Ribosomal perturbation as a mechanism to prevent misfolding of CFTR

核糖体扰动作为防止 CFTR 错误折叠的机制

基本信息

  • 批准号:
    10063541
  • 负责人:
  • 金额:
    $ 55.62万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2017
  • 资助国家:
    美国
  • 起止时间:
    2017-12-15 至 2022-01-07
  • 项目状态:
    已结题

项目摘要

Abstract Cystic Fibrosis (CF) is a common lethal autosomal recessive disorder, occurring in 1 per 2,500 to 3,500 U.S. births annually. The disease is caused by defects in the CF transmembrane conductance regulator (CFTR). Mutations that change the CFTR coding sequence alter integrity of peptide folding and lead to disease. In this project, we show that CFTR codon alterations—including both non-synonymous and synonymous polymorphisms on background of the common F508del variant—can perturb ribosome dynamics, consequent mRNA utilization, translational rate, and protein biogenesis. The critical relationship between protein folding and translational velocity is a topical area with ramifications ranging from basic CFTR trafficking to disease phenotype and intervention. Mechanistic underpinnings for this application are derived from genome-wide phenomic screening with the Saccharomyces cerevisiae deletion strain library to identify novel modulators of F508del CFTR maturation, leading to discovery of specific ribosomal protein modules that impact F508del CFTR folding. We have established that suppression of Rpl12, along with other 60S proteins comprising the ribosomal stalk, improve F508del CFTR processing and activity at the mammalian cell surface to levels (in primary airway epithelia) predicted to confer clinical benefit among CF patients and comparable in magnitude to lumacaftor, a new agent recently approved for this purpose. Our preliminary data indicate that improved folding and activity of F508del CFTR are attributable to effects on translational kinetics. We propose three Specific Aims to develop an innovative model relevant to CF pathogenesis: 1) Define ribosomal domains and functional pathway interactions that govern ∆F protein biogenesis, and expand the analysis to include other CFTR variants and complex alleles, 2) Ascertain the mechanism by which Rpl12 suppression rescues F508del CFTR function, including relevance of translation rate to disease causing CF mutations and other CFTR polymorphisms, 3) Determine in vivo significance by development of RPL12 conditional knockout or haplosufficient mice. Our team combines multidisciplinary and mutually reinforcing expertise in CFTR biochemistry, transport physiology, ribosome profiling, translational velocity, yeast phenomics, and molecular genetics to mechanistically address a fundamental hypothesis regarding ways the ribosome utilizes mRNA to influence folding and functional quality of resulting gene products. We will establish translation control as a novel and critical mediator of Yor1 and CFTR maturation, identify specific ribosome modules that influence the CFTR biogenesis pathway, and evaluate relevance and safety of repressing this target in a murine disease model. Such results will improve understanding of CF molecular mechanism, suggest novel approaches for personalized treatment of CF patients with the most common forms of the disease, and indicate clinical significance of an important new observation relevant to aberrant protein translation and a fatal genetic illness.
抽象的 囊性纤维化(CF)是一种常见的致死常染色体隐性疾病,发生在美国2500至3,500美国。 每年出生。该疾病是由CF跨膜电导调节剂(CFTR)缺陷引起的。 改变CFTR编码序列的突变改变了胡椒折叠的完整性并导致疾病。在这个 项目,我们表明CFTR密码子更改 - 包括非同义词和同义词 公共F508DEL变体背景上的多态性 - 可以扰动核糖体动力学,因此 mRNA利用,翻译速率和蛋白质生物发生。蛋白质折叠之间的关键关系 翻译速度是一个局部区域,分析范围从基本CFTR运输到疾病 表型和干预。该应用的机械基础来自全基因组 用酿酒酵母缺失菌株库进行现象筛查,以识别新型调节剂 F508DEL CFTR成熟,导致发现影响F508DEL的特定核糖体蛋白模块 CFTR折叠。我们已经确定了RPL12的抑制作用,以及其他60年代蛋白质完成 核糖体茎,改善哺乳动物细胞表面的F508DEL CFTR加工和活性至水平(在 原发性气道上皮)预计将赋予CF患者的临床益处,并且幅度可比 致Lumachaftor,最近为此目的批准了新的代理商。我们的初步数据表明改进了 F508DEL CFTR的折叠和活性归因于对翻译动力学的影响。我们提出了三个 特定目的是开发与CF发病机理相关的创新模型:1)定义核糖体域和 控制∆F蛋白生物发生的功能途径相互作用,并扩展分析以包括其他 CFTR变体和复杂等位基因,2)确定RPL12抑制响应F508DEL的机制 CFTR功能,包括转化率与引起CF突变和其他CFTR的疾病的相关性 多态性,3)通过开发RPL12有条件敲除或 单倍体的小鼠。我们的团队结合了CFTR中的多学科和相互加强的专业知识 生物化学,运输生理学,核糖体分析,翻译速度,酵母菌学和分子 关于核糖体利用mRNA的方式,机械地解决基本假设的遗传学 影响所得基因产物的折叠和功能质量。我们将建立翻译控制 YOR1和CFTR成熟的新颖和关键介体,确定影响影响的特定核糖体模块 CFTR生物发生途径,并评估反映鼠类疾病中该靶标的相关性和安全性 模型。这样的结果将改善对CF分子机制的理解,提出了新的方法 对具有最常见疾病形式的CF患者的个性化治疗,并表明临床 与异常蛋白质翻译和致命遗传疾病有关的重要新观察的重要性。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

暂无数据

数据更新时间:2024-06-01

JOHN L HARTMAN的其他基金

Discovery of novel mechanisms that impact CFTR translation and contribute to cystic fibrosis pathogenesis
发现影响 CFTR 翻译并导致囊性纤维化发病机制的新机制
  • 批准号:
    10367064
    10367064
  • 财政年份:
    2017
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Discovery of novel mechanisms that impact CFTR translation and contribute to cystic fibrosis pathogenesis
发现影响 CFTR 翻译并导致囊性纤维化发病机制的新机制
  • 批准号:
    10545091
    10545091
  • 财政年份:
    2017
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Core B - Research Development Core
核心 B - 研究开发核心
  • 批准号:
    10633282
    10633282
  • 财政年份:
    2015
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Constructing gene-regulatory networks to reveal the metabolic basis of lifespan i
构建基因调控网络揭示寿命的代谢基础
  • 批准号:
    8372173
    8372173
  • 财政年份:
    2012
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Constructing gene-regulatory networks in yeast for a metabolic basis of lifespan
在酵母中构建基因调控网络作为寿命的代谢基础
  • 批准号:
    8535594
    8535594
  • 财政年份:
    2012
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Constructing gene-regulatory networks to reveal the metabolic basis of lifespan in yeast
构建基因调控网络以揭示酵母寿命的代谢基础
  • 批准号:
    9099632
    9099632
  • 财政年份:
    2012
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Constructing gene-regulatory networks to reveal the metabolic basis of lifespan in yeast
构建基因调控网络以揭示酵母寿命的代谢基础
  • 批准号:
    8871509
    8871509
  • 财政年份:
    2012
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Constructing gene-regulatory networks to reveal the metabolic basis of lifespan i
构建基因调控网络揭示寿命的代谢基础
  • 批准号:
    8721828
    8721828
  • 财政年份:
    2012
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Molecular Buffering of Replication in Yeast
酵母复制的分子缓冲
  • 批准号:
    6689967
    6689967
  • 财政年份:
    2001
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:
Molecular Buffering of Replication in Yeast
酵母复制的分子缓冲
  • 批准号:
    7023873
    7023873
  • 财政年份:
    2001
  • 资助金额:
    $ 55.62万
    $ 55.62万
  • 项目类别:

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