Stromal Stem Cells for Therapy of the Gangliosidoses

描述(由申请人提供)： 本项目的总体目标是评估骨的应用。 骨髓基质干细胞移植治疗神经节细胞增多症 使用具有良好特征的、真实的人类神经节苷脂增多症的猫模型。 我们将对猫的骨髓进行全面的表征 基质细胞(MSC)的体外培养，重点是其分化能力 至中枢神经系统(CNS)的神经元和胶质细胞。我们已经隔离了 和部分特征化的猫骨髓基质细胞(MSC)基于 形态，体外生长特性，细胞表面抗原和能力 分化为神经表型的细胞。我们将继续 确定猫科动物间充质干细胞是否存在显著物种差异的特征 存在可能影响体内可塑性和功能的物质。 我们将移植猫的MSC来分析它们的分布和 中枢神经系统和非中枢神经系统器官的分化。猫科MSC 会被标记上受体基因，然后注射到正常的猫科动物体内 并在不同的时间点进行分析，以评估神经元和神经胶质细胞 从这些细胞衍生而来的血统。MSC将被注射到大脑内， 8-12周龄猫脑室或静脉注射或脑内注射 并通过腹膜进入猫胎体内。子宫内注射骨髓间充质干细胞的贩运 或出生后将在大脑、骨髓和各种 实质器官。 我们将把正常的或载体矫正的MSC移植到感染GM1的猫体内 评估神经节苷脂沉着症的治疗潜力。治疗性移植 猫的MSC将包括：a)来自父母或兄弟姐妹的正常MSC 捐赠者将被标记有报告基因，在子宫内注射到猫体内 胎儿，并在出生后评估标记细胞的命运和分布 和疾病的改善；b)出生后从猫身上获得的自体骨髓间充质干细胞 在GM1神经节苷脂中毒的情况下，将用携带有 溶酶体β-半乳糖苷酶基因纠正其酶缺陷和 重新植入大脑或静脉注射。小猫和小猫 将评估神经节苷脂增多症的神经、肝脏和 胸腺疾病。干细胞移植可能改变的机制 神经节苷脂增多症的病程将被研究。

DESCRIPTION (provided by applicant): The overarching goal of this project is to evaluate the application of bone marrow stromal stem cell transplantation for therapy of the gangliosidoses using well characterized, authentic feline models of the human gangliosidoses. We will undertake a comprehensive characterization of feline bone marrow stromal cells (MSC) in vitro, with emphasis on their capacity to differentiate to neurons and glial of the central nervous system (CNS). We have isolated and partially characterized feline marrow stromal cells (MSC) based on morphology, in vitro growth traits, cell surface antigens and capacity to differentiate to cells of a neuronal phenotype. We will continue characterization of feline MSC to determine if significant species differences exist which may influence plasticity and function in vivo. We will transplant feline MSC to analyze their distribution and differentiation in the central nervous system and non-CNS organs. Feline MSC will be labeled with a receptor gene, then injected into normal feline subjects and analyzed at various time points to assess the neuronal and glial lineages derived from such cells. MSC will be injected intra cerebrally, intraventricularly or intravenously into 8-12 week old cats or intracranially and intra peritoneally into cat fetuses. Trafficking of MSC injected in utero or postnatally will be monitored in brain, bone marrow and a variety of parenchymatous organs. We will transplant normal or vector corrected MSC into cats affected with GM1 gangliosidosis to assess therapeutic potential. Therapeutic transplantation of feline MSC will include: a) Normal MSC derived from parent or sibling donors will be labeled with a reporter gene, injected in utero into cat fetuses, and evaluated after birth for fate and distribution of labeled cells and amelioration of disease; b) Autologous MSC harvested postnatally from cats with GM1 gangliosidosis will be transfected with a lentiviral vector bearing a lysosomal beta-galactosidase cDNA to correct their enzymatic deficiency and re-implanted into brain or injected intravenously. Kittens with the gangliosidoses will be evaluated for progression of neurological, hepatic and thymic disease. Mechanisms by which stem cell transplantation may alter the course of the gangliosidoses will be studied.