STEM/PROGENITOR CELL PROTECTION FOR PARKINSON'S DISEASE

描述（申请人提供）：有必要为神经退行性疾病中的高危神经元开发细胞和分子保护方法，再生医学提供了设计细胞，用于帕金森病（PD）等疾病的因子递送，我们知道某些生长因子如果递送得当，有能力拯救黑质纹状体细胞的离散群体，从而减缓或停止疾病的进展。该提案将使用一种新的细胞移植方法来提供胶质细胞系衍生的神经营养因子（GDNF）在可比的多巴胺神经元变性的体外和体内生物测定。三个具体的目标集中在哪些星形胶质细胞，从不同的来源，可以用来提供GDNF的风险多巴胺神经元在培养模型的PD以及广泛接受的啮齿动物模型（6-羟基多巴胺，6- OHDA病变的新纹状体）的疾病。具体目标1将通过关注源自小鼠大脑（例如成年大脑皮层和纹状体）的新生成的“未成熟”星形胶质细胞以及骨髓与小鼠胚胎干细胞来建立自体细胞修复的潜力。这些候选星形胶质细胞群将用eGFP-GDNF慢病毒构建体转导，以确定哪一个或哪些群体最适合于稳定的生长因子转导和生长因子的释放，用于随后在体内损伤保护模型中的测试（目的3）。具体目标2将着眼于成年人脑星形细胞和骨髓来源的细胞，以及它们稳定地用lenti-GDNF转导并释放生长因子的能力，就像目标1中的小鼠细胞那样。Specific Aim 3将测定释放GDNF的小鼠和人细胞在DA耗竭小鼠中的潜在保护作用。这些研究将结合我们实验室已经开发和测试的方法，包括细胞培养，电生理学，基因治疗，细胞移植，免疫表型，PD模型和修复的生化和行为测试，目标是开发一种新的治疗方案，用于保护和拯救人类PD中的高危神经元。使用来自脑或骨髓的自体细胞的可能性为开发干/祖细胞以及分化细胞以减缓甚至停止破坏性神经障碍的过程提供了巨大的治疗优势。据推测，星形细胞，无论是来自大脑或骨髓，有能力被改造释放治疗因子的PD。

DESCRIPTION (provided by applicant): There is a need to develop cell and molecular protection approaches for at-risk neurons in neurodegenerative diseases, and regenerative medicine offers designer cells to be used for factor-delivery in diseases like Parkinson's (PD) where we know that certain growth factors might, if delivered properly, have the ability to rescue a discrete population of nigrostriatal cells and hence slow or halt the progression of the disease. This proposal will use a novel cellular transplant approach to deliver glial cell line derived neurotrophic factor (GDNF) in comparable in vitro and in vivo bioassays of dopamine neuron degeneration. Three specific aims focus on which astrocytes, from different sources, can be used to deliver GDNF to at risk dopamine neurons in a culture model of PD as well as a well-accepted rodent model (6-hydroxydopamine, 6- OHDA lesions of the neostriatum) of the disease. Specific Aim 1 will establish the potential for autologous cellular repair by focusing on newly-generated "immature" astrocytes derived from the mouse brain (e.g. adult cerebral cortex and striatum), and bone marrow versus mouse embryonic stem cells. These candidate astrocyte populations will be transduced with an eGFP-GDNF lentiviral construct to determine which population(s) is most amenable to stable growth factor transduction and release of the growth factor, for subsequent testing in an in vivo lesion-protection model (Aim 3). Specific Aim 2 will look at adult human brain astrotypic and bone marrow-derived cells and their ability to be stably transduced with lenti-GDNF and release the growth factor as the mouse cells do in Aim 1. Specific Aim 3 will assay the potential protective actions of GDNF-releasing mouse and human cells in DA depleted mice. These studies will incorporate methods already developed and tested in our laboratories, including cell culture, electrophysiology, gene therapy, cell grafting, immunophenotyping, biochemical and behavioral testing at PD models and repair, with the goal of developing a new therapeutic protocol for at-risk neuron protection and rescue in human PD. The possibility of using autologous cells derived from brain or bone marrow offers a tremendous therapeutic advantage for exploiting stem/progenitor as well as differentiated cells to slow and even halt the course of devastating neurological disorders. It is hypothesized that astrotypic cells, derived either from brain or bone marrow, have the ability to be engineered to release therapeutic factors in PD.