MPS VII CNS Gene Therapy Using Neuronal Stem Cells

描述（由申请人提供）：本提案的长期目标是开发一种治疗中枢神经系统（CNS）病理高度进行性溶酶体酶缺乏症的方法。治疗必须广泛适用，并导致永久稳定的治愈。粘多糖病VII型（MRS VII）小鼠缺乏溶酶体酶-葡萄糖醛酸酶（GUS），由于具有良好特征的中枢神经系统参与，因此非常适合这些目标。通过正常骨髓移植（BMT）或酶替代疗法（ERT）恢复GUS在发育早期对内脏器官和组织具有治疗作用。替代酶通过完成对积累/储存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贮积疾病（LSD），这种疾病在5000个活产婴儿中发生近1例。许多LSD的中枢神经系统有储存物质，对BMT和ERT反应不佳。中枢神经系统可移植神经干细胞（NSC）的发现为中枢神经系统的治疗带来了希望。障碍包括NSC来源/可用性和不相容细胞的免疫排斥。NSC可以从脐带血（CB）和骨髓（BM）中培养，提供比胎儿组织更少争议的来源。目的1验证了异位Noggin治疗在体外培养过程中提高小鼠成年BM和/或胎儿肝脏（相当于小鼠CB） NSC发育效率的假设。Noggin拮抗骨形态发生蛋白，导致神经元细胞与造血细胞发育增加。目的2验证共刺激阻断将建立宿主对移植异体中枢神经系统NSC的耐受性的假设。即使CB被证明是生成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反应。目的2将确定在有或没有通过阻断免疫细胞共刺激激活来成功实现同种异体骨髓移植的试剂时，宿主免疫排斥同种异体骨髓移植和胎肝来源的NSC的效率。目的3将确定正常骨髓和胎肝源性NSC以及用GUS cDNA慢病毒转导的MPS VII骨髓源性NSC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/可用性和免疫障碍。本提案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有关，不仅包括LSD，还包括帕金森病和阿尔茨海默病。

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.