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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，是最常见的遗传性疾病之一。儿童疾病，影响美国出生的4,000名儿童中的1名（www.cff.org）。今天，CF疾病的症状在最常见的突变患者（CFTR 508）中，可以用三种不同的药物改善组合。然而，这些药物具有负面的副作用，降低了患者对治疗的依从性。并造成长期的健康风险。此外，许多其他常见的CF引起的突变，对目前的任何CF药物都很差或根本不起作用，使携带这些突变的CF患者仅具有对症治疗开发用于此类CF变体的新化合物的努力受到缺乏蛋白质结构，将揭示这些变体的构象缺陷，主要是由于技术表达和纯化足够量的这些不稳定蛋白质的困难。表征错误折叠的CFTR变体的构象缺陷，并帮助开发新的治疗方法，我们先前开发的共价蛋白绘画（CPP），一种用于体内结构表征的新方法，蛋白质质谱在这里，我们建议开发一种更灵敏和可复用的CPP方法，命名为bioTMT-CPP，这将有助于检测和比较CFTR之间的构象变化，样品新方法将有助于表征错误折叠的CFTR中的构象缺陷对当前疗法难治的变体。此外，我们的方法有可能确定药物结合位点，并确定目前CF药物的作用机制，这仍然是未知的三个四种活性化合物这些知识将有助于合理化药物联合治疗。我们亦建议功能上表征了一种新的CFTR构象，我们发现了CPP，并获得了错误折叠和失活的CFTR，可能在蛋白质运输过程中。深入了解分子机制稳定该构象以及将其释放成活性构象那些构象对于进一步的校正药物开发，将有利于所有CF患者。