HERITABLE DISORDERS OF CONNECTIVE TISSUE

遗传性结缔组织疾病

• 批准号: 6162427
• 负责人: J C MARINI
• 金额: --
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6162427
• 关键词: Ehlers Danlos syndrome; antisense nucleic acid; bone density; child (0-11); child physical development; clinical research; collagen; connective tissue metabolism; disease /disorder model; gene mutation; gene therapy; genetic disorder; genetic mapping; hormone therapy; human genetic material tag; human subject; laboratory mouse; molecular pathology; osteoblasts; osteogenesis imperfecta; rehabilitation; ribozymes; somatotropin; tissue /cell culture; transfection /expression vector

The Section has continued its studies aimed at elucidating the molecular mechanisms of heritable disorders of connective tissue, specifically osteogenesis imperfecta (OI) and Ehlers-Danlos (EDS), and at applying this information to the treatment of these disorders. One continuing interest of the Section is to identify the collagen mutations in patients with OI and EDS and determine the relationship between the type and location of the mutation and the severity of the connective tissue disorder. Mutations in the alpha2(I) collagen chain identified by this Section and other labs have provided additional support for the regional model we have proposed. There are three primary projects in the Section. One primary project is to develop selective antisense suppression of the mutant collagen allele as an approach for therapeutic intervention. We have been using hammerhead ribozymes as our selective agent. We are engaged in transferring our in vitro success with ribozymes into cultured fibroblasts. We are currently engaged in making stably transfected lines with known collagen mutations and in a transient transfection reporter system. The second primary project is the generation of a murine model for non-lethal OI. We have used site-directed mutagenesis to produce a construct with a point mutation mimicking non-lethal human OI. To make the construct conditional a lox-stop-lox construct has been introduced into the intron prior to the exon containing the disease-causing mutation. F1 mice have been generated from crosses with both wild type and cre-recombinase expressing mice. Recombination at the lox sites generates mutant protein expression and a severe OI skeletal phenotype. A third major focus of interest which we have been developing is in the bone biology of OI. We are using cultured osteoblasts to study the way bone cells modify and secrete mutant collagen. We are also pursing the secondary non-collagenous abnormalities of OI matrix and the response of OI osteoblasts to TGF-beta stimulation. In clinical studies, we are continuing our treatment trial of growth hormone in short children with OI to determine its effects on growth stimulation, bone density and bone morphometric properties. We are continuing our collaborative interests in the neurological aspects of OI and in maximizing the physical functioning of OI children though aggressive rehabilitation.

该科继续进行研究， 结缔组织遗传性疾病的机制，特别是 成骨诱导因子（OI）和Ehlers-Danlos（EDS）， 这些信息对这些疾病的治疗。 该科的一个持续兴趣是鉴定胶原蛋白， OI和EDS患者中的突变，并确定 突变的类型和位置与 结缔组织疾病α 2（I）胶原蛋白链突变 本节和其他实验室确定的其他实验提供了 支持我们提出的区域模式。 该科有三个主要项目。一个主要项目是 开发突变胶原等位基因的选择性反义抑制， 作为一种治疗干预的方法。我们一直在使用 锤头状核酶作为我们的选择剂我们从事 将我们在体外成功的核酶转移到培养的 成纤维细胞我们目前正致力于建立稳定转染的细胞系 具有已知的胶原蛋白突变和瞬时转染报告基因 系统 第二个主要项目是产生一个小鼠模型， 非致命性OI我们已经使用定点突变来产生一种 具有模拟非致死性人OI的点突变的构建体。使 已经引入了构造条件lox-stop-lox构造 插入内含子，位于含有致病基因的外显子之前 突变F1小鼠由与两种野生型 和表达cre重组酶的小鼠。液氧场的蒸发 产生突变蛋白质表达和严重的OI骨骼表型。 我们一直在发展的第三个主要关注点是 骨生物学我们用培养的成骨细胞来研究 骨细胞修饰并分泌突变胶原。我们也在追求 OI基质的继发性非胶原异常和 OI成骨细胞对TGF-β刺激的反应。 在临床研究中，我们正在继续我们的治疗试验的增长 激素对发育不全矮小儿童生长影响 刺激、骨密度和骨形态测量特性。我们 继续我们在OI神经方面的合作兴趣 最大限度地发挥OI儿童的身体功能， 积极的康复