MOLECULAR BASIS OF THE CYSTIC FIBROSIS PHENOTYPE

这项SCOR应用的目标是了解囊性纤维化(CF)中有缺陷的基因CFTR突变的临床、生物学和生化后果。该计划汇集了一批具有广泛专业知识的基础科学家和临床研究人员，通过研究患者的CFTR分子缺陷、生成小鼠模型、绘制修饰基因图谱以及使用细胞培养和体外蛋白质系统，从不同的角度分析CF疾病。我们结合了我们在CF患者记录、人类和小鼠遗传学、生物化学和细胞生物学领域的优势。SCOR分为4个研究项目(RP)、3个试点项目(PP)和3个核心单元，分为3个研究领域，即患者的临床和遗传学研究、识别CF修饰基因的小鼠模型和CFTR的直接表征。在第一个领域，RP1将建立对由疾病的主要和次要遗传决定因素引起的或与之相关的CF疾病表型谱的全面了解。在第二个实验中，RP2将研究ClC-2氯通道在小鼠模型中调节上皮氯分泌的作用，而RP3将在特定遗传背景下剖析CF小鼠肺部疾病的生理和遗传方面。此外，还包括一个名为PP3的试点项目，以表征最近在一个同源的CF小鼠品系中观察到的肝病。这些研究将发现新的途径，通过这些途径可以设计出治疗CF的替代方法。在第三个领域，Rp4将在分子、细胞和功能水平上进行详细的分析，以确定在CFTR的第一个核苷酸结合域(NBD1)上发生的错义突变的后果，以及导致羧基末端截断的突变将被用作这些研究的探针。这将得到两个试点项目的补充：PP1将探索一种新的荧光转移技术来研究跨膜片段相互作用，PP2将研究纯化的CFTR是否在重组体系中中介大的有机阴离子，如谷氨酸和谷胱甘肽的能量依赖的运输。除了管理核心，患者/生物统计核心和鼠标核心将为霍夫项目提供支持。SCORE的结果将对CF病理的分子机制产生新的见解，并将导致新的改进的治疗方法。

The goal of this SCOR application is to understand the clinical, biological and biochemical consequences of mutations in CFTR, the gene defective in cystic fibrosis (CF). The program brings together a group of basic scientists and clinician researchers with a broad range of expertise to the common task of analyzing the CF disease from different angles, through studying the CFTR molecular defects in patients, generating, mouse models, mapping of modifier genes, and using cell culture and in vitro systems for the protein. We combine our strengths in the area of CF patient documentation, human and mouse genetics, biochemistry and cell biology. The SCOR is organized into 4 Research Projects (RP), 3 Pilot Projects (PP) and 3 Core Units, grouped into 3 research areas, namely, clinical and genetic studies of patients, mouse models of identification of CF modifier genes, and direct characterization of CFTR. In the first area, RP1 will establish a comprehensive understanding of the spectrum of CF disease phenotype caused by or associated with the primary and secondary genetic determinants of the disease. In the second, RP2 will study the role of ClC-2 chloride channels in mediating epithelial chloride secretion in a mouse model and RP3 will dissect the physiologic and genetic aspects of lung disease in CF mice of a specific genetic background. In addition, a pilot project, PP3, is included to characterize the liver disease recently observed in one of the congenic CF mouse strains. These studies will discover new pathways through which alternative methods may be devised to treat CF. In the third area, RP4 will pursue a detailed analysis at the molecular, cellular and functional levels to establish the consequences of the missense mutations that occur in the first nucleotide binding domain (NBD1) of CFTR and mutations causing carboxyl terminal truncations will be used as probes for these studies. This will be complemented by the two pilot projects: PP1 which will explore a new fluorescence transfer technique for the study of transmembrane segment interactions and PP2 which will examine if purified CFTR an mediate energy-dependent transport of large organic anions such as glutamate and glutathione in a reconstituted system. In addition to the Administration Core, the Patient/Biostatistics Core, and the Mouse Core will serve to support the hove projects. The results from Score should yield novel insights into the molecular mechanisms of CF pathology and should lead to new improved therapeutic approaches.