Therapeutic hair follicle-derived neurospheres

治疗性毛囊源性神经球

基本信息

批准号：
6934274
负责人：
MENG YANG
金额：
$ 14.98万
依托单位：
ANTICANCER, INC.
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：
2005-04-15 至 2006-04-14
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The ready availability of pluripotent adult stem cells would have many important therapeutic applications. Our recent results suggest that the hair follicle is a promising source for such stem cells. Our recent discovery that the neural stem cell marker, nestin is expressed in hair-follicle stem cells (Li, L., et al. Nestin expression in hair follicle sheath progenitor cells. Proc. Natl. Acad. Sci. USA 100, 9658-9662, 2003) suggested that hair-follicle stem cells and neural stem cells have common features. Recently, we have demonstrated that the hair follicle gives rise to blood vessels in vivo with the blood vessels apparently originating from the hair-follicle stem cells (Amoh, Y., et al. Nascent blood vessels in the skin arise from nestin-expressing hair follicle cells. Proc. Natl. Acad. Sci. USA, in press). In preliminary experiments, we have demonstrated in vitro that hair-follicle stem cells can be induced to form neurospheres, which in turn form neurons (Amoh, Y., Li, L., and Hoffman, R.M., unpublished results). These results suggest pluripotent hair follicle adult stem cells could have important therapeutic applications, in particular for neurological diseases. This application utilizes transgenic mice with green fluorescent protein (GFP) under the control of the nestin regulatory sequences as the source of labeled hair follicle cells. In the nestin-GFP transgenic mice, nestin-expressing hair-follicle stem cells are located in the permanent upper hair follicle right below the sebaceous glands and above the errector pili muscle in the follicle bulge area. This group of nestin-expressing cells in the bulge area of the follicle can be activated by anagen induction and form the new hair follicles as well as a blood-vessel network. Immunohistochemical staining revealed that the nestin-GFP-expressing cells in the hair follicle bulge co-express nestin, GFP, and keratin-15 one of the markers of hair-follicle stem cells. The blood vessels formed from the follicle co-express CD-31 an endothelial cell marker as well as GFP. We have shown that the isolated hair-follicle nestin-expressing cells can form neurospheres in vitro that can be switched to form either neurons or blood vessels. The specific aims of this application are as follows: (1) Optimization of isolation of nestin-GFP-expressing hair-follicle stem cells for neurosphere conversion; (2) Determination of the optimal conditions of hairfollicle- stem-cell-derived neurospheres to form neurons. In Phase II, we will use optimized hair follicle derived neorospheres developed in Phase I to determine their therapeutic efficiency to produce neurons, blood vessels, and hair follicles in vivo. Human hair-follicle stem cells will also be developed for therapeutic potential in Phase II.

描述（由申请人提供）：多能的成年干细胞的现成可用性将具有许多重要的治疗应用。我们最近的结果表明，毛囊是这种干细胞的有前途来源。我们最近发现的是，神经干细胞标记Nestin在毛囊干细胞中表达（Li，L。等，Nestl。在毛囊卵泡鞘祖细胞中表达。最近，我们证明了毛囊在体内产生血管，显然是由毛囊干细胞起源于毛囊干细胞的血管（Amoh，Y。等。皮肤中的新生血管发起了筑巢的脱粒毛囊细胞。在初步实验中，我们已经在体外证明了毛状干细胞可以诱导形成神经球，而神经球又形成了神经元（Amoh，Y.，Li，L。和Hoffman，R.M.，未发表的结果）。这些结果表明多能毛囊成年干细胞可能具有重要的治疗性应用，特别是对于神经系统疾病。该应用在Nestin调节序列的控制下利用具有绿色荧光蛋白（GFP）的转基因小鼠作为标记的毛囊细胞的来源。在Nestin-GFP转基因小鼠中，表达巢蛋白的毛囊干细胞位于皮脂腺下方的永久性上部毛囊中，卵泡凸出区域中的pil骨pilli肌肉上方。卵泡隆起区域中的这组表达巢蛋白的细胞可以通过Anagen诱导来激活，并形成新的毛囊以及血管网络。免疫组织化学染色表明，毛囊中的Nestin-GFP表达细胞共表达巢穴，GFP和角蛋白15毛状毛状干细胞的标记之一。由卵泡共表达CD-31形成的血管和GFP。我们已经表明，孤立的表达巢穴的细胞可以在体外形成神经球，可以切换成形成神经元或血管。该应用的具体目的如下：（1）优化表达巢蛋白-GFP的毛囊干细胞以进行神经圈转化的优化；（2）测定头部源自干细胞衍生的神经球的最佳条件以形成神经元。在第二阶段，我们将使用在I期中开发的优化毛囊衍生的新卵球体来确定它们在体内产生神经元，血管和毛囊的治疗效率。在II期中，还将开发人毛状干细胞的治疗潜力。