/acs.jmedchem.8b01294
发表时间：
2019-01-15
期刊：
Journal of medicinal chemistry
影响因子：
7.3
作者：
Jianbin Zheng;S. Jeon;Weilan Jiang;Lena F. Burbulla;Daniel Ysselstein;Kristine Oevel;D. Krainc;R. Silverman
通讯作者：
R. Silverman

Organoid and pluripotent stem cells in Parkinson's disease modeling: an expert view on their value to drug discovery.

帕金森病模型中的类器官和多能干细胞：专家对其药物发现价值的看法。

DOI：
发表时间：
2020
期刊：
影响因子：
0
作者：
Marotta, Nick;Kim, Soojin;Krainc, Dimitri
通讯作者：
Krainc, Dimitri

Iron overload is accompanied by mitochondrial and lysosomal dysfunction in WDR45 mutant cells.

WDR45 突变细胞中铁超载伴随着线粒体和溶酶体功能障碍。

DOI：
发表时间：
2018-10-01
期刊：
影响因子：
0
作者：
Seibler, Philip;Burbulla, Lena F;Dulovic, Marija;Zittel, Simone;Heine, Johanne;Schmidt, Thomas;Rudolph, Franziska;Westenberger, Ana;Rakovic, Aleksandar;Münchau, Alexander;Krainc, Dimitri;Klein, Christine
通讯作者：
Klein, Christine