Correction of Hurler syndrome by multipotent stem cells

多能干细胞纠正Hurler综合征

• 批准号: 7097548
• 负责人: Pankaj GUPTA
• 金额: $ 2.37万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7097548
• 关键词: SCID mouse; biosynthesis; cell surface receptors; cytokine; enzyme activity; extracellular matrix; gene expression; human subject; immunoprecipitation; laboratory mouse; microarray technology; mucopolysaccharides; mucopolysaccharidosis type I; neuropathology; neuropsychology; oligosaccharides; patient oriented research; pluripotent stem cells; stem cell transplantation; tissue /cell preparation; western blottings

DESCRIPTION (provided by applicant): Novel approaches based on pathophysiologic mechanisms are needed to treat progressive neurological dysfunction and mental retardation in Hurler syndrome (MPS I). Our group has identified a multipotent adult progenitor cell (MAPC) in human and rodent bone marrow (BM), which differentiates in vitro and in vivo into all three embryonic lineages, including neuronal and glial cells (Blood 98:2615, 2001; Nature 418:41, 2002). MAPC implanted into adult rodent brains engraft, acquire neuronal and glial markers and ameliorate ischemic neurological dysfunction. MAPC may be ideal for neural tissue reconstitution/gene delivery, as they 1) are obtained from a small BM sample and proliferate without senescence 2) are efficiently transduced and express genes without losing stem cell potential 3) are free from ethical issues of embryonic/fetal tissues. During an R03 grant, we found that 1) structurally abnormal heparan sulfates (HS) in Hurler MAPC have defective binding interactions with FGF-2, thus impairing its biological activity on MAPC survival, proliferation and neural differentiation 2) normal MAPC provide trophic and corrective signals to Hurler MAPC 3) gene expression of proteoglycans (PGs), glycosaminoglycan (GAG) metabolic enzymes, cytokines and morphogens is altered in Hurler MAPC. Separately, our group found severe neuropathological and behavioral abnormalities in a murine knockout model of MPS I. We hypothesize that 1) the structural and functional abnormalities of HS contribute to the neuronal pathophysiology of Hurler syndrome by perturbing the biological activity of critical cytokines and by secondarily modulating gene expression, and 2) following intra-ventricular transplantation, normal human MAPC will engraft in the brain of neonatal immunodeficient (NOD-SCID)-MPS I mice and ameliorate behavioral abnormalities. Specific Aim (SA) 1: Compare the structure and function of HS from normal and Hurler MAPC and murine brain tissue. SA2: Test the secondary effects of accumulated GAGs and oligosaccharides on HS biosynthesis and gene expression in Hurler MAPC. SA3: Examine if the neuropathology and behavioral dysfunction in NOD-SCID-MPS I mice can be corrected by intracerebroventricular transplantation of normal human MAPC. If MAPC transplantation is of benefit in the Hurler model, similar strategies may be developed for treating diverse neurodegenerative disorders.

描述（由申请人提供）：需要基于病理生理机制的新方法来治疗 Hurler 综合征（MPS I）中的进行性神经功能障碍和精神发育迟滞。我们的小组在人类和啮齿动物骨髓 (BM) 中鉴定出了一种多能成体祖细胞 (MAPC)，它在体外和体内分化为所有三种胚胎谱系，包括神经元和神经胶质细胞 (Blood 98:2615, 2001; Nature 418) ：41，2002）。 将 MAPC 植入成年啮齿动物大脑中，获得神经元和神经胶质标记物并改善缺血性神经功能障碍。 MAPC 可能是神经组织重建/基因传递的理想选择，因为它们 1) 从小 BM 样本中获得，并且增殖而不衰老 2) 有效转导和表达基因，而不会失去干细胞潜力 3) 不存在胚胎/细胞的伦理问题胎儿组织。在 R03 资助期间，我们发现 1) Hurler MAPC 中结构异常的硫酸乙酰肝素 (HS) 与 FGF-2 的结合相互作用有缺陷，从而损害了其对 MAPC 存活、增殖和神经分化的生物活性 2) 正常 MAPC 提供营养和纠正作用向 Hurler MAPC 发出信号 3) 蛋白聚糖 (PG)、糖胺聚糖 (GAG) 代谢酶、细胞因子和形态发生素的基因表达发生改变在 Hurler MAPC 中。另外，我们的小组在 MPS I 小鼠敲除模型中发现了严重的神经病理学和行为异常。我们假设 1) HS 的结构和功能异常通过扰乱关键细胞因子的生物活性以及其次通过调节基因表达，2) 脑室内移植后，正常人 MAPC 将植入新生免疫缺陷 (NOD-SCID)-MPS I 小鼠的大脑中，改善行为异常。具体目标 (SA) 1：比较正常和 Hurler MAPC 以及小鼠脑组织的 HS 的结构和功能。 SA2：测试累积的 GAG 和寡糖对 Hurler MAPC 中 HS 生物合成和基因表达的二次影响。 SA3：检查 NOD-SCID-MPS I 小鼠的神经病理学和行为功能障碍是否可以通过正常人 MAPC 的脑室内移植来纠正。如果 MAPC 移植在 Hurler 模型中有益，那么可以开发类似的策略来治疗不同的神经退行性疾病。