Transplantable Micro-Tissue Engineered Neural Networks to Restore the Nigrostriatal Pathway in Parkinson's Disease

可移植微组织工程神经网络恢复帕金森病的黑质纹状体通路

• 批准号: 10403480
• 负责人: Daniel Kacy Cullen
• 金额: --
• 依托单位: PHILADELPHIA VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10403480
• 关键词: Address; Affect; Architecture; Axon; Behavioral; Brain; Characteristics; Clinical; Clinical Treatment; Corpus striatum structure; Custom; Deafferentation procedure; Deep Brain Stimulation; Disease model; Dopamine; Electrophysiology (science); Embryo; Engineering; Family suidae; Fetal Tissue Transplantation; Future; Generations; Health; Histologic; Human; Hydrogels; Image; Implant; In Vitro; Left; Maintenance; Medical center; Microinjections; Modeling; Motor; Nerve Degeneration; Nervous System Trauma; Nervous system structure; Neuroanatomy; Neurodegenerative Disorders; Neurons; Outcome; Parkinson Disease; Pathway interactions; Patients; Pennsylvania; Phenotype; Philadelphia; Population; Process; Rattus; Regenerative Medicine; Rodent; Rodent Model; Structure; Substantia nigra structure; Symptoms; Synapses; System; Techniques; Technology; Testing; Tissue Engineering; Tissues; Transplantation; Universities; Veterans; Work; base; brain circuitry; clinical translation; connectome; dopaminergic neuron; human adult stem cell; human stem cells; implantation; in vivo Model; motor deficit; motor symptom; multidisciplinary; neural network; neuronal replacement; neuronal survival; nigrostriatal dopaminergic pathway; nigrostriatal pathway; novel; optogenetics; pars compacta; porcine model; reconstruction; repaired; restoration; stem cell biology; stem cell derived tissues; stem cells; treatment strategy

ABSTRACT Parkinson's disease (PD) is a progressive neurodegenerative disease that affects 1-2% of people over 65. The classic motor symptoms of PD result from selective degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNpc), resulting in a loss of their long-projecting axonal inputs to the striatum. Current treatment strategies [e.g., dopamine replacement, deep brain stimulation (DBS)] can only minimize the symptoms of nigrostriatal degeneration, not directly replace the lost pathway. Therefore, we propose a novel regenerative medicine solution, whereby custom-built micro-tissue engineered neural networks (TENNs) are transplanted to physically replace the axonal connections from the SNpc to the striatum. Specifically, micro- TENNs will be transplanted in rodent and porcine models of PD to directly replace SNpc neurons, restore axonal inputs to the striatum, and ameliorate motor deficits. Our overarching hypothesis is that preformed micro-TENNs comprised of dopaminergic neurons and long-projecting axonal tracts will survive, synaptically integrate, and thereby physically reconstruct the nigrostriatal pathway to restore motor function in models of nigrostriatal deafferentation. To test this hypothesis, we propose three aims: (1) Determine optimal in vitro techniques to create dopaminergic micro-TENNs, using both differentiated neurons as well as stem-cell derived neurons; (2) Assess micro-TENN capabilities to reconstruct the nigrostriatal pathway, restore dopaminergic inputs, and ameliorate motor symptoms rodents; (3) Apply human-scale micro-TENNs to reconstruct the nigrostriatal pathway in swine. Living dopaminergic micro-TENNs will be constructed with an architecture consisting of a discrete population of neurons with unidirectional long-projecting axonal tracts. Micro-TENN health, phenotype, structure, and function will be optimized in vitro. To enable clinical translation, we will construct human-scale micro-TENNs using human stem cell derived dopaminergic neurons. Preformed constructs will be stereotactically microinjected into neurodegenerative PD rat and pig models to assess circuit reconstruction and motor symptom amelioration. Nigrostriatal pathway reconstruction will be assessed using behavioral, imaging, electrophysiological, and histological outcomes. The proposed work will establish the future clinical potential of personalized micro-TENNs to ameliorate PD motor symptoms by restoring the dopaminergic nigrostriatal pathway. Our micro-tissue engineering strategy addresses a crucial gap in clinical treatment by providing a means to directly replace the nigrostriatal pathway and, as a result, restore motor function following PD neurodegeneration. By virtue of their long axonal tracts, micro-TENNs may be capable of replacing degenerated circuitry to restore dopaminergic inputs to the striatum. Our custom process to generate micro-TENNs enables a precisely engineered structure where the number of neurons and generation of dopamine can be known prior to implantation, thus, alleviating issues of inconsistency historically seen in fetal tissue grafts. Therefore, micro-TENNs may provide a transformative and scalable solution to permanently replace lost neuroanatomy and alleviate the cause of motor symptoms for the millions of patient afflicted by PD.

抽象的 帕金森氏病（PD）是一种进行性神经退行性疾病，影响65岁以上的人中有1-2％。 PD的经典运动症状是由多巴胺能神经元选择性变性引起的 Nigra pars commacta（SNPC），导致其对纹状体的长期轴突输入损失。当前的 治疗策略[例如，多巴胺替代，深脑刺激（DBS）]只能最小化 黑质性变性的症状，不能直接替代丢失的途径。因此，我们提出了一本小说 再生医学解决方案，定制的微组织工程神经网络（TENN）是 移植以物理替代从SNPC到纹状体的轴突连接。具体而言，微型 - Tenn将在PD的啮齿动物和猪模型中移植，以直接替换SNPC神经元，恢复 轴突输入到纹状体，并改善运动缺陷。我们的总体假设是 由多巴胺能神经元和长注射的轴突段组成的微蛋 整合，从而物理地重建骨纹状体途径，以恢复模型中的运动功能 黑纹状体脱落。为了检验这一假设，我们提出了三个目的：（1）确定体外最佳 使用分化神经元和茎细胞创建多巴胺能微蛋白的技术 衍生的神经元； （2）评估重建骨纹状体途径的微元素能力，还原 多巴胺能输入和缓解运动症状啮齿动物； （3）将人尺度的微元素应用于 在猪中重建黑纹状体途径。活着的多巴胺能微型元素将由 由单向长期轴突区域的神经元组成的架构。 微蛋白健康，表型，结构和功能将在体外优化。为了启用临床翻译， 我们将使用人类干细胞衍生的多巴胺能神经元构建人尺度的微线。预先形成 构造将立体定位地微注射到神经退行性PD大鼠和猪模型中，以评估电路 重建和运动症状改善。将使用Nigrostriatal途径重建进行评估 行为，成像，电生理和组织学结果。拟议的工作将确定 个性化微元素的未来临床潜力通过恢复来改善PD运动症状 多巴胺能性黑质途径。我们的微组织工程策略解决了临床的关键差距 通过提供直接替换Nigrostriatal途径的手段来进行处理，并因此恢复电动机 PD神经变性后的功能。由于它们的长轴突缘，微镶嵌可能能够 取代退化的电路以将多巴胺能输入恢复到纹状体。我们生成的自定义过程 微型培养物具有精确的工程结构，神经元的数量和产生 多巴胺在植入之前可以知道，因此可以减轻胎儿历史上发现的不一致问题 组织移植。因此，微型围栏可以为永久性地提供变革性且可扩展的解决方案 替换失去的神经解剖学，减轻数百万患者的运动症状原因 PD。