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：神经茎或祖细胞的移植一直在增加，作为促进组织再生并重建TBI病变腔的有希望的策略。然而，在移植后，对人神经茎/祖细胞（HNSC）的分化的控制不佳，病变部位移植细胞的存活率低和整合已严重限制了基于细胞的疗法的成功。这项研究的目的是开发一种纳米粒子介导的转录激活方法，以将体内HNSC的分化指导为功能性神经元和少突胶质细胞。这种新方法将提供更好的控制和更高效率的生存，分化，成熟和体内功能细胞的整合。我们将通过将转录活化的HNSC在定制的透明质酸水凝胶中移植到创伤性脑损伤（TBI）的大鼠模型中，并检查细胞的存活，分化和分化移植细胞的分化以及在流向时期的组织再生结构。 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移植前人类诱导的多能干细胞（HIPSC）衍生的NSC将分别促进其在功能神经元和少突胶质细胞的体内分化。我们将在这项探索性赠款中追求以下具体目标：（1）通过编码转录因子NGN2和OLIG2的质粒分别建立一个高效的纳米颗粒来转染HIPSC衍生的NSC，并分别研究NGN2和Olig2表达对神经元和寡头endenderrocyte的效果。 （2）证明了源自体内HIPSC的NGN2转染和Olig2转染的HNSC的定向分化，以及水凝胶递送在促进转染的细胞的存活，分化和成熟和增强Rat Rat TBI模型中的组织再生方面的优势。这项研究不仅将使我们对转录因子激活的理解在调节体内移植的HIPSC衍生的NSC的生存，分化和成熟度中，而且还为治疗广泛的创伤性损伤和神经退行性疾病提供了新的策略。这种新的纳米颗粒方法以及具有定义成分的原位形成水凝胶非常需要临床翻译。