Stem Cell-Derived Developmental Human Cortical Interneurons to Treat Intractable Epilepsy

干细胞衍生的发育性人类皮质中间神经元治疗难治性癫痫

• 批准号: 10355921
• 负责人: SANGMI CHUNG
• 金额: $ 56.58万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10355921
• 关键词: Address; Adult; Affect; Animal Model; Antiepileptic Agents; Behavioral; Brain; Cell Proliferation; Cell Survival; Cell Therapy; Clinical; Derivation procedure; Development; Devices; Disinhibition; Dose; Electroencephalography; Ensure; Environment; Epilepsy; Ethical Issues; Exhibits; Frequencies; GABA Agonists; Ganglia; Generations; Glutamates; Goals; Graft Survival; Growth; Hippocampus (Brain); Histologic; Homeostasis; Human; Image Analysis; Interneuron function; Interneurons; Intervention; Intractable Epilepsy; Investigational Therapies; Kainic Acid; Laboratories; Maintenance; Measures; Medial; Methods; Modeling; Monitor; Mus; Nervous system structure; Neurons; Patients; Pharmaceutical Preparations; Pilot Projects; Pluripotent Stem Cells; Population; Positioning Attribute; Rabies; Recurrence; Refractory; Reporting; Research; Risk; Safety; Seizures; Side; Specificity; Synapses; System; Temporal Lobe Epilepsy; Testing; Therapeutic; Three-Dimensional Image; Time; Translating; Translations; Transplantation; Work; brain circuitry; clinical application; clinical translation; comorbidity; density; effective therapy; efficacy evaluation; excitatory neuron; fetal; hippocampal sclerosis; human model; human pluripotent stem cell; in vivo imaging system; induced pluripotent stem cell; longitudinal analysis; migratory population; nerve supply; nervous system disorder; novel; novel therapeutics; postsynaptic; preclinical study; side effect; stem cells; tumor

Abstract Epilepsy is a severe neurological disease affecting more than 65 million people worldwide and is characterized by unpredictable abnormal electrical discharges resulting in recurrent seizures. About one third of patients with epilepsy suffer from intractable seizures that do not respond to antiepileptic drugs (AEDs). Neurosurgical interventions and neurostimulator devices are useful options for only a fraction of patients with drug-refractory seizures, underscoring the urgent need to develop new therapies. One strategy with considerable promise is to engraft new neurons to provide enhanced GABAergic inhibition in an activity-dependent manner. However, use of fetal neurons for cell therapy is associated with practical and ethical issues. Therefore, to overcome such hurdles, in our previous studies, we pioneered the transplantation of human pluripotent stem cells (hPSCs)- derived medial ganglionic eminence (MGE)-type human developmental cortical interneurons (cINs) into epileptic mouse brains and demonstrated their integration into dysfunctional circuitry, accompanied by the suppression of seizures and comorbid behavioral abnormalities. Furthermore, we have also determined the optimal stage of human cIN differentiation to ensure maximal integration into host circuitry as well as safety without risk of tumor formation, and developed a method to efficiently generate these safe and highly migratory populations of cINs from hPSCs in large quantities, bringing cell therapy for epilepsy one step closer to reality. However, there are still important issues to address prior to the clinical translation of this promising restorative therapy; 1) what is the synaptic connection specificity of human developmental cINs in adult epileptic circuitry? 2) what are safe and optimal densities of human cIN grafts for inhibition of epileptic host circuitry? 3) do human developmental cIN grafts maintain long-term efficacy and safety in epileptic brains? To tackle these issues, we will test our hypothesis that human iPSC-derived developmental cINs with optimal grafting densities preferentially innervate host excitatory neurons and ameliorate seizure activity with long-term efficacy and safety. We will transplant migratory human cINs into Nod Scid gamma (NSG) mice with intrahippocampal kainic acid-induced temporal lobe epilepsy (KA-TLE), a model of human hippocampal sclerosis, the most common cause of drug-resistant epilepsy, and analyze grafted cINs’ synaptic integration specificity and host inhibition in the epileptic brains. The long-term maintenance of anti-epileptic efficacy will be extensively analyzed by 24/7 video-EEG recordings 3 months, 6 months and 9 months after transplantation. We will analyze the grafts immunohistochemically to determine the extent of cell survival, maturation, integration, and most importantly, cell proliferation as a measure of graft safety without risk of uncontrolled growth. Completion of these studies is pivotal for translating this experimental therapy into a viable therapeutic strategy for intractable epilepsy.

摘要 癫痫是一种严重的神经系统疾病，影响全球超过6500万人，其特征是 无法预测的异常放电导致反复发作大约三分之一的患者 癫痫症患有对抗癫痫药物（AED）无反应的难治性癫痫发作。神经外科 介入和神经刺激器设备是有用的选择，只有一小部分患者的药物难治性 癫痫发作，强调迫切需要开发新的治疗方法。一个前景可观的战略是， 移植新的神经元以活性依赖性方式提供增强的GABA能抑制。但使用 胎儿神经元的细胞治疗与实际和伦理问题有关。为了克服这种 在我们以前的研究中，我们开创了人类多能干细胞（hPSC）的移植- 衍生的内侧神经节隆起（MGE）型人类发育皮层中间神经元（cIN）， 癫痫小鼠的大脑，并证明了他们的整合到功能失调的电路，伴随着 抑制癫痫发作和共病行为异常。此外，我们还确定了 人类cIN分化的最佳阶段，以确保最大程度地整合到宿主电路中以及安全性 没有肿瘤形成的风险，并开发了一种方法，有效地产生这些安全和高度迁移的 从hPSC中大量提取cIN群体，使癫痫的细胞治疗更接近现实。 然而，在这种有前途的修复体的临床转化之前，仍然有重要的问题需要解决。 治疗; 1）成人癫痫回路中人类发育cINs的突触连接特异性是什么？ 2)人cIN移植物抑制癫痫宿主回路的安全和最佳密度是多少？3)进行人体 发育cIN移植物在癫痫脑中保持长期有效性和安全性？为了解决这些问题，我们 将检验我们的假设，即具有最佳移植密度的人iPSC衍生的发育cIN 优先支配宿主兴奋性神经元，并长期有效地改善癫痫发作活动， 安全为代价的我们将迁移性人cIN移植到Nod Scid γ（NSG）小鼠中， 海人酸诱导的颞叶癫痫（KA-TLE）是人类海马硬化的模型， 耐药癫痫的常见病因，并分析移植cINs的突触整合特异性和宿主 抑制癫痫的大脑。抗癫痫疗效的长期维持将广泛 在移植后3个月、6个月和9个月通过24/7视频EEG记录进行分析。我们将 化学分析移植物以确定细胞存活、成熟、整合的程度， 最重要的是，细胞增殖作为移植物安全性的量度，而没有不受控制的生长的风险。完成 这些研究对于将这种实验性疗法转化为可行的治疗策略至关重要， 顽固性癫痫