Induced Pluripotent Stem Cells from Neurosurgical removed tissue for Brain Repair

来自神经外科切除组织的诱导多能干细胞用于大脑修复

• 批准号: 8445546
• 负责人: RAJ Raghavendra RAO
• 金额: $ 22.13万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8445546
• 关键词: Adult; Animals; Autologous; Biological; Bone Marrow; Brain; Cell Survival; Cell Transplants; Cells; Characteristics; Clinic; Development; Ectopic Expression; Emerging Technologies; Engineering; Environment; Epilepsy; Ethical Issues; Excision; Female; Fibroblasts; Generations; Goals; Graft Rejection; Graft Survival; Health; Healthcare; Human; Human Engineering; Immunocompetent; In Vitro; Lead; Methods; Mission; Motor; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Nude Rats; Outcome; Patients; Pluripotent Stem Cells; Population; Positioning Attribute; Property; Protocols documentation; Rattus; Replacement Therapy; Research; Research Personnel; Resources; Risk; Role; Sex Characteristics; Somatic Cell; Source; Stromal Cells; Techniques; Testing; The Sun; Tissues; Transplantation; Trauma; Traumatic Brain Injury; adult stem cell; base; brain repair; cell type; cognitive function; effective therapy; embryonic stem cell; fetal; improved; in vivo; induced pluripotent stem cell; injured; innovation; male; monolayer; nerve stem cell; nervous system disorder; neuronal replacement; novel; patch clamp; pluripotency; progenitor; relating to nervous system; repaired; research and development; self-renewal; senescence; stem; stem cell technology; tissue/cell culture; transcription factor; tumor; tumorigenesis

DESCRIPTION (provided by applicant): The long-term goal of this project is to develop patient-specific autologous cell source from induced pluripotent stem cells (iPSC) technology as a therapy for traumatic brain injury and other neurological diseases. Many current neural transplantation approaches have encountered problems associated with the source of donor cells. The ethnic controversies, limited cell availability, poor survival, lack of functional integration of grafted cells, the risk of tumor formation and immunological rejection in vivo are prominent issues need to be overcome before neural transplantation as a therapy to be used in clinic. It is essential, therefore, to identify optimal cell sources that are more conducive for cel replacement therapies. The emerging technology of human iPSCs generation, followed by directed differentiation into uniform populations of neural progenitor cells (hNPs) and neurons, holds great promise as an approach to reverse engineer human cells obtained from TBI patients. In this project, we are focusing our efforts on reprogramming adult human neural cells isolated from neurosurgical resection tissues. The specific hypothesis of this proposal is that functional iPSC-derived neural progenitor cells can be generated from neurosurgical resection tissues and utilized as cell replacement therapy for TBI. The hypothesis is based on our observations and research developments that (a) iPSCs can be generated from patients with specific neurodegenerative disorders, (b) directed differentiation of pluripotent stem cells can lead to uniform populations of hNPs, and (c) adult human stem-like cells can be isolated from neurosurgical resection tissues and successfully cultured as neurospheres and monolayers. To test the hypothesis, in this proposal, we will first generate iPSCs from adult human neural cells isolated from neurosurgical resection tissues and evaluate their pluripotent characteristics, directed differentiation capabilities in vitro. We will then assess the transplantation potential o these iPSC-derived hNPs in vivo in the brain in both intact immunodeficient animals and injured immunocompetent animals. The two proposed specific aims are: (1) Generate and characterize iPSCs from adult human neural cells isolated from neurosurgical resection tissues. (2) Evaluate the survival, differentiation and functionality of iPSC-derived NPs in the injured environment and assess the role of gender differences on cell survival. Results from the proposed study will have significant impact in the development of patient-specific hiPSCs as autologous cell replacement strategies for TBI and other neurological disorders. PUBLIC HEALTH RELEVANCE: Traumatic brain injury is fundamentally untreatable and clinically devastating. Generating induced pluripotent stem cells will significantly advance our strategies towards developing patient-specific cells for cell replacement therapy for TBI. The results from this proposal will provide positive impact on the development of autologous cell replacement therapies for TBI and other neurological diseases.

描述（由申请人提供）：该项目的长期目标是利用诱导多能干细胞（iPSC）技术开发患者特异性自体细胞来源，作为创伤性脑损伤和其他神经系统疾病的治疗方法。目前许多神经移植方法都遇到了与供体细胞来源相关的问题。种族争议、细胞可用性有限、存活率低、移植细胞缺乏功能整合、肿瘤形成风险和体内免疫排斥是神经移植作为一种治疗方法应用于临床之前需要克服的突出问题。因此，有必要确定更有利于细胞替代疗法的最佳细胞来源。人类 iPSC 生成的新兴技术，然后定向分化为统一的神经祖细胞 (hNP) 和神经元群体，作为对从 TBI 患者获得的人类细胞进行逆向工程的方法，前景广阔。在这个项目中，我们的重点是对从神经外科切除组织中分离出的成人神经细胞进行重新编程。该提案的具体假设是，功能性 iPSC 衍生的神经祖细胞可以从神经外科切除组织中产生，并用作 TBI 的细胞替代疗法。该假设基于我们的观察和研究进展：(a) iPSC 可以从患有特定神经退行性疾病的患者中产生，(b) 多能干细胞的定向分化可以产生均匀的 hNP 群体，(c) 可以从神经外科切除组织中分离出成人干细胞样细胞，并成功培养为神经球和单层细胞。为了检验这一假设，在本提案中，我们将首先从神经外科切除组织中分离出的成人神经细胞生成 iPSC，并评估其多能特性、体外定向分化能力。然后，我们将评估这些 iPSC 衍生的 hNP 在完整免疫缺陷动物和受伤的免疫功能动物大脑中的体内移植潜力。提出的两个具体目标是：（1）从神经外科切除组织中分离出的成人神经细胞生成 iPSC 并对其进行表征。 (2) 评估iPSC衍生的NP在受伤环境中的存活、分化和功能，并评估性别差异对细胞存活的作用。拟议研究的结果将对患者特异性 hiPSC 的开发产生重大影响，作为 TBI 和其他神经系统疾病的自体细胞替代策略。 公共卫生相关性：创伤性脑损伤基本上是无法治疗的，并且在临床上具有毁灭性。生成诱导多能干细胞将显着推进我们开发患者特异性细胞用于 TBI 细胞替代疗法的策略。该提案的结果将对 TBI 和其他神经系统疾病的自体细胞替代疗法的开发产生积极影响。