STEM/PROGENITOR CELL PROTECTION FOR PARKINSON'S DISEASE

描述(申请人提供)：需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，再生医学提供设计细胞用于帕金森病(PD)等疾病的因子传递，我们知道如果适当传递某些生长因子，可能有能力拯救离散的黑质纹状体细胞群，从而减缓或阻止疾病的进展。这项建议将使用一种新的细胞移植方法，在多巴胺神经元变性的体外和体内生物检测中传递神经胶质细胞系衍生神经营养因子(GDNF)。三个特定的目标集中在来自不同来源的星形胶质细胞可以用来向帕金森病的培养模型和被广泛接受的啮齿动物模型(6-羟基多巴胺，新纹状体的6-OHDA损伤)中的高危多巴胺神经元输送GDNF。具体目标1将通过关注来自小鼠大脑(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，建立自体细胞修复的可能性。这些候选星形胶质细胞群体将被绿色荧光蛋白-胶质细胞源性神经营养因子慢病毒载体转导，以确定哪个群体(S)最容易接受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(AIM 3)。特定目标2将观察成人大脑星形细胞和骨髓来源的细胞，以及它们被Lenti-GDNF稳定转导并释放生长因子的能力，就像目标1中的小鼠细胞一样。特定目标3将分析释放GDNF的小鼠和人类细胞对DA耗竭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、帕金森病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，用于人类帕金森病高危神经元的保护和拯救。使用来自大脑或骨髓的自体细胞的可能性为利用干细胞/祖细胞和分化细胞减缓甚至阻止破坏性神经疾病的进程提供了巨大的治疗优势。假设来自脑或骨髓的星形细胞具有在帕金森病中释放治疗因子的工程能力。

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.