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miR-145 target site blockade is a selective strategy to enhance CFTR restoration and readthrough

miR-145靶位点封锁是增强CFTR恢复和通读的选择性策略

基本信息

批准号：
10675757
负责人：
William Thomas Harris
金额：
$ 37.13万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10675757
关键词：
3&apos Untranslated Regions Affect Animal Model Antisense Oligonucleotides Base Sequence Binding Binding Sites CRISPR/Cas technology Caring Caucasians Cell Line Cells Child Chlorides Clinical Clinical Research Complex Cystic Fibrosis Data Defect Delta F508 mutation Disease Disease Progression Effectiveness Epithelial Cells Epithelium Exons FDA approved Future Gene Expression Genes Genetic Genotype Health Hereditary Disease Human Individual Interruption Intervention Length Life Lung diseases Measurement Measures Mediating Messenger RNA MicroRNAs Modeling Mucociliary Clearance Mucous body substance Mutation Nonsense Mutation Nonsense-Mediated Decay Nose Nucleotides Optical Coherence Tomography Organoids Participant Pathway Analysis Pathway interactions Patients Pharmaceutical Preparations Phase Phenotype Population Proteins Quality of life Rattus Regulator Genes Sampling Site Specimen Swelling Techniques Terminator Codon Testing Therapeutic Transcript Transfer RNA Transforming Growth Factor beta Translating Translations Untranslated RNA VX-770 airway epithelium airway surface liquid antagonist autosome cystic fibrosis airway epithelia cystic fibrosis patients disease-causing mutation effective intervention effective therapy efficacy testing experience experimental study falls genotyped patients improved in silico in vivo in vivo evaluation inhibitor innovation lung health monolayer mutant next generation novel novel strategies premature preservation protein expression pulmonary function response restoration screening side effect transcriptome sequencing

项目摘要

PROJECT SUMMARY Cystic fibrosis (CF), a life-limiting autosomal recessive lung disease that affects 1 in 3500 children, is caused by mutations in the Cystic Fibrosis Transmembane conductance Regulator (CFTR) gene. Recently FDA-approved CFTR modulators partially improve lung function for the most prevalent mutation (F508del), but other common genotypes, such as premature termination codon (PTC) nonsense mutations, lack an effective intervention. Even in F508del patients, 25% are low responders to the latest triple combination modulator cocktail. We discovered that a small non-coding microRNA called miR-145 regulates CFTR gene expression and blocks efficacy of CFTR modulators. Our previous data show that complete miR-145 antagonism improves F508del therapy. This project will pursue a more precise strategy that utilizes an antisense oligonucleotide (ASO) to sterically impede only miR-145 binding to CFTR without interrupting other pathways. We hypothesize that ASO-directed miR-145 target site blockade improves CFTR correction. We propose 3 Aims: Specific Aim #1: Enhance F508del CFTR correction selectively through ASO blockade of the miR-145 binding site. Specific Aim #2: Bolster CFTR correction through miR-145 manipulation in low responders. Specific Aim #3: Employ miRNA inhibition to augment readthrough efficacy in PTC mutations. Together, these Aims investigate miR-145 inhibition as a novel strategy to improve next- generation CFTR correction. Aim 1 tests the efficacy and selectivity of ASO target site blockade to enhance CFTR modulator response in F508del CF airway epithelial cells and relevant animal models. Aim 2 uses patient-derived nasal samples from low responders to FDA-approved CFTR modulators to test whether miR-145 inhibition boosts individual benefit. Aim 3 investigates using miR-145 inhibition to increase the CFTR substrate available for PTC readthrough in mutations that currently lack an effective therapy. These experiments will examine a novel, highly selective strategy to advance CFTR-directed therapeutics, broadly applicable across CFTR intervention and patient genotype.

项目概要囊性纤维化 (CF) 是一种限制生命的常染色体隐性肺病，影响每 3500 人中就有 1 人儿童，是由囊性纤维化跨膜电导调节器突变引起的（CFTR）基因。最近 FDA 批准的 CFTR 调节剂可部分改善肺功能最常见的突变 (F508del)，但其他常见基因型，例如早产儿终止密码子（PTC）无义突变，缺乏有效的干预措施。即使在 F508del 中患者中，25% 对最新的三重组合调节剂鸡尾酒反应较低。我们发现一种名为 miR-145 的小型非编码 microRNA 调节 CFTR 基因表达并阻断 CFTR 调节剂的功效。我们之前的数据显示完整 miR-145 拮抗作用可改善 F508del 治疗。该项目将追求更精准的策略利用反义寡核苷酸 (ASO) 仅在空间上阻碍 miR-145 与 CFTR 不中断其他途径。我们假设 ASO 指导的 miR-145 靶位点封锁改善了 CFTR 校正。我们提出 3 个目标：具体目标#1：通过 ASO 封锁有选择地增强 F508del CFTR 校正 miR-145 结合位点。具体目标#2：通过低水平的 miR-145 操作加强 CFTR 校正响应者。具体目标 #3：利用 miRNA 抑制来增强 PTC 的通读功效突变。这些目标共同研究 miR-145 抑制作为改善下一步的新策略：生成 CFTR 校正。目标 1 测试 ASO 靶点封锁的功效和选择性增强 F508del CF 气道上皮细胞和相关动物中 CFTR 调节剂的反应模型。目标 2 使用来自对 FDA 批准的 CFTR 低反应者的患者鼻腔样本调节剂来测试 miR-145 抑制是否会提高个体效益。目标 3 调查使用抑制 miR-145 可增加可用于突变中 PTC 通读的 CFTR 底物目前缺乏有效的治疗方法。这些实验将检验一种新颖的、高度选择性的推进 CFTR 导向治疗的策略，广泛适用于 CFTR 干预和患者基因型。