An investigation into the CFTR-Associated Ligand (CAL) and the underlying molecular mechanism by which it increases cell surface expression of ΔF508-CFTR through regulation of trafficking pathways

对 CFTR 相关配体 (CAL) 及其通过调节运输途径增加 αF508-CFTR 细胞表面表达的潜在分子机制的研究

基本信息

批准号：
8912065
负责人：
Emily Anne Bergbower
金额：
$ 0.73万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-07 至 2015-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) cause Cystic Fibrosis, a common lethal autosomal recessive disorder, by affecting the water/salt balance of the lungs and several other key organs. The CFTR-Associated Ligand (CAL) is a PDZ domain binding protein that binds to the C-terminus of CFTR. While it has been previously shown that CAL reduces cell surface WT-CFTR and promotes its degradation in the lysosome, CAL's interactions with ΔF508-CFTR have never been characterized. The goal of this study is to illuminate how CAL regulates trafficking and degradation of ΔF508-CFTR. The present proposal posits that CAL is critical for ER and post-ER trafficking of the ΔF508-CFTR mutant and may be a worthwhile target for treatment in patients. The proposal is built upon two specific aims, the first of which is to elucidate to the physiological role of CAL in the early trafficking pathway of ΔF508-CFTR. The second aim is focused on determining which accessory proteins, including chaperones and well known CAL interacting proteins, are influenced by or bound to CAL in the process of ΔF508-CFTR regulation. Preliminary data has shown that CAL increases cell surface expression of ΔF508-CFTR and is degraded in the proteasome. The research training program has been designed for successful completion of the stated aims in two years, as the student has completed all other necessary degree program requirements and already has promising preliminary data. Methods for achieving the stated goals of the research are focused on cell culture work and microscopy, both of which are easily accessible within the Johns Hopkins School of Medicine and thus are realistic in scope.

描述（由申请人提供）：囊性纤维化跨膜电导调节器（CFTR）的突变通过影响肺部和其他几个关键器官的水/盐平衡而导致囊性纤维化，这是一种常见的致死性常染色体隐性遗传病。 (CAL) 是一种 PDZ 结构域结合蛋白，可与 CFTR 的 C 末端结合，此前已表明该蛋白。虽然 CAL 会减少细胞表面 WT-CFTR 并促进其在溶酶体中的降解，但 CAL 与 ΔF508-CFTR 的相互作用尚未得到表征。本研究的目的是阐明 CAL 如何调节 ΔF508-CFTR 的运输和降解。 CAL 对于 ΔF508-CFTR 突变体的 ER 和 ER 后转运至关重要，并且可能是患者治疗的一个有价值的目标该提案建立在两个特定的基础上。第一个目标是阐明 CAL 在 ΔF508-CFTR 早期运输途径中的生理作用，第二个目标是确定哪些辅助蛋白（包括分子伴侣和众所周知的 CAL 相互作用蛋白）受到影响或结合。初步数据显示，CAL 增加了 ΔF508-CFTR 的细胞表面表达，并在蛋白酶体中被降解。研究培训计划旨在在两年内成功完成既定目标，因为学生已完成所有其他必要的学位课程要求，并且已经拥有实现既定研究目标的有希望的初步数据，重点是细胞培养工作。和显微镜，这两种技术在约翰·霍普金斯大学医学院内都很容易获得，因此在范围上是现实的。