Directed In Vivo Differentiation of Neural Stem Cells for Repair of Brain Lesion

神经干细胞体内定向分化修复脑损伤

• 批准号: 8824696
• 负责人: Hai-Quan Mao
• 金额: $ 20.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8824696
• 关键词: Adopted; Brain; Cell Differentiation process; Cell Survival; Cell Therapy; Cell Transplantation; Cell Transplants; Cells; Cephalic; Clinical; Commit; Coupled; Effectiveness; Engraftment; Environment; Genetic Transcription; Grant; Growth Factor; Human; Hyaluronic Acid; Hydrogels; Implant; In Situ; In Vitro; Injectable; Injury; Lesion; Mediating; Methods; Modeling; Natural regeneration; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Oligodendroglia; Outcome; Patients; Plasmids; Production; Rattus; Recovery of Function; Site; Staging; Stem cell transplant; Stem cells; Testing; Tissues; Transcriptional Activation; Transcriptional Regulation; Transfection; Translations; Transplantation; Transplantation Conditioning; Traumatic Brain Injury; Viral; Viral Vector; angiogenesis; base; brain repair; brain tissue; cell type; controlled cortical impact; improved; in vivo; induced pluripotent stem cell; innovation; myelination; nanoparticle; nerve stem cell; nervous system disorder; novel strategies; oligodendrocyte lineage; public health relevance; relating to nervous system; small molecule; stem; stem cell differentiation; success; tissue regeneration; tissue repair; transcription factor; transgene expression

DESCRIPTION: Transplantation of neural stem or progenitor cells has been increasing proposed as a promising strategy to promote tissue regeneration and reconstruct the lesion cavity of TBI. However, poor control over the differentiation of human neural stem/progenitor cells (hNSCs) following transplantation, low survival and integration of the transplanted cells at the lesion site has severely limited the success of cell-based therapies. The objective of this study is to develop a nanoparticle-mediated transcription activation approach to direct the differentiation of hNSCs in vivo into functional neurons and oligodendrocytes. This new method will afford better control and higher efficiency of the survival, differentiation, maturation, and integration of the functional cells in vivo. We will demonstrate the advantages of this approach by transplanting transcriptionally activated hNSCs in a tailored hyaluronic acid hydrogel into a rat model of traumatic brain injury (TBI), and by examining cell survival, differentiation and integration of the transplanted cells, and tissue regeneration outcomes at the lesion site. Based on recent findings that over-expression of key transcriptional factors by viral transfection induces rapid and efficient production of functional neurons, and biodegradable nanoparticle transfection method capable of mediating efficient transgene expression in stem and progenitor cells, we plan to test the hypothesis that transient expression of stage-specific transcriptional factor neurogenin 2 (Ngn2) and oligodendrocyte transcription factor Olig2 in human induced pluripotent stem cell (hiPSC)-derived NSCs prior to transplantation will promote their in vivo differentiation towards functional neurons and oligodendrocytes, respectively. We will pursue the following specific aims in this exploratory grant: (1) to establish a highly effective nanoparticles for transfection of hiPSC-derived NSCs by plasmids encoding transcription factor Ngn2 and Olig2, respectively, and to investigate the effect of Ngn2 and Olig2 expression on neuronal and oligodendrocyte differentiation, respectively; (2) to demonstrate the directed differentiation of Ngn2-transfected and Olig2-transfected hNSCs derived from hiPSCs in vivo, and the advantage of hydrogel delivery in promoting the survival, differentiation, and maturation of the transfected cells and in enhancing tissue regeneration in a rat TBI model. This study will not only advance our understanding of transcription factor activation in regulating the survival, differentiation, and maturation of transplanted hiPSC-derived NSCs in vivo, but also offer new strategies for brain tissue regeneration in treating a wide range of traumatic injuries and neurodegenerative diseases. This new nanoparticle method together with the in situ forming hydrogels with defined compositions is highly desirable for clinical translation.

描述：越来越多的人提出神经干细胞或祖细胞移植作为促进组织再生和重建 TBI 病变腔的有前途的策略。然而，移植后对人类神经干/祖细胞（hNSC）分化的控制不佳，移植细胞在病变部位的存活率和整合度较低，严重限制了细胞疗法的成功。本研究的目的是开发一种纳米颗粒介导的转录激活方法，以指导 hNSC 在体内分化为功能性神经元和少突胶质细胞。这种新方法将为体内功能细胞的存活、分化、成熟和整合提供更好的控制和更高的效率。我们将通过将特制的透明质酸水凝胶中的转录激活的 hNSC 移植到创伤性脑损伤 (TBI) 的大鼠模型中，并通过检查细胞存活、移植细胞的分化和整合以及病变部位的组织再生结果来证明这种方法的优势。基于最近的发现，即通过病毒转染过度表达关键转录因子可诱导功能性神经元的快速高效产生，以及可生物降解的纳米颗粒转染方法能够介导干细胞和祖细胞中高效的转基因表达，我们计划检验阶段特异性转录因子神经原素 2 (Ngn2) 和少突胶质细胞转录因子 Olig2 瞬时表达的假设。 移植前的人诱导多能干细胞（hiPSC）衍生的 NSC 将促进其在体内分别分化为功能性神经元和少突胶质细胞。我们将在这笔探索性资助中追求以下具体目标：（1）建立一种高效纳米颗粒，用于分别编码转录因子Ngn2和Olig2的质粒转染hiPSC衍生的NSC，并研究Ngn2和Olig2表达分别对神经元和少突胶质细胞分化的影响； (2)在大鼠TBI模型中证明Ngn2转染和Olig2转染的hiPSC衍生的hNSC在体内定向分化，以及水凝胶递送在促进转染细胞的存活、分化和成熟以及增强组织再生方面的优势。这项研究不仅将增进我们对转录因子激活在调节体内移植的 hiPSC 衍生 NSC 的存活、分化和成熟方面的理解，而且还为治疗各种创伤性损伤和神经退行性疾病的脑组织再生提供新策略。这种新的纳米颗粒方法以及具有确定成分的原位形成水凝胶对于临床转化来说是非常理想的。