GH Alternative Splicing: Mechanisms and Disease

GH 选择性剪接：机制和疾病

基本信息

批准号：
7418326
负责人：
John Atlas Phillips III
金额：
$ 38.49万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1984
资助国家：
美国
起止时间：
1984-08-01 至 2010-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7418326
关键词：
Affect Affinity Chromatography Alleles Alternative Splicing Anterior Pituitary Gland Blood Circulation Cell Death Cells Classification Count Cultured Cells Cystic Fibrosis Disease Dominant-Negative Mutation Enhancers Ensure Exons GH1 gene Genes Genetic Polymorphism Genomics Genotype Goals Haplotypes Hereditary Breast Carcinoma Hereditary Disease Human Individual Injection of therapeutic agent Lead Leukocytes Link Mediating Missense Mutation Molecular Chaperones Mus Mutation Nonsense-Mediated Decay Partner in relationship Patients Pattern Phenotype Pituitary Gland Point Mutation Protein Isoforms Proteins Proteome RNA RNA Interference RNA Splicing Regulatory Element Research Personnel Small Interfering RNA Somatotropin Source Testing Therapeutic Agents Transcript Transcriptional Silencer Elements Transgenic Organisms Variant Work enhancer binding protein gene therapy genetic regulatory protein growth hormone deficiency mouse model novel therapeutics prevent programs research study small molecule

项目摘要

DESCRIPTION (provided by applicant): Splicing is essential for expression of most genes and alternative splicing is a major source of human proteome diversity. At least 15% of all disease causing point mutations, not counting genomic variants that affect intronic or exonic splicing enhancer or silencer elements derange splicing. Many important diseases including cystic fibrosis and familial breast cancer have been linked to rare genomic variants (mutations) and common polymorphisms that alter splicing to produce abnormal, deleterious protein products. Understanding and correcting aberrant splicing caused by genomic variants could prevent expression of disease causing products and lead to new therapies for many genetic diseases. RNA interference with siRNAs can block expression of isoforms caused by numerous splicing mutations making it an attractive therapeutic agent for many genetic disorders, especially dominant negative diseases. Analysis of growth hormone genes from individuals with GH deficiency shows that aberrant skipping of exon 3 can produce a dominant negative 17.5kD isoform. In previous studies the P.I. found that at least three splicing enhancers are needed to ensure correct exon 3 definition and inclusion. The current work is now focused on determining the mechanisms of splicing enhancer function and how rare or common Growth Hormone 1 variants alter splicing to cause GHD or short stature. They will test new therapies to prevent GHD in mouse models where increased 17.5kD levels block secretion of wild type GH to cause cell death in pituitary somatotrophs. To achieve these goals the P.I, plans the following Specific Aims: 1) Determine how GH1 splicing enhancers activate exon 3 inclusion; 2) Determine if allele specific siRNAs can restore secretion in isolated Growth Hormone Deficiency II; 3) Develop new therapeutic approaches to treat Isolated Growth Hormone Deficiency II and 4) Determine how rare GH1 variants cause GHD and the contribution of common GH1 variations to GHD phenotype and short stature.

描述（由申请人提供）：剪接对于大多数基因的表达至关重要，而替代剪接是人类蛋白质组多样性的主要来源。在所有疾病中，至少有15％引起点突变，而不是计算影响内含子或外部剪接增强子或消音器元素剪接的基因组变异。包括囊性纤维化和家族性乳腺癌在内的许多重要疾病都与罕见的基因组变异体（突变）和常见的多态性有关，这些变体改变了剪接以产生异常，有害的蛋白质产物。理解和纠正由基因组变异引起的异常剪接可以防止表达引起产物的疾病并导致许多遗传疾病的新疗法。 RNA对siRNA的干扰可以阻止由许多剪接突变引起的同工型的表达，从而使其成为许多遗传疾病（尤其是显性阴性疾病）的有吸引力的治疗剂。对患有GH缺乏症个体的生长激素基因的分析表明，外显子3的异常跳过会产生显性负17.5KD同工型。在先前的研究中发现至少需要三个剪接增强器来确保正确的外显子3定义和包含。现在，目前的工作重点是确定剪接增强子函数的机制，以及稀有或常见的生长激素1变体如何改变剪接以引起GHD或短身材。他们将测试新的疗法，以防止小鼠模型中的GHD，其中增加了17.5kD水平的野生型GH的分泌，从而导致垂体营养营养的细胞死亡。为了实现这些目标，P.I，计划以下特定目的：1）确定GH1剪接增强器如何激活外显子3包含； 2）确定等位基因特异性siRNA是否可以恢复孤立生长激素缺乏症中的分泌II； 3）开发新的治疗方法来治疗孤立的生长激素缺乏率II和4）确定罕见的GH1变体如何引起GHD以及常见的GH1变异对GHD表型和矮小的身材的贡献。