Comprehensive mapping of trafficking and functional robustness in Inward Rectifier K+ channels for variant pathogenicity prediction and model-guided engineering of chemogenetic reagents

全面绘制内向整流器 K 通道中的运输和功能鲁棒性，用于化学遗传学试剂的变异致病性预测和模型引导工程

• 批准号: 10297049
• 负责人: Daniel Schmidt
• 金额: $ 31.66万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10297049
• 关键词: Amino Acids; Benchmarking; Benign; Biological Assay; Cardiovascular Diseases; Cell Line; Cell surface; Cells; Clinical; Coupled; Data; Data Set; Defect; Diabetes Mellitus; Disease; Engineering; Etiology; Flow Cytometry; GIRK1 subunit, G protein-coupled inwardly-rectifying potassium channel; Genotype; Ion Channel; Ions; Libraries; Ligands; Link; Machine Learning; Maps; Membrane Potentials; Metabolic Diseases; Missense Mutation; Modeling; Molecular; Molecular Chaperones; Mutagenesis; Mutation; Neurons; Pathogenicity; Peptides; Pharmaceutical Preparations; Phenotype; Play; Potassium Channel; Property; Proteins; Reagent; Regulation; Resources; Rest; Role; Sirolimus; Structure; Structure-Activity Relationship; Surface; Tertiary Protein Structure; Tissues; Trimethoprim; Variant; addiction; base; biophysical model; data acquisition; human disease; inward rectifier potassium channel; nervous system disorder; neuromuscular; next generation sequencing; novel therapeutics; operation; predictive modeling; programs; technological innovation; tool; trafficking; treatment strategy

Project Summary Inward Rectifier K+ channels (KIR) play key roles in the operation of cells in neuromuscular and other tissue. Pathogenic variants are linked to numerous neurological, cardiovascular, and metabolic disorders. Although some variants cause gating defects in KIR by altering ligand regulation or ion permeation, there is growing evidence that many –perhaps most– variants cause defects in folding and trafficking of KIR. Despite the central role for folding and trafficking in the disease etiology, there have been to date no comprehensive large-scale studies that determine sequence and structural determinants of KIR trafficking and functional robustness. Here we provide the first comprehensive assessment of missense and topological mutations’ effects on KIR trafficking and function. Acquisition of these data is the required first step to build quantitative biophysical model of the sequence, structure, and function relationship in KIR. These models will be useful to understand the mechanistic basis for KIR mutation phenotypes, to predict their pathogenicity, and to identify new treatment strategies for KIR- linked disorders. These models will also pave the way for rational engineering of KIR as chemogenetic reagents that can be used to study functional roles of K+ channels in intact tissues.

项目摘要 内向整流器K+通道（KIR）在神经肌肉和其他组织中细胞的操作中起关键作用。 致病性变异与许多神经系统，心血管和代谢性疾病有关。虽然 某些变体通过改变配体调节或离子渗透会导致KIR的门控缺陷，并有增长 证据表明，许多变体可能会导致KIR折叠和贩运的缺陷。尽管有中央 折叠和贩运在疾病病因中的作用，迄今为止还没有全面的大规模 确定KIR运输和功能鲁棒性的序列和结构决定剂的研究。这里 我们对错义和拓扑突变对KIR贩运的影响提供了首次全面评估 和功能。获取这些数据是建立定量生物物理模型的第一步 KIR中的序列，结构和功能关系。这些模型将对理解机制有用 KIR突变表型的基础，预测其致病性，并确定KIR- 链接的疾病。这些模型还将为KIR作为化学试剂的合理工程铺平道路 可以用来研究K+通道在完整组织中的功能作用。