REGULATION OF EXPRESSION OF ACID BETA-GLUCOSIDASE

酸性 β-葡萄糖苷酶表达的调节

• 批准号: 3237737
• 负责人: FRANCES SMITH
• 金额: $ 11.84万
• 依托单位: EUNICE KENNEDY SHRIVER CTR MTL RETARDATN
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-04-01 至 1996-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3237737
• 关键词: Gaucher's disease; beta glucosidases; biological models; developmental neurobiology; embryo /fetus; enzyme activity; gene expression; human subject; infant animal; laboratory mouse; leukocytes; messenger RNA; protein metabolism; transcription factor

A deficiency in acid beta-glucosidase (betaGlc) causes Gaucher disease, which ranges in severity from a neurologically devastating disease in infancy to a mild disease in adulthood. A thorough understanding of the mechanisms that control betaGlc expression is necessary to design future therapeutic interventions, particularly gene therapy. this proposal aims to elucidate the regulation of betaGlc expression, placing particular emphasis on neural cells and cells of the monocyte/macrophage lineage, which are adversely affected in Gaucher disease. Specifically, we will determine: (1) What mechanisms control basal levels of betaGlc activity in different tissues? We have shown already that betaGlc mRNA expression plays a major role in differential betaGlc expression. We now plan to determine the role of protein turnover in regulating betaGlc activity, and to elucidate the role of transcription and mRNA stability in controlling betaGlc mRNA levels. We also plan to identify the sequences that control betaGlc mRNA levels, and the proteins that bind these elements. Preliminary results indicate that the betaGlc promoter contains elements that are recognized by tissue specific and ubiquitous factors. (2) Does feedback regulation control betaGlc gene expression? BetaGlc activity in vivo increases in response to procedures that increase substrate level. We will establish an in vitro model in which the role of substrate accumulation on betaGlc expression can be evaluated. (3) How is betaGlc activity regulated in the developing nervous system? BetaGlc activity is high in mouse fetal brain and declines in the neonate. Using the mouse as a model system, betaGlc activity and mRNA levels will be measured during brain development. We will investigate how changes in the amounts of the transcription factors identified in aim #1 may influence betaGlc expression in vivo in the brain. Also, because it is possible that the developmental regulation of betaGlc may be driven by substrate levels, changes in the factors that control feedback regulation in neural cells elucidated in aim #2 also will be investigated. A detailed understanding of the control of betaGlc expression in neural cells has important implications not only for gene therapy of severe Gaucher disease, but also for the treatment of other neurological diseases that require gene expression in multiple cell types.

酸性β-葡萄糖苷酶(β-Glc)缺乏会导致高谢病， 其严重程度从神经学上的破坏性疾病到 从婴儿期到成年后的轻微疾病。透彻了解 控制BetaGlc表达的机制对于设计未来是必要的 治疗干预，特别是基因治疗。这项提议旨在 为了阐明BetaGlc表达的调节，将特定的 重点是神经细胞和单核/巨噬细胞系的细胞， 在高谢病中会受到不利影响。具体来说，我们将 确定：(1)什么机制控制基础水平的BetaGlc活性 在不同的组织里？我们已经证明了BetaGlc mRNA的表达 在不同的BetaGlc表达中起主要作用。我们现在计划 确定蛋白质周转在调节β-Glc活性中的作用， 并阐明转录和信使核糖核酸的稳定性在 控制BetaGlc基因水平。我们还计划确定这些序列 控制BetaGlc信使核糖核酸水平，以及结合这些基因的蛋白质 元素。初步结果表明，BetaGlc启动子 包含组织特定且无处不在的元素 各种因素。(2)反馈调控是否控制BetaGlc基因的表达？ 体内BetaGlc活性增加是对以下程序的响应 提高底物水平。我们将建立一个体外模型，其中 底物积累对BetaGlc表达的作用可以是 已评估。(3)BetaGlc活性在发育过程中是如何调节的 神经系统？小鼠胎脑中的BetaGlc活性很高 在新生儿中下降。使用鼠标作为模型系统，BetaGlc 活动和信使核糖核酸水平将在大脑发育期间进行测量。我们 将研究转录因子的数量是如何变化的 在AIM#1中发现的可能影响在体内的BetaGlc的表达 大脑。此外，因为发育调节有可能 可能是由底物水平驱动的，这些因素的变化 在Aim#2中还阐明了神经细胞中的控制反馈调节 将会被调查。详细了解BetaGlc的控制 神经细胞的表达不仅对基因有重要意义 治疗严重的高谢病，也用于治疗其他 需要在多个细胞中表达基因的神经系统疾病 类型。