The CFTR Interactome

CFTR 相互作用组

基本信息

批准号：
10677830
负责人：
John R Yates III
金额：
$ 54.01万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-05 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10677830
关键词：
Affect Age Anions Applications Grants Attenuated Biogenesis Cells Cessation of life Child Childhood Clinical Trials Code Combination Drug Therapy Combined Modality Therapy Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator DNA Insertion Elements Defect Detection Development Disease Drug Binding Site Drug Combinations Drug Targeting Effectiveness Epithelium Exclusion FDA approved Goals Health Homeostasis Immunofluorescence Immunologic Inherited Knowledge Label Left Light Lung Mass Spectrum Analysis Methods Molecular Molecular Conformation Mutation Names Nature Outcome Paint Patients Peptides Pharmaceutical Preparations Pharmacotherapy Phosphorylation Point Mutation Post-Translational Protein Processing Protein Conformation Proteins Rationalization Refractory Regimen Regulator Genes Risk Sampling Science Signal Transduction Site Sodium Chloride Solvents Stains Structural defect Structure Symptoms Therapeutic Therapeutic Intervention Tissues VX-809 Variant Western Blotting Work carbene compliance behavior cystic fibrosis patients design drug development improved in vivo innovation insight mass spectrometer mutant novel novel therapeutic intervention novel therapeutics oxidation protein structure protein transport side effect technology development three dimensional structure trafficking

项目摘要

Project Summary Cystic Fibrosis (CF) is caused by mutation of the CFTR gene and is one of the most common inherited childhood diseases, impacting 1 in 4,000 children born in the US (www.cff.org). Today, CF disease symptoms in patients with the most common mutation (∆F508 CFTR) can be improved with three different drug combinations. However, the drugs have negative side effects that reduce patient compliance to therapeutic regimens and pose long-term health risks. Additionally, many other common CF-causing mutations respond poorly or not at all to any of the current CF drugs, leaving CF patients carrying these mutations only with symptomatic therapy. Efforts to develop new compounds for such CF variants are hampered by the lack of protein structures that would reveal the conformational defects of these variants, mostly due to technical difficulties in expressing and purifying sufficient quantities of these unstable proteins. To characterize the conformational defects of misfolded CFTR variants and to aid in the development of new therapies, we previously developed Covalent Protein Painting (CPP), a novel method for in vivo structural characterization of proteins by mass spectrometry. Here, we propose to develop a more sensitive and multiplexable CPP method, named bioTMT-CPP, that will facilitate detection and comparison of CFTR conformational changes between samples. The new method will facilitate the characterization of conformational defects in misfolded CFTR variants that are refractory to current therapies. Furthermore, our approach has the potential to pinpoint drug binding sites and identify the mechanism of action of current CF drugs, which remain unknown for three of the four active compounds. Such knowledge will help to rationalize drug combination therapies. We also propose to functionally characterize a novel CFTR conformation that we discovered by CPP and that is attained by misfolded and inactive CFTR, likely during protein trafficking. Insight into the molecular mechanisms that stabilize this conformation as well as those that release it into an active conformation will be invaluable for further corrector drug development and will benefit all CF patients.

项目概要囊性纤维化 (CF) 是由 CFTR 基因突变引起的，是最常见的遗传性疾病之一如今，CF 疾病症状影响着美国出生的每 4,000 名儿童中的 1 名 (www.cff.org)。具有最常见突变（ΔF508 CFTR）的患者可以通过三种不同的药物得到改善然而，这些药物具有副作用，会降低患者对治疗的依从性。此外，许多其他常见的导致 CF 的突变也会产生反应。目前任何 CF 药物的效果都很差或根本没有效果，使得携带这些突变的 CF 患者只能接受由于缺乏药物，开发针对此类 CF 变体的新化合物的努力受到阻碍。蛋白质结构将揭示这些变体的构象缺陷，主要是由于技术原因表达和纯化足够数量的这些不稳定蛋白质的困难。错误折叠的 CFTR 变体的构象缺陷并帮助开发新疗法，我们先前开发的共价蛋白质绘画（CPP），一种体内结构表征的新方法在这里，我们建议开发一种更灵敏且可多重使用的 CPP 方法，命名为bioTMT-CPP，这将有助于检测和比较之间的CFTR构象变化新方法将有助于错误折叠 CFTR 中构象缺陷的表征。此外，我们的方法有可能精确定位药物。结合位点并确定当前 CF 药物的作用机制，其中三种药物尚不清楚我们还提出了四种活性化合物的知识将有助于合理化药物组合疗法。对我们通过 CPP 发现并通过以下方法获得的新型 CFTR 构象进行功能表征错误折叠和失活的 CFTR，可能是在蛋白质运输过程中发生的。稳定该构象以及将其释放为活性构象的构象对于进一步的校正药物开发将使所有 CF 患者受益。