Neural Progenitor Grafting for Restorative Stroke Therapy

用于恢复性中风治疗的神经祖细胞移植

• 批准号: 8214642
• 负责人: Jack M Parent
• 金额: $ 32.66万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8214642
• 关键词: Acute; Address; Adult; Allografting; Antigens; Area; Attention; Autologous; Bacterial Artificial Chromosomes; Behavior; Biological Assay; Brain; Brain Diseases; Cell Line; Cell Transplants; Cells; Corpus striatum structure; Cytogenetic Analysis; Development; Environment; Fibroblasts; Gene Expression Profiling; Genes; Goals; Green Fluorescent Proteins; Growth; Hippocampus (Brain); Human; Immunosuppression; In Vitro; Infarction; Injection of therapeutic agent; Ischemia; Knowledge; Light; Methods; Modeling; Natural regeneration; Neurons; Population; Property; Rattus; Recovery; Recovery of Function; Reporter; Risk; Safety; Skin; Somatic Cell; Source; Staging; Stem cell transplant; Stroke; Surface Antigens; Techniques; Teratoma; Testing; Transcript; Transplantation; Tubulin; Xenograft procedure; base; behavior influence; cell type; functional restoration; genetic manipulation; human embryonic stem cell; human embryonic stem cell line; improved; in vivo; induced pluripotent stem cell; injured; insight; migration; nerve stem cell; nervous system disorder; neurogenesis; neuronal replacement; novel; promoter; public health relevance; regenerative therapy; repaired; restorative treatment; self-renewal; stroke therapy; subventricular zone

DESCRIPTION (provided by applicant): The overall goals of this proposal are to determine the extent to which human embryonic stem cell (hESC)- or induced pluripotent stem cell (iPSC)-derived neural progenitor cell (NPC) grafts improve recovery after experimental stroke, and to begin addressing the critical safety and efficacy questions necessary for eventual human stroke therapy. hESCs offer many advantages as a source of NPCs for regenerative therapy, such as a readily available supply, vast differentiation potential and ease of genetic manipulation. iPSCs offer the added advantage of autologous grafting that obviates the need for immunosuppression. For either source, however, the ideal donor cell types and developmental states required to achieve CNS regeneration are poorly understood. Whether NPC grafting restores function after stroke and the mechanisms by which it might do so also are unknown. We have developed methods to enrich for specific NPC populations derived from hESCs, and have generated NPCs and multiple neuronal subtypes using iPSCs derived from human fibroblasts. Using these techniques, we propose to test the following hypotheses: 1) purified populations of multipotent NPCs (mpNPCs) and neuronal restricted precursors (NRPs) can be derived from hESCs or from human somatic cells via iPSCs. These populations will differ in their migration, differentiation and integration after transplantation into the intact or injured adult rat brain; and 2) grafting of mpNPCs or NRPs after experimental stroke will enhance functional recovery directly by neuronal replacement or by stimulating repair via endogenous NPCs. Specific Aims 1 and 2 are to purify and characterize specific NPC populations using promoter-based reporter or cell surface antigen-based selection. Aim 3 is to examine the behavior of these NPC populations after grafting into neurogenic and non-neurogenic regions of the adult rat brain, and Aim 4 is to examine the influence of hESC- and iPSC (human and rat)-derived NPC grafts on recovery after experimental stroke. Progress in these aims will advance our knowledge of how graft factors influence the capacity of hESC- or iPSC-derived NPCs to repair the injured brain and promote recovery after experimental stroke, and will provide insight into the untapped reparative potential and possible risks of these therapies. PUBLIC HEALTH RELEVANCE: Stroke is a common and potentially devastating neurological disorder with no proven regenerative therapies. This proposal aims to derive neural stem cells (NSCs) from human embryonic stem cells or reprogramming of human adult skin cells, and to use a stroke model to identify the optimal NSC grafts for brain reparative stroke therapy. Progress in this area offers advances toward novel cell-based restorative therapies for stroke and other brain insults.

描述（由申请人提供）：该提案的总体目标是确定人类胚胎干细胞（HESC）或诱导的多能干细胞（IPSC）衍生的神经祖细胞（NPC）移植物在实验性中进后改善恢复的恢复，并开始解决对最终人类疾病的重要安全性和效率的问题。 HESC作为NPC的再生疗法提供了许多优势，例如随时可用的供应，巨大的分化潜力和遗传操作的易用性。 IPSC提供了自体嫁接的额外优势，从而消除了对免疫抑制的需求。但是，对于任何一个来源，理想的供体细胞类型和实现中枢神经系统再生所需的发育状态尚不清楚。 NPC嫁接是否在中风后恢复功能以及可能这样做的机制也是未知的。我们开发了用于富含hESC的特定NPC种群的方法，并使用源自人成纤维细胞的IPSC生成了NPC和多个神经元亚型。使用这些技术，我们建议测试以下假设：1）可以通过IPSCS从hESC或人类体细胞中得出多能NPC（MPNPC）和神经元限制前体（NRP）的纯化群体。移植到完整或受伤的成年大鼠大脑后的迁移，分化和整合的迁移，分化和整合会有所不同。 2）实验中风后MPNPC或NRP的接枝将直接通过神经元替代或通过内源性NPC刺激修复。具体目的1和2是使用基于启动子的记者或基于细胞表面抗原的选择来纯化和表征特定的NPC种群。 AIM 3是检查成年大鼠脑的神经源性和非神经源性区域后这些NPC种群的行为，而AIM 4是检查hESC和IPSC（人类和大鼠）衍生的NPC移植物对实验性中风后恢复的影响。这些目标的进展将促进我们对移植因素如何影响hESC或IPSC衍生的NPC修复受伤大脑并促进实验性中风后恢复的能力的了解，并将洞悉这些疗法的未开发的修复潜力和可能的风险。 公共卫生相关性：中风是一种常见且潜在的毁灭性神经系统疾病，没有证实的再生疗法。该建议旨在从人类胚胎干细胞中得出神经干细胞（NSC）或重新编码人类成人皮肤细胞，并使用中风模型来鉴定最佳的NSC移植物来进行脑修复性中风治疗。该领域的进展为中风和其他大脑侮辱的新型基于细胞的修复疗法提供了进步。