ANIMAL MODELS OF MUCOPOLYSACCHARIDOSES

粘多糖动物模型

• 批准号: 3227589
• 负责人: MARK E HASKINS
• 金额: $ 28.39万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3227589
• 关键词: animal colony; antibody; autologous transplantation; biopsy; bone marrow transplantation; cats; chromatography; complementary DNA; disease /disorder model; dogs; electrofocusing; electron microscopy; enzyme mechanism; enzyme therapy; fibroblasts; gene therapy; genetic library; genetic manipulation; histopathology; immunoelectrophoresis; molecular cloning; molecular pathology; mucopolysaccharidosis; mucopolysaccharidosis type I; mucopolysaccharidosis type VI; northern blottings; oligonucleotides; orphan disease /drug; tissue /cell culture

The broad objective of the proposed research is to investigate the pathogenesis of selected animal models of human mucopolysaccharidoses (MPS) and use them to develop and evaluate various therapeutic strategies. Inherent in these studies is the characterization of the molecular pathology of each animal model to determine the degree of homology to the respective human disease at the clinical, pathologic, biochemical, and genetic levels. Since the studies necessary cannot always be done in human patients, these models are an important source of knowledge. The analogues to be studied include: MPS I (Hurler syndrome, alpha-L- iduronidase deficiency), MPS VI (Maroteaux-Lamy syndrome, arylsulfatase B deficiency), and MPS VII (Sly disease, beta- glucuronidase deficiency). In addition, a new model will be created, a double homozygote MPS I/VI (alpha-L- iduronidase/arylusulfase B deficiency), that will allow new investigations into pathogenesis. The specific aims include: (1) The characterization of the natural history and pathologic manifestations of the animal analogues, in particular the disease of the brain and spinal cord, skeleton, heart, white blood cells, pigment epithelium, and liver; (2) Characterizing the biochemical defects in the MPS homologues will be done by purifying and determining the physicokinetic properties of the normal and abnormal enzymes, developing monoclonal antibodies, and measuring the glycosaminoglycan substrates; (3) Isolating cDNA and/or genomic clones encoding the normal enzymes using synthesized oligonucleotide probes and/or antibodies, collinear sequencing, and cDNA to probe genomic libraries; (4) Characterizing the molecular lesions in the MPS homologues by northern blotting, RNase analysis, and isolating, sequencing, and identifying the MPS mutations; (5) Developing and evaluating therapeutic modalities including: (a) enzyme manipulation therapy using compounds that increase residual enzyme activity in vitro and in vivo; (b) cellular manipulation therapy using bone marrow transplantation; (c) gene therapy using retroviral vectors developed to transfer the appropriate normal cloned gene into cultured fibroblasts, retinal pigment epithelium, and bone marrow cells prior to in vivo transplantation of transfection autologous bone marrow stem cells; (d) evaluate the therapy clinically, biochemically, and pathologically to determine the safety and efficacy of the procedures.

拟议研究的广泛目标是调查 人类选定动物模型的发病机理 粘二糖（MPS）并使用它们来开发和评估 各种治疗策略。 这些研究固有的是 每个动物模型的分子病理学的表征 确定与各自的人类疾病的同源性程度 在临床，病理，生化和遗传水平上。 由于必要的研究不能总是在人类中进行 患者，这些模型是重要的知识来源。 这 要研究的类似物包括：MPS I（Hurler综合征，alpha-l-- IDuronidase缺乏症），MPS VI（Maroteaux-Lamy综合征， Arylsulfatase B缺乏症）和MPS VII（狡猾的疾病，β- 葡萄糖醛酸酶缺乏症）。 此外，新模型将是 创建的是双纯合MPS I/VI（alpha-l-） iduronidase/arylususulfase b缺乏症），这将允许新 对发病机理的研究。 具体目的包括：（1）自然的表征 动物类似物的历史和病理表​​现， 特别是大脑和脊髓疾病，骨骼， 心脏，白细胞，色素上皮和肝脏； （2） 表征MPS同源物中的生化缺陷将 通过纯化和确定物理动力学特性来完成 正常酶和异常酶的产生单克隆 抗体，并测量糖胺聚糖底物； （3） 隔离编码正常酶的cDNA和/或基因组克隆 使用合成的寡核苷酸探针和/或抗体， 分线测序和cDNA以探测基因组文库； （4） 表征MPS同源物中的分子病变 北印迹，RNase分析和隔离，测序和 识别MPS突变； （5）开发和评估 治疗方式包括：（a）酶操纵疗法 使用可以在体外增加残留酶活性的化合物 体内; （b）使用骨髓的细胞操纵疗法 移植； （c）使用逆转录病毒载体的基因治疗 开发以将适当的正常克隆基因转移到 培养的成纤维细胞，视网膜色素上皮和骨髓 细胞在体内移植自体内自体移植 骨髓干细胞； （d）临床评估该治疗， 从生化和病理上，以确定安全性和 程序的功效。