ES Cell-Derived Motoneurons from SMA transgenic mice

来自 SMA 转基因小鼠的 ES 细胞衍生的运动神经元

基本信息

批准号：
6873080
负责人：
DOUGLAS A KERR
金额：
$ 37.61万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-04-01 至 2009-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): We propose a strategy that relies upon the differentiation of embryonic stem (ES) cells to motoneurons in order to define molecular and cellular abnormalities in Spinal Muscular Atrophy (SMA). We have generated ES cell lines from SMA transgenic mice and have shown that these lines undergo axonal degeneration and ultimately cellular death when differentiated into motoneurons in the presence of skeletal muscle. In Specific Aim 1, we will rigorously define the ability of SMA ES cell-derived motoneurons to form functional neuromuscular junctions with co-cultured skeletal muscle. We will also define abnormalities in axonal transport and the activity of axonal- or cell-death pathways. Specific Aim 2 will define the importance of SMA skeletal muscle in the observed motoneuron phenotype by carrying out selected experiments from Specific Aim 1 with SMA skeletal muscle in co-culture with normal ES cell-derived motoneurons and with SMA ES cell-derived motoneurons. Specific Aim 3 will determine the spatial requirements of SMN to rescue the abnormal motoneuron phenotype by transfecting SMN derivatives that differentially localize to the nucleus, cytoplasm and axons. Specific Aim 4 will define the temporal requirements of SMN to rescue the motoneuron phenotype by utilizing pharmacologic therapies that modulate exon-7 inclusion from SMN2 at various stages during differentiation. Specific Aim 5 will define the phenotype of SMA ES cell-derived motoneurons in ovo following transplantation into embryonic chick spinal cord, thereby defining an abnormality of axon extension or pathfinding. This project will identify critical molecular clues to motoneuron dysfunction in SMA and will identify potential strategies to halt or reverse this dysfunction.

描述（由申请人提供）：我们提出了一种依赖于胚胎干（ES）细胞分化为运动神经元的策略，以定义脊髓性肌萎缩症（SMA）中的分子和细胞异常。我们从 SMA 转基因小鼠中产生了 ES 细胞系，并表明这些细胞系在骨骼肌存在下分化为运动神经元时会发生轴突变性并最终发生细胞死亡。在具体目标 1 中，我们将严格定义 SMA ES 细胞衍生的运动神经元与共培养的骨骼肌形成功能性神经肌肉接头的能力。我们还将定义轴突运输的异常以及轴突或细胞死亡途径的活动。具体目标 2 将通过使用 SMA 骨骼肌与正常 ES 细胞衍生的运动神经元和 SMA ES 细胞衍生的运动神经元共培养进行特定目标 1 中选定的实验，来定义 SMA 骨骼肌在观察到的运动神经元表型中的重要性。具体目标 3 将确定 SMN 的空间要求，通过转染差异定位于细胞核、细胞质和轴突的 SMN 衍生物来挽救异常运动神经元表型。具体目标 4 将定义 SMN 的时间要求，通过利用药物疗法在分化过程的各个阶段调节 SMN2 的外显子 7 包含来挽救运动神经元表型。具体目标 5 将定义移植到胚胎鸡脊髓后卵内 SMA ES 细胞衍生的运动神经元的表型，从而定义轴突延伸或寻路的异常。该项目将确定 SMA 运动神经元功能障碍的关键分子线索，并将确定阻止或逆转这种功能障碍的潜在策略。