Stromal Stem Cells for Therapy of the Gangliosidoses

描述（由申请人提供）： 该项目的首要目标是评估骨的应用 骨髓基质干细胞移植治疗神经节苷脂沉积症 使用人神经节苷脂沉积症的充分表征的真实猫模型。 我们将对猫骨髓进行全面的鉴定 基质细胞（MSC）在体外，重点是他们的能力，分化 中枢神经系统（CNS）的神经元和神经胶质。 我们已经分离出 和部分表征的猫骨髓基质细胞（MSC）， 形态学、体外生长性状、细胞表面抗原和 分化成神经元表型的细胞。 我们将继续 表征猫MSC以确定是否存在显著的种属差异 存在可能影响体内可塑性和功能的物质。 我们将移植猫MSC，分析其分布， 在中枢神经系统和非中枢神经系统器官的分化。 猫MSC 将被标记上受体基因，然后注射到正常的猫科动物体内， 受试者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 从这些细胞衍生的谱系。 MSC将被脑内注射， 脑室内或静脉内注射至8 - 12周龄猫或颅内 腹腔注射到猫胎中。 子宫内注射MSC的运输 或出生后将在大脑，骨髓和各种 实质器官 我们将正常或载体校正的MSC移植到患有GM1的猫中， 神经节苷脂沉积症评估治疗潜力。 治疗性移植 a）来源于父母或兄弟姐妹的正常MSC 供体将被标记一个报告基因，在子宫内注射到猫 胎仔，并在出生后评价标记细胞的命运和分布 和疾病的改善; B）从猫出生后收获的自体MSC GM1神经节苷脂沉积症的患者将用携带 溶酶体β-半乳糖苷酶cDNA以纠正其酶缺陷， 重新植入大脑或静脉注射。 小猫与 将评价神经节苷脂沉积的神经、肝脏和 胸腺疾病 干细胞移植可能改变 将研究神经节苷脂沉积的过程。

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.