MPS VII CNS Gene Therapy Using Neuronal Stem Cells

描述(由申请人提供)：这项提案的长期目标是开发一种治疗高度进行性溶酶体酶缺乏症的中枢神经系统(CNS)病理。治疗必须广泛适用，并导致永久稳定的治愈。粘多糖III型(MRS VII)小鼠缺乏溶酶体酶β-葡萄糖醛酸苷酶(GUS)，由于具有良好的中枢神经系统参与的特点，因此特别适合于这些目标。通过正常骨髓移植(BMT)或酶替代疗法(ERT)修复GUS对发育早期的内脏器官和组织具有治疗作用。被替换的酶通过完成对积累/储存的底物的消化来恢复细胞功能。这两种方法都广泛适用于溶酶体储存疾病(LSD)，这种疾病发生在近1/5,000个活产儿中。许多LSD的中枢神经系统具有对BMT和ERT反应不佳的储存材料。中枢神经系统可移植神经干细胞(NSC)的发现为治疗中枢神经系统疾病带来了希望。障碍包括NSC来源/可获得性和不相容细胞的免疫排斥。神经干细胞可以从脐带血(CB)和骨髓(BM)中培养出来，提供的来源比胎儿组织争议更少。目的1验证在体外培养过程中，异位Noggin处理可提高小鼠成年骨髓和/或胎肝(相当于小鼠的CB)的神经干细胞发育效率的假设。Noggin拮抗骨形态发生蛋白，导致神经细胞和造血细胞发育加快。目的2验证共刺激阻断将建立宿主对CNS移植的同种异体神经干细胞耐受性的假说。即使CB被证明是产生NSC的可靠来源，主要(或次要)组织相容性复合体抗原的差异也可能刺激免疫排斥。目的2将确定宿主免疫排斥异基因BM和胎肝来源的NSC的效率，当移植到脑内时，有和没有试剂，通过阻断免疫细胞的共刺激激活，使异基因骨髓移植成功。目的研究慢病毒转导GUS基因的正常骨髓、胎肝来源的神经干细胞和MPS VII来源的神经干细胞的治疗潜力。后一种方法模拟使用患者自己的骨髓，以避免NSC治疗的来源/可获得性和免疫障碍。这项建议的目的是解决成功治疗中枢神经系统疾病的几个重要障碍。所获得的知识将为成功替换大脑正常健康功能所需的缺失或缺陷细胞和/或蛋白质提供有价值的信息。发现绕过中枢神经系统细胞植入障碍的方法与进行性中枢神经系统疾病相关，这些疾病不仅包括LSD，还包括帕金森氏症和阿尔茨海默氏症。

DESCRIPTION (provided by applicant): The long-term goal of this proposal is to develop a treatment for the central nervous system (CNS) pathology of highly progressive lysosomal enzyme deficiencies. The treatment must be broadly applicable and lead to a permanent stable cure. The Mucopolysaccharidosis Type VII (MRS VII) mouse, lacking the lysosomal enzyme beta-glucuronidase (GUS), is uniquely suited to these goals because of the well-characterized CNS involvement. Restoration of GUS by normal bone marrow transplantation (BMT) or by enzyme replacement therapy (ERT) is therapeutic to visceral organs and tissues when performed early in development. The replaced enzyme restores cell function by completing digestion of accumulated/stored substrates. Both approaches are broadly applicable to lysosomal storage diseases (LSD) in general, which occur in nearly 1 in 5,000 live births. The CNS of many LSD have storage material that does not respond well to BMT and ERT. Discovery of CNS engraftable neuronal stem cells (NSC) which generate progeny that migrate to multiple regions of brain raises hope for treating the CNS. Obstacles include NSC source/availability and immune rejection of incompatible cells. NSC can be cultured from umbilical cord blood (CB) and bone marrow (BM), providing sources less controversial than fetal tissue. Aim 1 tests the hypothesis that ectopic Noggin treatment improves the efficiency of NSC development from murine adult BM and/or fetal liver (the murine equivalent of CB) during in vitro culture. Noggin antagonizes bone morphogenic proteins, causing increased neuronal versus hematopoietic cell development. Aim 2 tests the hypothesis that costimulatory blockade will establish host tolerance to CNS grafted allogeneic NSC. Even if CB proves to be a reliable source for generating NSC, differences in major (or minor) histocompatibility complex antigens could stimulate immune rejection. Aim 2 will determine how efficiently host immunity rejects allogeneic BM and fetal liver derived NSC when transplanted into brain with and without reagents that enable successful allogeneic BMT by blocking costimulatory activation of immune cells. Aim 3 will determine the therapeutic potential of normal BM and fetal liver derived NSC as well as MPS VII BM derived NSC lentivirally transduced with GUS cDNA. The latter approach simulates use of the patients own BM to avoid both source/availability and immunological barriers to NSC therapy. The aims of this proposal address several important barriers to successful treatment of diseases of the CNS. The knowledge gained will provided valuable information towards successful replacement of missing or defective cells and/or proteins needed by the brain for normal healthy function. Discovery of methods to circumvent barriers to CNS cellular engraftment is relevant to progressive CNS diseases that not only include LSD, but also Parkinson's and Alzheimer's diseases.