PATHOPHYSIOLOGY AND TREATMENT OF HUMAN GENETIC DISEASES

我们继续进行研究，以阐明遗传性 结缔组织疾病的疾病，并将此信息应用于 治疗这些疾病。使用反义核糖核酸探针系统，我们有 检测到三个病例的I型胶原mRNA的错配， 成骨不全症（OI），现在正在从事分离突变体 用于测序的等位基因：一例COOH末端错配的致死性OI α 2（I），III型OI，α I（I）中间错配，和 IV型OI在α I（I）的5 '-末端和α I（I）的3'-末端具有错配， α 2（I）。 我们正在扩展我们的能力，以检测不匹配的实现， 化学切割方法和使用有义和反义 核糖核酸探针筛选PCR扩增的cDNA的两条链。我们正在申请 这些技术，特别是那些有证据表明 嵌合性、复合杂合性或表达变异性。 我们已经开始开发核糖探针系统和蛋白质分离 III型胶原蛋白的技术。检测系统的组合 I型和III型胶原蛋白的点突变旨在分子研究 Ehlers-Danlos（艾德）综合征患者。我们有 分析了五例患者培养的成纤维细胞合成的蛋白质， 艾德，并检测到I型胶原蛋白电泳异常 在三种情况下;这些情况下的mRNA现在正在筛选 核糖核酸探针技术 在临床方案中，我们继续（1）我们对 OI中的生长缺陷和OI骨对生长的反应性 刺激，和（2）物理康复和支撑协议， 中重度OI的儿童。我们还发起了一项合作 OI儿童骨钙动态变化的研究

We have continued our studies to elucidate the molecular basis of heritable disorders of Connective tissue disease and to apply this information to the treatment of these disorders. Using the antisense riboprobe system, we have detected mismatches in the type I collagen mRNA of three cases of osteogenesis imperfecta (OI) and are now engaged in isolating the mutant alleles for sequencing: a case of lethal OI with a mismatch in the COOH end of alpha 2(I), type III OI with a mismatch in the middle of alpha I(I), and type IV OI with mismatches in the 5'-end of alpha I(I) and the 3'-end of alpha 2(I). We are extending our ability to detect mismatches by the implementation of chemical cleavage methodology and by using both sense and antisense riboprobes to screen both strands of PCRamplified cDNA. We are applying these techniques especially to those cases in which there is evidence of mosaicism, compound heterozygosity or variability of expression. We have begun to develop a riboprobe system and protein isolation techniques for type III collagen. The combination of systems to detect point mutations in types I and III collagen is aimed at molecular studies of patients with various forms of Ehlers-Danlos (ED) syndrome. We have analyzed the protein synthesized by cultured fibroblasts in five cases of ED and have detected electrophoretic abnormalities in the type I collagen in three cases; the mRNA of these cases is now being screened by the riboprobe technology. In clinical protocols, we have continued (1) our investigation of the growth deficiency in OI and the responsiveness of OI bone to growth stimulation, and (2) the physical rehabilitation and bracing protocol for children with moderately severe OI. We have also initiated a collaborative investigation of the dynamics of skeletal calcium in OI children.