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.

干细胞置换疗法是长期管理的有前途的治疗方法 与创伤或神经变性的运动缺陷相关的方法。但是，在细胞中 脊髓再生的疗法很有希望，迄今为止的研究患有不良神经元 集成和/或可变功能结果。原因之一可能是区域表型不匹配。 大脑中的研究强调了人多能干细胞区域规范的重要性 （HPSC）衍生的神经祖细胞减轻帕金森氏症，亨廷顿和癫痫病的症状 型号。脊髓中的可比研究受到控制区域的能力有限的阻碍 HPSC衍生的脊柱种群的表型。该提议的基本假设是 HPSC衍生的神经元移植对离散脊髓区域的遗传规范显着 影响植入功效，并随后患者的功能恢复。 这项工作的重点是运动神经元（MN），这些神经元专门针对多个 神经退行性疾病，脊髓损伤后受损。在神经管发育期间， Colinear Hox表达导致沿脊柱R/C轴的神经元表型的空间模式 绳索。阿什顿实验室已经建立了协议，概括了HPSC中的HOX进展，产生了神经 干细胞具有离散的HOX轮廓。当与腹膜构成的形态剂结合时，该方案 可以衍生出可以用作的祖细胞MN（PMN）和MN的完整延迟谱。 用于移植的区域特异性人群。 AIM 1专注于这些的产生和表征 高宫颈，中颈，肱，胸腔，腰部和s骨的区域化MN培养物代表 解剖段。除了通过分子和功能测定的表征，单细胞RNA-seq 将进行以确定区域化MN种群内的柱状和电动机池身份 移植。目标2和3检验了区域化HPSC衍生的PMN差异整合的假设 进入主机电路，并在体内有选择地增强功能恢复。 AIM 2检查PMN是否 优先植入其区域匹配的脊髓段，然后选择性地将轴突投射到 在发育小鸡模型中协调肌肉。 AIM 3试图确定是否有区域化PMN 移植到成年大鼠后，有助于功能恢复 phrenic Mns。期望是在移植沉默时会减轻行为的增长。在一起，这些 目的建立临床相关的MN种群进行移植，提高对机械的理解 人类MN多样化并确定区域特异性在神经元整合到中心的作用 神经系统。这些发现可以指导未来的临床干预措施，并可以翻译成其他脊柱细胞， 包括中间神经元和神经胶质。