STEM/PROGENITOR CELL PROTECTION FOR PARKINSON'S DISEASE

描述（由申请人提供）：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，再生医学提供了用于帕金森病（PD）等疾病的因子递送的设计细胞，我们知道某些生长因子如果递送得当，可能有能力拯救离散的黑质纹状体细胞群，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，在多巴胺神经元变性的可比体外和体内生物测定中传递胶质细胞系衍生神经营养因子（GDNF）。三个特定的目标集中在哪些来自不同来源的星形胶质细胞可以用于在PD培养模型中以及被广泛接受的啮齿动物模型（6-羟多巴胺，6- OHDA病变的新纹状体）中将GDNF传递到危险的多巴胺神经元。特异性目标1将通过关注来自小鼠大脑（如成人大脑皮层和纹状体）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的对比，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慢病毒构建体进行转导，以确定哪个星形胶质细胞群最适合稳定的生长因子转导和生长因子的释放，以便随后在体内损伤保护模型中进行测试（目标3）。特异性目标2将研究成人大脑星形型细胞和骨髓来源细胞，以及它们稳定地用lentig - gdnf转导和释放生长因子的能力，就像目标1中的小鼠细胞一样。特异性Aim 3将检测gdnf释放小鼠和人细胞对DA缺失小鼠的潜在保护作用。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，用于高危神经元保护和拯救人类PD。使用来自大脑或骨髓的自体细胞的可能性为利用干细胞/祖细胞以及分化细胞来减缓甚至停止破坏性神经系统疾病的进程提供了巨大的治疗优势。据推测，来自大脑或骨髓的星形型细胞具有被改造以释放PD治疗因子的能力。

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.