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

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

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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.

项目摘要囊性纤维化（CF）是由CFTR基因突变引起的，是最常见的遗传之一儿童疾病，影响美国出生的4,000名儿童中有1个（www.cff.org）。今天，CF疾病症状在具有最常见突变（∆F508 CFTR）的患者中，可以通过三种不同的药物改善组合。但是，药物具有负面影响，可以减少患者对治疗的依从性方案和构成长期健康风险。此外，许多其他常见的CF引起的突变做出了反应当前所有CF药物的所有CF患者仅与有症状的治疗。由于缺乏蛋白质结构将揭示这些变体的构象缺陷，主要是由于技术表达和净化足够数量的这些不稳定蛋白质的困难。表征不折叠的CFTR变体的构象缺陷并帮助开发新疗法，我们以前开发了共价蛋白绘画（CPP），这是一种用于体内结构表征的新方法质谱法蛋白质。在这里，我们建议开发一种更敏感和多重的CPP方法，命名为BioTMT-CPP，可以促进CFTR构象变化的检测和比较样品。新方法将有助于构象缺陷的表征错误折叠的CFTR 对当前疗法难治性的变体。此外，我们的方法有可能查明药物结合位点并确定当前CF药物的作用机理，这三个是未知的四个活性化合物。这种知识将有助于使药物组合疗法合理化。我们也建议在功能上表征了我们发现CPP的新型CFTR构象，在蛋白质运输过程中可能折叠率和无活性CFTR。深入了解分子机制稳定这种构象，以及将其释放到主动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。