Induced Pluripotent Stem Cells from Neurosurgical removed tissue for Brain Repair

描述(申请人提供)：该项目的长期目标是从诱导多能干细胞(IPSC)技术中开发患者特定的自体细胞来源，作为创伤性脑损伤和其他神经疾病的治疗方法。目前的许多神经移植方法都遇到了与供体细胞来源相关的问题。在将神经移植作为一种治疗方法应用于临床之前，种族争议、细胞可获得性有限、存活率低、移植细胞缺乏功能整合、体内肿瘤形成的风险和免疫排斥反应是需要克服的突出问题。因此，确定更有利于细胞替代疗法的最佳细胞来源是至关重要的。新出现的人类IPSCs生成技术，随后定向分化为统一的神经前体细胞(HNPs)和神经元群体，有望成为反向工程从脑外伤患者获得的人类细胞的方法。在这个项目中，我们集中精力对从神经外科手术切除组织中分离出来的成人神经细胞进行重新编程。这一建议的具体假设是，从神经外科手术切除的组织中可以产生功能性IPSC来源的神经前体细胞，并将其用于脑外伤的细胞替代治疗。这一假说是基于我们的观察和研究进展，即(A)IPSCs可以从特定的神经退行性疾病患者身上产生，(B)多能干细胞的定向分化可以导致hNPs群体的统一，以及(C)成人干细胞可以从神经外科手术切除组织中分离出来，并成功培养为神经球和单层细胞。为了验证这一假设，在这项提议中，我们将首先从神经外科手术切除组织中分离的成人神经细胞中产生IPSCs，并评估其体外定向分化的多能性特征。然后，我们将评估这些IPSC衍生的hNPs在完整的免疫缺陷动物和受损的免疫活性动物体内脑内的移植潜力。提出的两个具体目标是：(1)从神经外科手术切除组织中分离的成人神经细胞中培养并鉴定IPSCs。(2)评估IPSC来源的NPs在损伤环境中的存活、分化和功能，并评估性别差异对细胞存活的作用。这项拟议的研究结果将对发展患者特有的HiPSCs作为治疗脑外伤和其他神经疾病的自体细胞替代策略产生重大影响。 公共卫生相关性：创伤性脑损伤从根本上说是无法治疗的，在临床上是毁灭性的。产生诱导的多能干细胞将极大地推进我们的战略，为脑外伤的细胞替代治疗开发针对患者的细胞。这项建议的结果将对开发治疗脑损伤和其他神经系统疾病的自体细胞替代疗法产生积极影响。

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.