Functions of parkin in Parkinson’s disease

Parkin 在帕金森病中的作用

• 批准号: 9894863
• 负责人: JIAN FENG
• 金额: $ 45.03万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9894863
• 关键词: Action Potentials; Address; Adherent Culture; Affect; Axon; Basal Ganglia; Brain; Calcium Channel; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Corpus striatum structure; Development; Disease; Disease model; Exhibits; Fiber; Frequencies; Functional disorder; GIRK2 subunit, G protein-coupled inwardly-rectifying potassium channel; Genetic; Glutamates; Human; Inherited; Isogenic transplantation; Label; Lesion; Link; Locomotor Recovery; Methods; Midbrain structure; Molecular Target; Monkeys; Mutation; Neurons; Organoids; Parkinson Disease; Patients; Physiology; Preparation; Property; Rattus; Regulation; Role; Series; Substantia nigra structure; Symptoms; Synapses; System; Testing; Transplantation; base; cell type; dopaminergic neuron; genome editing; human embryonic stem cell; improved; in vivo; induced pluripotent stem cell; locomotor deficit; motor symptom; novel strategies; parkin gene/protein; repaired; stem cell technology; transmission process

Summary Nigral dopaminergic (DA) neurons (i.e. A9 DA neurons) that are lost in Parkinson’s disease (PD) have autonomous pacemaking action potentials, massive axon arborization, and expression of GIRK2, but not calbindin. Despite the significant progress in the differentiation of human embryonic stem cells (hESCs) to midbrain DA neurons, it has been difficult to generate A9 type DA neurons, particularly from human induced pluripotent stem cells (iPSCs). We developed an improved floorplate-based method to differentiate patient-specific iPSCs to midbrain DA neurons that expressed appropriate markers for A9 type cells and exhibited calcium channel-dependent autonomous pacemaking activities independent of glutamatergic inputs. These iPSC-derived DA neurons extended elaborate neuronal fibers when grafted to 6- OHDA-lesioned rats and restored locomotor deficits. We have generated isogenic pairs of iPSCs by repairing parkin mutations in patient cells and by introducing parkin mutations to control cells. Using these isogenic iPSCs, we will study how parkin mutations mechanistically disrupt the precision of dopaminergic transmission in three different preparations: monolayer cultures, brain organoids, and graft in 6-OHDA-lesioned rat brains. The three novel approaches will enable us to approximate the situation in the brains of PD patients. The study will bridge the gap between mechanistic understanding of the cellular function of parkin and its role in PD pathophysiology that is directly linked to the motor symptoms. The results will stimulate the development of disease-modifying therapies of Parkinson's disease .

概括 帕金森病中丢失的黑质多巴胺能 (DA) 神经元（即 A9 DA 神经元） 疾病（PD）具有自主起搏动作电位、大量轴突树枝化和 尽管在分化方面取得了显着进展，但 GIRK2 表达，但钙结合蛋白不表达。 人类胚胎干细胞（hESC）到中脑 DA 神经元，一直难以生成 A9 型 DA 神经元，特别是来自人类诱导多能干细胞 (iPSC)。 开发了一种改进的基于底板的方法来区分患者特异性 iPSC 中脑 DA 神经元表达 A9 型细胞的适当标记并表现出 独立于谷氨酸能的钙通道依赖性自主起搏活动 这些 iPSC 衍生的 DA 神经元在移植到 6- 后延伸出复杂的神经纤维。 我们已经生成了 OHDA 损伤的大鼠并恢复了运动缺陷。 iPSCs 通过修复患者细胞中的 Parkin 突变并将 Parkin 突变引入 使用这些同基因 iPSC，我们将研究 Parkin 突变的机制。 破坏三种不同制剂中多巴胺能传输的精度：单层 培养物、脑类器官和 6-OHDA 损伤大鼠大脑移植物这三种新方法。 将使我们能够大致了解帕金森病患者的大脑情况，这项研究将弥合这一问题。 Parkin 细胞功能的机制理解与其在 PD 中的作用之间存在差距 与运动症状直接相关的病理生理学结果将刺激。 帕金森病的疾病修饰疗法的开发。