ABNORMAL HEMOGLOBIN SYNTHESIS--MECHANISMS AND DETECTION

描述：本提案是对正常的研究的继续 镰状细胞性贫血与血红蛋白病的异常合成 地中海贫血。这些遗传性贫血的影响范围很广。 地理区域，包括地中海地区、非洲、中东 东亚、印度次大陆和东南亚。研究将会 专注于三个不同的领域，这将有助于 改善对这些疾病的管理。1)过去的研究资助 多年的研究表明，反向网点印迹是最简单的 检测镰状细胞性贫血突变的非放射性方法。 因此，在未来几年中，分析将被设计为涵盖大多数 在不同种族和地理区域的突变 以加快产前诊断。另外，因为胎儿细胞 已知在怀孕早期进入母体循环，研究 将通过简单的方法将胎儿细胞从 母亲S的血液，以便提供一种非侵入性的胎儿DNA 检测镰状细胞性贫血和地中海贫血。2)调查将 在体内进行蛋白质-DNA相互作用的珠蛋白 控制区让我们得以一窥珠蛋白基因的机制 体内控制，因为这些相互作用对组织特异性很重要 以及珠蛋白基因的发育特异性表达。反式- 与这些关键序列结合的作用蛋白将是 特色化的。这些研究将导致对控制的理解 珠蛋白基因表达和血红蛋白转换的关系，并可能提供 洞察新的治疗方式。3)交付方式 将珠蛋白基因导入红系细胞以实现高水平表达 将通过使用反式作用因子结合进行研究 增强珠蛋白基因表达的DNA基序。嵌合基因 在成人红细胞中持久表达丙种球蛋白也会 测试过。此外，腺相关病毒载体的使用将是 与逆转录病毒的传播相比。这三条途径 研究可能有助于预防纯合子疾病并导致新的 镰状细胞性贫血和地中海贫血的治疗体会。

DESCRIPTION: This proposal is a continuation of studies on normal and abnormal synthesis in hemoglobinopathies in sickle cell anemia and thalassemia. These hereditary anemias affect people from wide geographic areas, including the Mediterranean region, Africa,Middle East, Indian subcontinent and Southeast Asia. Studies will concentrate on three different areas which will contribute to the improved management of these disorders. 1) Research in the past grant years has indicated that the reverse dot blot is the most simple nonradioactive method for detecting mutations in sickle cell anemia. Hence, during coming years, analyses will be designed to cover most of the mutations in different ethnic groups and geographic areas in order to expedite prenatal diagnosis. Also, because fetal cells are known to cross into maternal circulation early in pregnancy, research will be performed on simple methods of isolating fetal cells from mother s blood in order to provide a noninvasive form of fetal DNA test for sickle cell anemia and thalassemia. 2) Investigations will be conducted on in vivo protein-DNA interactions between the globin control regions to allow a glimpse into the mechanism of globin gene control in vivo, as these interactions are important for tissue-specific and developmental-specific expression of the globin genes. The trans- acting proteins that are bound to these critical sequences will be characterized. These studies will lead to the understanding of control of globin gene expression and hemoglobin switching and may provide insight into new treatment modalities. 3) Methods of delivery of globin genes into erythroid cells to allow high level of expression will be investigated through the use of trans-acting factor binding DNA motifs which enhance globin gene expression. Chimeric genes which express gamma globin persistently in the adult red cell will also be tested. Also, the use of adeno-associated virus vectors will be compared to retroviral delivery. These three avenues of investigation may help to prevent homozygous disease and lead to new insight in the treatment of sickle cell anemia and thalassemia.