Krabbe Disease-Molecular Analysis and Treatment

克拉伯病-分子分析与治疗

• 批准号: 6711795
• 负责人: DAVID A WENGER
• 金额: $ 35.52万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6711795
• 关键词: Krabbe' s disease; O glycosidase; cell proliferation; disease /disorder model; dogs; enzyme activity; enzyme biosynthesis; gene expression; gene mutation; gene therapy; genetic regulatory element; hematopoietic tissue transplantation; insulinlike growth factor; laboratory mouse; molecular pathology; myelin basic proteins; myelination; neurogenetics; nonhuman therapy evaluation; oligodendroglia; sphingosine; stem cell transplantation; tissue /cell culture; transfection /expression vector

DESCRIPTION (provided by applicant): Krabbe disease or globoid cell leukodystrophy is a severe disorder of CNS and PNS myelin caused by mutations in the gene coding for galactocerebrosidase (GALC). The accumulation of one substrate, psychosine, is cytotoxic and induces the apoptotic death of oligodendrocytes (OLG) in the CNS and Schwann cells in the PNS. This is a project to treat murine and canine models using several approaches, including gene therapy and stem cell transplantation. A new transgenic mouse model (trs5O2) with low GALC activity was generated and will be utilized in some treatment studies. These mice live longer than twitcher (twi) mice and have large litters resulting in more affected pups for study. Recent studies show that the human GALC is not efficiently processed in rodent tissues so mouse GALC (mGALC) cDNA will be cloned into several viral vectors, including MFG, AAV and avian C-type retrovirus (SNV) for in vivo and ex vivo gene therapy trials. Insulin-like growth factor-I (IGF-1) has been shown to induce the proliferation of OLG progenitors, to increase remyelination after chemical or viral insults, and to prevent apoptotic death of OLG. Therefore, successful treatment of this disease may require delivery of GALC activity to prevent psychosine accumulation plus IGF-I to prevent apoptotic cell death and to induce remyelination. Initially IGF-1 will be injected subcutaneously in trsSO2 and twi mice to evaluate its effect clinically, biochemically and pathologically. A bi-cistronic viral vector containing mGALC cDNA plus IGF-I cDNA will be constructed in order to supply GALC activity plus IGF-1 to induce remyelination. A transgenic mouse that expresses IGF-1 under control of the myelin basic promoter will be generated and mated with GALC deficient mice to investigate its role in inducing remyelination when GALC activity is supplied via gene therapy or stem cell transplantation. Transduced and untransduced hematopoietic stem cells will be transplanted in the mouse and canine models with IGF-l supplementation. The effect of L-cycloserine to prolong life of the munne and canine models will be investigated. Treated dogs will be followed clinically and monitored by MRI and biochemical and pathological studies.

描述(申请人提供)：克拉贝病或球状细胞 脑白质营养不良是由突变引起的一种严重的中枢神经系统和三叉神经节髓鞘疾病 在编码半乳糖脑苷酶(GALC)的基因中。一个人的积累 底物，精神病素，具有细胞毒性，并诱导细胞凋亡死亡 中枢神经系统有少突胶质细胞，三叉神经节有雪旺细胞。这是一个 使用几种方法治疗小鼠和犬类模型的项目，包括 基因治疗和干细胞移植。一种新的转基因小鼠模型 产生了具有低GALC活性的(Trs5O2)，并将在一些 治疗研究。这些小鼠比Twitcher(双胞胎)小鼠活得更长，而且 大窝导致更多受影响的幼崽用于学习。最近的研究表明 人类的GALC在啮齿动物组织中不能有效地处理，所以小鼠 GALC(MGALC)基因将被克隆到包括MFG、AAV在内的几种病毒载体中 以及用于体内和体外基因治疗试验的禽C型逆转录病毒(SNV)。 胰岛素样生长因子-1(IGF-1)已被证明可诱导细胞增殖。 OLG前体细胞，在化学或病毒侮辱后增加重新髓鞘形成， 预防OLG的细胞凋亡性死亡。因此，成功的治疗这一点 疾病可能需要提供GALC活动以防止精神紧张 积聚加IGF-I预防细胞凋亡和诱导 重新髓鞘形成。最初，胰岛素样生长因子-1将被注射到trsSO2和 对TWI小鼠进行临床、生化和病理等方面的评价。一个 含有mGALC和IGF-I基因的双顺反子病毒载体将成为 构建用于提供GALC活性和IGF-1以诱导 重新髓鞘形成。在基因控制下表达IGF-1的转基因小鼠 髓鞘碱性启动子将被产生并与GALC缺陷小鼠交配 研究其在提供GALC活性时诱导再髓鞘形成中的作用 通过基因治疗或干细胞移植。换能和非换能 造血干细胞将被移植到小鼠和狗的模型中 补充胰岛素样生长因子-L。L-环丝氨酸对小鼠的延寿作用 将对孟尼和犬类模型进行研究。接受治疗的狗将被跟踪 经临床、核磁共振、生化及病理检查监测。