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：
10478214
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/10478214
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 Epithelial Cells Exons FDA approved Future Gene Expression Genes Genetic Genotype Health Hereditary Disease Human Individual Interruption Intervention Investigation 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 Techniques Terminator Codon Testing Therapeutic Transcript Transfer RNA Transforming Growth Factor beta Translating Translations Untranslated RNA Untranslated Regions VX-770 airway epithelium airway surface liquid antagonist 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

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 Epithelial Cells Exons FDA approved Future Gene Expression Genes Genetic Genotype Health Hereditary Disease Human Individual Interruption Intervention Investigation 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 Techniques Terminator Codon Testing Therapeutic Transcript Transfer RNA Transforming Growth Factor beta Translating Translations Untranslated RNA Untranslated Regions VX-770 airway epithelium airway surface liquid antagonist 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

展开

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.

项目摘要囊性纤维化（CF），一种生命限制的常染色体隐性肺疾病，影响3500中的1个儿童是由囊性纤维化跨膜电导调节剂突变引起的（CFTR）基因。最近，FDA批准的CFTR调节器部分改善了该肺功能最普遍的突变（F508DEL），但其他常见的基因型，例如过早终止密码子（PTC）废话突变缺乏有效的干预。即使在F508DEL中患者对最新三重组合调制器鸡尾酒的响应者低25％。我们发现一个称为miR-145的小型非编码microRNA调节CFTR基因表达并阻止CFTR调制器的功效。我们以前的数据表明完成 miR-145拮抗作用改善了F508DEL治疗。该项目将采取更精确的策略利用反义寡核苷酸（ASO）在空间上仅阻碍miR-145与 CFTR而不会中断其他途径。我们假设ASO指导的miR-145目标位点阻断可改善CFTR校正。我们提出了3个目标：特定目标＃1：通过ASO封锁选择性增强F508DEL CFTR校正 miR-145结合位点的。特定目的＃2：通过MiR-145操纵在低处加强CFTR校正响应者。特定目的＃3：采用miRNA抑制来增强PTC中的通读功效突变。这些目的共同研究miR-145抑制作用，作为改善接下来的新策略生成CFTR校正。 AIM 1测试ASO目标站点封锁的功效和选择性为了增强F508DEL CF气道上皮细胞和相关动物的CFTR调节剂响应型号。 AIM 2使用来自低反应者到FDA批准的CFTR的患者来源的鼻样样本调节器测试miR-145抑制是否会提高个人利益。 AIM 3使用 miR-145抑制以增加可用于突变中PTC读取的CFTR底物目前缺乏有效的疗法。这些实验将检查出小说，高度选择性推进CFTR指导的治疗剂的策略，在CFTR干预中广泛适用和患者基因型。