Cell Grafts for Parkinson's Disease

细胞移植治疗帕金森病

• 批准号: 6659863
• 负责人: Timothy J. Collier
• 金额: $ 36.25万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6659863
• 关键词: 6 hydroxydopamine; Parkinson's disease; basal ganglia; cell transplantation; cysteine endopeptidases; cytokine; dopamine; dopamine receptor; embryo /fetus tissue /cell culture; embryo /fetus tissue transplantation; enzyme activity; enzyme inhibitors; innervation; laboratory rat; mesencephalon; microdialysis; nerve growth factors; nervous system disorder therapy; nervous system transplantation; neurotoxicology; neurotrophic factors; nonhuman therapy evaluation; stem cell transplantation; succinate dehydrogenase; tyrosine 3 monooxygenase

DESCRIPTION (provided by applicant): In vitro expansion of neural progenitor cells followed by induction of dopaminergic phenotype may provide a limitless source of cells for grafting into patients with Parkinson's disease (PD). However, the signals controlling the conversion of these cells into dopamine (DA) neurons must be identified. In an effort to accomplish this, single cells isolated from ventral mesencephalon were clonally expanded and exposed to hematopoeitic cytokines and neurotrophic molecules. Analysis of cell differentiation in response to this treatment yielded conversion of a high percentage (72 to 98 percent) of cells in some clones to a tyrosine hydroxylase (TH)-positive phenotype. Of the 24 clones generated, the best conversion to TH cells occurred with exposure to a combination of interleukin-1 (IL-1), interleukin- 11 (IL-11), leukemia inhibitory factor (LIF), and glial cell line-derived neurotrophic factor (GDNF). Positive clones expressed TH, the DA transporter, Nurr-1 and released DA in culture. Other cells in cytokine-exposed clones expressed GFAP (astrocyte marker) or MAP-2 (neuron marker) indicating that the original neurospheres were also capable of producing clones that differentiate into glial and nondopaminergic neurons. Initial neural grafting studies m the rat model of PD using a clone with the highest conversion rate to TH indicated that converted progenitor cell grafts produced complete amelioration of amphetamine-induced rotational behavior and continued to express the TH phenotype. However, the survival rate of these grafted progenitor cells was reduced (26 percent) compared to embryonic ventral mesencephalon (VM). The experiments proposed here will develop protocols for optimal survival of Wafted cytokine-converted mesencephalic progenitor cells. Once survival of grafted mesencephalic progenitor cells is optimized, direct comparisons will be made to fresh embryonic VM grafts on measures of behavior, in vivo dialysis, post-mortem DA biochemistry, DA receptors, cell survival and neurite extension. Lastly, this proposal will test the efficacy of the DA conversion cocktail on clonal progenitors derived from embryonic mesencephalon of nonhuman primate brain. If successful, cytokine-converted mesencephalic progenitor cells could potentially replace embryonic tissue as the primary source of cells for grafting in PD.

描述（由申请人提供）：神经祖细胞的体外扩张 细胞随后诱导多巴胺能表型可能会提供无限的 嫁接帕金森氏病（PD）患者的细胞来源。 但是，控制这些细胞转化为多巴胺的信号 （DA）必须识别神经元。为了实现这一目标，单细胞 从腹室中分离出来，将克隆膨胀并暴露于 造血细胞因子和神经营养分子。细胞分析 响应于这种治疗的分化产生了高的转化 在某些克隆到酪氨酸羟化酶的细胞百分比（72％至98％） （Th） - 阳性表型。在产生的24个克隆中，最好的转换为 细胞发生在暴露于白介素-1（IL-1）的组合中， 白介素11（IL-11），白血病抑制因子（LIF）和神经胶质细胞 线来衍生的神经营养因子（GDNF）。积极克隆表达了TH，DA 转运蛋白，Nurr-1并在文化中发布DA。细胞因子暴露的其他细胞 克隆表达GFAP（星形胶质细胞标记）或MAP-2（神经元标记），指示 原始的神经球也能够产生克隆 分化为神经胶质和非阿顿胺能神经元。最初的神经嫁接 使用最高转化率至 这表明产生的转化后祖细胞移植已完成 改善苯丙胺引起的旋转行为，并继续 表达表型。但是，这些移植的存活率 与胚胎腹侧相比，祖细胞降低（26％） 中脑（VM）。这里提出的实验将为 伴有细胞因子转换的中脑祖细胞的最佳存活。 一旦优化了移植的中脑祖细胞的存活，直接 将与新鲜胚胎VM移植物进行比较，以进行行为度量， 体内透析，后DA生物化学，DA受体，细胞存活和 神经突延伸。最后，该建议将测试DA的功效 源自胚胎中脑的克隆祖细胞上的转换鸡尾酒 非人类灵长类动物的大脑。如果成功，细胞因子转换的中脑 祖细胞可能会替代胚胎组织 在PD中嫁接的细胞来源。