Regionalized Human Motor Neuron Therapies

区域化人类运动神经元疗法

• 批准号: 9813519
• 负责人: Nisha Iyer
• 金额: $ 6.37万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-14 至 2021-09-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9813519
• 关键词: Ablation; Adult; Affect; Amyotrophic Lateral Sclerosis; Anatomy; Animal Model; Axon; Behavioral; Biocompatible Materials; Biological Assay; Brain; Breathing; Cell Line; Cell Survival; Cell Therapy; Cells; Cervical; Chemicals; Chest; Chick Embryo; Cholera Toxin; Clinical; Data; Derivation procedure; Development; Disease; Engraftment; Epilepsy; Event; Exhibits; Future; Gene Expression; Generations; Genetic; Grant; Homeobox Genes; Human; Huntington Disease; Hypoxia; Immunohistochemistry; Infection; Injections; Interneurons; Intervention; Location; Methods; Modeling; Molecular; Motor; Motor Neurons; Natural regeneration; Nerve Degeneration; Neural Tube Development; Neural tube; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neurons; Parkinson Disease; Patients; Pattern; Phenotype; Plethysmography; Population; Protocols documentation; Publishing; Rattus; Recovery; Recovery of Function; Rehabilitation therapy; Replacement Therapy; Reporting; Research; Rodent Model; Role; Sacral spinal cord structure; Source; Specificity; Spinal; Spinal Cord; Spinal Muscular Atrophy; Spinal cord injury; Stem cell transplant; Stem cells; Symptoms; Testing; Transgenic Organisms; Transplantation; Trauma; Work; base; cellular engineering; clinically relevant; clinically translatable; combinatorial; designer receptors exclusively activated by designer drugs; effective therapy; expectation; functional disability; functional outcomes; human pluripotent stem cell; improved; in vivo; locomotor deficit; morphogens; nerve stem cell; neuron loss; neuronal patterning; post-transplant; preference; progenitor; programs; rehabilitation strategy; relating to nervous system; respiratory; single-cell RNA sequencing; spinal cord regeneration

Stem cell replacement therapies are a promising, curative alternative to the long-term management approaches associated with locomotor deficits from trauma or neurodegeneration. However, while cell therapies for spinal cord regeneration are promising, studies to date have suffered from poor neuronal integration and/or variable functional outcomes. One reason for this may be regional phenotype mismatch. Studies in the brain have highlighted the importance of regional specification of human pluripotent stem cell (hPSC)-derived neural progenitors to alleviate Parkinson's, Huntington's, and Epilepsy" symptoms in rodent models. Comparable studies in the spinal cord have been hindered by a limited capacity to control the regional phenotype of hPSC-derived spinal populations. The fundamental hypothesis of this proposal is that genetic specification of hPSC-derived neuronal transplants to discrete spinal cord regions significantly affects engraftment efficacy and subsequently patients' functional recovery. This work focuses on motor neurons (MNs), which are specifically targeted in a number of neurodegenerative diseases and are damaged following spinal cord injury. During neural tube development, colinear HOX expression results in spatial patterning of neuronal phenotypes along the R/C axis of the spinal cord. The Ashton lab has established protocols recapitulate this Hox progression in hPSCs, generating neural stem cells with discrete Hox profiles. When combined with morphogens for ventral patterning, this protocol enables the derivation of a full rostrocaudal spectrum of progenitor MNs (pMNs) and MNs that can serve as region-specific populations for transplantation. Aim 1 focuses on the generation and characterization of these regionalized MN cultures representative of high cervical, mid cervical, brachial, thoracic, lumbar, and sacral anatomical segments. In addition to characterization by molecular and functional assays, single cell RNA-seq will be performed to determine columnar and motor pool identities within regionalized MN populations prior to transplantation. Aims 2 and 3 test the hypothesis that regionalized hPSC-derived pMNs differentially integrate into host circuits and selectively enhance functional recovery in vivo. Aim 2 examines whether pMNs preferentially engraft into their region-matched spinal cord segment and selectively project axons onto coordinate musculature in a developmental chick model. Aim 3 seeks to determine whether regionalized pMNs contribute to functional recovery following transplantation into an adult rat that has been selectively ablated of phrenic MNs. The expectation is that behavioral gains are mitigated upon transplant silencing. Together, these aims establish clinically relevant MN populations for transplantation, advance a mechanistic understanding of human MN diversification and establish the role of regional specificity on neuronal integration into the central nervous system. The findings can guide future clinical interventions and are translatable to other spinal cells, including interneurons and glia.

干细胞替代疗法是长期治疗的一种有前途、有疗效的替代疗法 与创伤或神经退行性疾病导致的运动缺陷相关的方法。然而，虽然细胞 脊髓再生疗法是有希望的，迄今为止的研究都受到神经元不良的影响 整合和/或可变的功能结果。造成这种情况的原因之一可能是区域表型不匹配。 大脑研究强调了人类多能干细胞区域规范的重要性 (hPSC) 衍生的神经祖细胞可缓解啮齿类动物的帕金森病、亨廷顿病和癫痫症状 模型。脊髓的可比研究因控制区域性神经元的能力有限而受到阻碍。 hPSC 衍生的脊髓群体的表型。该提案的基本假设是 hPSC 衍生的神经元移植到离散脊髓区域的遗传特异性显着 影响移植效果以及随后患者的功能恢复。 这项工作的重点是运动神经元（MN），它们专门针对许多 神经退行性疾病，并在脊髓损伤后受损。在神经管发育过程中， 共线 HOX 表达导致神经元表型沿脊柱 R/C 轴的空间模式 绳索。 Ashton 实验室已经建立了协议来概括 hPSC 中的 Hox 进展，从而产生神经 具有离散 Hox 谱的干细胞。当与腹侧图案形成的形态发生素结合时，该方案 能够推导祖 MN (pMN) 的完整 rostrocaudal 谱和可用作 特定区域的移植人群。目标 1 侧重于这些的生成和表征 代表高颈椎、中颈椎、肱椎、胸椎、腰椎和骶椎的区域化 MN 培养 解剖片段。除了通过分子和功能测定进行表征外，单细胞 RNA-seq 将在之前确定区域化 MN 人群中的柱状和运动池身份 移植。目标 2 和 3 检验区域化 hPSC 衍生的 pMN 差异整合的假设 进入宿主回路并选择性地增强体内功能恢复。目标 2 检查 pMN 是否 优先植入其区域匹配的脊髓段并选择性地将轴突投射到 协调发育雏鸡模型中的肌肉组织。目标 3 旨在确定区域化 pMN 是否 有助于移植到已选择性消融的成年大鼠体内后的功能恢复 膈 MN。预期行为增益会因移植沉默而减弱。在一起，这些 旨在建立临床相关的移植 MN 群体，增进对移植的机制的理解 人类 MN 多样化并建立区域特异性对神经元整合到中枢的作用 神经系统。这些发现可以指导未来的临床干预，并可转化为其他脊髓细胞， 包括中间神经元和神经胶质细胞。