Using Stem Cells in Animal Models of Parkinson's Disease

在帕金森病动物模型中使用干细胞

• 批准号: 6623107
• 负责人: LORRAINE IACOVITTI
• 金额: $ 33.21万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6623107
• 关键词: Parkinson's disease; bone marrow; cell cycle; disease /disorder model; dopamine; dopamine receptor; embryonic stem cell; high performance liquid chromatography; histogenesis; immunocytochemistry; laboratory rat; nervous system disorder therapy; neurogenesis; nonhuman therapy evaluation; northern blottings; polymerase chain reaction; stem cell transplantation; tissue /cell culture; tyrosine 3 monooxygenase

One promising new therapy for Parkinson's Disease (PD) involves the replacement of degenerated nigrostriatal neurons with those derived from transplanted fetal mesencephalic tissue. Although this approach has often yielded remarkable recovery of function in rats and monkeys, results in clinical trials with PD patients have been less consistent. At issue, is the relative inability to standardize a number of critical factors in human fetal transplants, including the age, type, number and integrity of cells being grafted. Consequently, finding more reliable sources of dopaminergic (DA) tissue for transplantation has become increasingly important. One direction has been to search for a line of readily available, well-characterized continually self-renewing stem or precursor cells that possess the capacity to differentiate, ideally spontaneously and with the need for little manipulation, into DA neurons, thus providing an inexhaustible and uniform source of replacement tissue. Towards this end, our preliminary findings demonstrate that grafts of embryonic mouse neural stem cells (NSCs) of the C17.2 cell can differentiate exclusively into neurons, which in a majority of cases, can express DA traits when cells are transplanted into the brain of a Parkinsonian rat. In addition, in preliminary studies using stem cells from adult human bone marrow (MSCs), we have found that nearly 100 percent of MSCs will convert into process- bearing, beta-tubulin III+ neuronal-like cells after only 1-2 hours of incubation with specific differentiation factors. If these cells also exhibit the same capacity as NSCs to respond to appropriate DA differentiation cues in vivo, patients could provide their own source of stem cells for autologous grafts in PD. Using NSC and MSC stem cell models and a multidisciplinary approach, our specific goals for this proposal are threefold: 1) Identify the conditions that promote the stable appearance of a postmitotic differentiated DA phenotype in stem cells grown in culture; 2) Identify those factors which promote the differentiation of a DA phenotype in transplanted stem cells and 3) Determine whether the DA phenotype in transplanted stem cells is stable and long lasting, and whether, it can produce functional recovery of motor deficits in a rat model of PD. The ultimate goal of this research program is a fuller understanding of the cellular and molecular processes regulating the differentiation of DA traits in stem cells and apply that knowledge to transplantation strategies for the treatment of Parkinson's Disease.

帕金森病 (PD) 的一种有前途的新疗法涉及用来自移植的胎儿中脑组织的神经元替换退化的黑质纹状体神经元。尽管这种方法常常使大鼠和猴子的功能得到显着恢复，但针对帕金森病患者的临床试验结果却不太一致。 问题在于相对无法标准化人类胎儿移植中的许多关键因素，包括移植细胞的年龄、类型、数量和完整性。 因此，寻找更可靠的多巴胺能（DA）组织来源用于移植变得越来越重要。 一个方向是寻找一系列易于获得、特征良好的持续自我更新的干细胞或前体细胞，这些细胞具有分化能力，理想情况下是自发的，并且需要很少的操作，分化成DA神经元，从而提供取之不尽、用之不竭的统一的替代组织来源。 为此，我们的初步研究结果表明，C17.2 细胞的胚胎小鼠神经干细胞 (NSC) 移植物可以专门分化为神经元，在大多数情况下，当细胞移植到帕金森病大鼠的大脑中时，可以表达 DA 特征。 此外，在使用成人骨髓 (MSC) 干细胞的初步研究中，我们发现近 100% 的 MSC 在与特定分化因子孵育仅 1-2 小时后就会转化为具有加工能力的 β-微管蛋白 III+ 神经元样细胞。 如果这些细胞也表现出与 NSC 相同的能力，能够在体内对适当的 DA 分化信号做出反应，那么患者就可以为 PD 中的自体移植提供自己的干细胞来源。使用 NSC 和 MSC 干细胞模型以及多学科方法，我们对该提案的具体目标有三个：1) 确定促进培养干细胞中有丝分裂后分化 DA 表型稳定出现的条件； 2) 确定促进移植干细胞中 DA 表型分化的因素，以及 3) 确定移植干细胞中 DA 表型是否稳定且持久，以及是否可以在 PD 大鼠模型中产生运动缺陷的功能恢复。 该研究项目的最终目标是更全面地了解调节干细胞 DA 特征分化的细胞和分子过程，并将这些知识应用于治疗帕金森病的移植策略。