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.

摘要 帕金森病（PD）是一种进行性神经退行性疾病，影响1-2%的65岁以上的人。的 PD的经典运动症状是由脑实质中多巴胺能神经元的选择性变性引起的， 黑质旁核（SNpc），导致其长投射轴突输入纹状体的损失。电流 治疗策略[例如，多巴胺替代，脑深部电刺激（DBS）]只能最大限度地减少 黑质纹状体退化的症状，不能直接替代丢失的通路。因此，我们提出了一个新的 再生医学解决方案，其中定制的微组织工程神经网络（TENN）是 移植以物理地替代从SNpc到纹状体的轴突连接。具体来说，微- TENN将被移植到PD的啮齿动物和猪模型中，以直接替代SNpc神经元，恢复 轴突输入纹状体，并改善运动缺陷。我们的首要假设是， 由多巴胺能神经元和长的突出轴突束组成的微TENN将存活， 整合，从而在物理上重建黑质纹状体通路，以恢复运动功能的模型， 黑质纹状体传入阻滞为了验证这一假设，我们提出了三个目标：（1）确定最佳的体外 利用分化的神经元和干细胞， （2）评估micro-TENN重建黑质纹状体通路，恢复 多巴胺能输入，并改善啮齿动物的运动症状;（3）将人类规模的微型TENN应用于 重建猪的黑质纹状体通路。活体多巴胺能微TENNs将用 由具有单向长投射轴突束的离散神经元群组成的结构。 Micro-TENN的健康、表型、结构和功能将在体外进行优化。为了实现临床翻译， 我们将使用人类干细胞衍生的多巴胺能神经元构建人类规模的微型TENN。预成型 将构建体立体定向显微注射到神经退行性PD大鼠和猪模型中以评估回路 重建和运动症状改善。黑质纹状体通路重建将使用 行为、成像、电生理和组织学结果。拟议的工作将建立 个性化微型TENN的未来临床潜力，以改善PD运动症状， 多巴胺能黑质纹状体通路我们的微组织工程策略解决了临床 通过提供直接替代黑质纹状体通路的方法进行治疗，从而恢复运动 PD神经变性后的功能。由于它们的长轴突束，微TENN可能能够 替换退化的回路以恢复对纹状体的多巴胺能输入。我们的定制流程生成 微TENN能够实现精确工程化的结构，其中神经元的数量和神经元的生成 多巴胺可以在植入之前已知，因此，减轻了历史上在胎儿中看到的不一致问题。 组织移植因此，微TENN可以提供一种变革性和可扩展的解决方案，以永久地 取代失去的神经解剖结构，减轻数百万受 警局