Human Dopamine Grafts in Alpha-Synuclein Models of Parkinson Disease

帕金森病α-突触核蛋白模型中的人多巴胺移植物

• 批准号: 10736403
• 负责人: VIVIANE TABAR
• 金额: $ 68.98万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736403
• 关键词: Ablation; Animal Model; Animals; Anti-Inflammatory Agents; Astrocytes; Behavior; Behavioral; Bilateral; Brain; Brain region; CSF1R gene; Candidate Disease Gene; Cell Lineage; Cells; Clinic; Clinical Trials; Collaborations; Complex; Corpus striatum structure; DNA; Data; Deposition; Derivation procedure; Deterioration; Development; Disease; Disease Progression; Disease model; Dopamine Agonists; Environment; Exhibits; Experimental Designs; Gene Expression Alteration; Gene Modified; Generations; Genes; Graft Survival; Histologic; Human; Human Engineering; Immunologic Deficiency Syndromes; Inflammation; Injections; Lewy Bodies; Longevity; Longitudinal Studies; Maps; Methods; Microglia; Midbrain structure; Modeling; Motor; Mus; Mutation; Natural regeneration; Nature; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neuronal Dysfunction; Neurons; Parkinson Disease; Pathogenesis; Pathogenicity; Patients; Persons; Phase I Clinical Trials; Phenotype; Population; Proteins; Regenerative Medicine; Research; Resistance; Resistance development; Role; SNCA gene; Safety; Signal Transduction; Substantia nigra structure; Techniques; Technology; Testing; Therapeutic; Time; Toxic effect; alpha synuclein; base editing; brain cell; cellular engineering; clinical lot; combinatorial; dopamine graft; dopaminergic neuron; glial activation; graft function; human embryonic stem cell; immunoregulation; in vivo; inhibitor; mitochondrial dysfunction; monomer; motor symptom; mouse model; mouse synuclein alpha; mutant; neuroinflammation; neuron loss; neuronal survival; neuropathology; neuroprotection; neurotoxic; neurotoxicity; neurotransmission; novel; pars compacta; pre-formed fibril; predictive modeling; prion-like; regeneration potential; restoration; stem cells; synucleinopathy; tool; transcriptomics; transmission process

ABSTRACT Parkinson's disease (PD) is one of the most common neurodegenerative disorders. It is characterized by the progressive loss of dopamine neurons in the substantia nigra (SN) pars compacta, and their projections onto striatal neurons and the accumulation of α-Synuclein aggregates often into Lewy bodies. The pathogenesis of PD is not fully elucidated but there is vast evidence supporting complex loops of neuroinflammatory cascades, mitochondrial dysfunction, degenerating neurons, sustained microglial activation and other pathophysiological mechanisms that together amplify a relentless progression towards neuronal loss in the nigra and beyond. α- Synuclein (α-S) aggregates are implicated either directly in serving as a template that is transmitted transneuronally and seeding further aggregation, and/or in amplifying the neuroinflammatory loop, leading in both cases to neuronal dysfunction and death. To date, there are no therapeutic options that lead to the regeneration of lost neurons or to the restoration of circuitry. Our group has pioneered the derivation of functional dopamine neurons from human embryonic stem cells (hES) and we have just completed a Phase 1 clinical trial for the bilateral intrastriatal grafting of these cells. There is much excitement about the restorative potential of stem cell derived neurons in PD, but there remain multiple challenges. Here we propose to study the impact of microenvironmental alterations in the brain in the context of 2 different mouse models: the 3K mouse model which expresses a triple mutant form of α-synuclein based on the human E46K mutation, and exhibits histological hallmarks of PD as well as progressive motor and other behavioral abnormalities; the second model consists of the intrastriatal injection of preformed α-synuclein fibrils (PFF) which spread transneuronally through the brain to form pathogenic α-synuclein inclusions, leading to loss of DA neurons and behavioral deterioration. These models are predicated on two different hypotheses and will serve as great tools to study inflammation and its impact on behavior. In addition, we will graft the mice with the same hES cell derived dopamine neurons used in the clinical trial to analyze the impact of the microenvironment on the neurons' survival and phenotype, as well as on their ability to rescue behavior. We will capture grafted cells as well as host microglia and astrocytes at key timepoints during the in vivo lifespan of the grafts to establish dynamic maps of cell lineages, maturation, microglial and astrocytic phenotypes and potentially activation of neurotoxic signals. In the last aim, we will engineer the human ES cells to delete the SNCA gene encoding α-synuclein in an attempt at increasing the resistance of the grafts to neurotoxicity and potentially the transmission of pathogenic α- synuclein. Data obtained in this proposal will serve to further enhance our understanding of neuro-inflammation in different PD microenvironments and could result in enhanced strategies for cell grafting including the use of gene edited cells that are resistant to neuroinflammation.

抽象的 帕金森氏病（PD）是最常见的神经退行性疾病之一。它以 在黑质（SN）pars commacta及其项目中逐渐丧失多巴胺神经元 纹状体神经元和α-突触核蛋白聚集体的积累经常进入Lewy身体。 PD的发病机理 没有完全阐明，但是有大量证据支持神经炎症级联反应的复杂环， 线粒体功能障碍，退化神经元，持续的小胶质细胞激活和其他病理生理学 共同扩大了尼格拉及其他地区神经元丧失的友好进展的机制。 α- 突触核蛋白（α-S）聚集体直接与传输的模板有关 跨神经和播种进一步聚集和/或放大神经炎性环，导致 这两个病例都遇到了神经元功能障碍和死亡。迄今为止，没有任何治疗选择导致 失去神经元的再生或回路的恢复。我们的小组率先推导了 来自人类胚胎干细胞（HES）的功能性多巴胺神经元，我们刚刚完成了1阶段1 这些细胞双侧双边移植的临床试验。恢复有很多兴奋 干细胞在PD中衍生的神经元的潜力，但仍有多个挑战。在这里，我们建议研究 在2种不同的小鼠模型的背景下，大脑中微环境改变的影响：3K小鼠 基于人E46K突变的α-突触核蛋白的三重突变形式的模型，并展示 PD的组织学标志以及渐进式运动和其他行为异常；第二个模型 由纹状体内注射预制的α-突触核蛋白原纤维（PFF），该核蛋白原纤维（PFF）在跨神经元扩散 通过大脑形成致病性α-突触核蛋白夹杂物，导致DA神经元和行为的丧失 恶化。这些模型在两个不同的假设上进行了预测，并将作为研究的绝佳工具 炎症及其对行为的影响。此外，我们将用同一HES细胞嫁接小鼠 在临床试验中使用的多巴胺神经元来分析微环境对神经元的影响 生存和表型，以及他们拯救行为的能力。我们将捕获嫁接的细胞以及宿主 在移植物的体内寿命期间，在关键时间点处的小胶质细胞和星形胶质细胞建立动态图 细胞谱系，成熟，小胶质细胞和星形细胞表型以及神经毒性信号的可能激活。 在最后一个目标中，我们将设计人类ES细胞以删除编码α-突触核蛋白的SNCA基因 增加移植物对神经毒性的抵抗力，并可能导致致病性α-的传播 综合蛋白。该提案中获得的数据将有助于进一步增强我们对神经炎症的理解 在不同的PD微环境中，可能会导致细胞移植的策略，包括使用 基因编辑的细胞对神经炎症具有抗性。