Oligotherapeutics to enhance CFTR correction

寡疗法增强 CFTR 校正

基本信息

批准号：
10000198
负责人：
William Thomas Harris
金额：
$ 7.43万
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：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10000198
关键词：
3&apos Untranslated Regions Address Affect Alleles Animals Antisense Oligonucleotides Area Award Binding Binding Sites Biochemical Biological Assay Bronchoalveolar Lavage CCL2 gene Cellularity Child Childhood Clinic Collaborations Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Delta F508 mutation Development Dose Epithelial Epithelial Cells Epithelium FDA approved Foundations Fruit Genetic Genotype Growth Human Immune Tolerance Immunohistochemistry In Vitro Inbred CFTR Mice Interleukin-1 beta Interleukin-6 Interruption Intervention Irrigation Laboratories Length Liquid substance Lung Lung diseases Measures Mediating Messenger RNA MicroRNAs Modeling Mutation Oligonucleotides Parents Pathway interactions Pharmaceutical Preparations Pharmacologic Substance Pharmacology Protein Glycosylation Proteins Pulmonary Fibrosis Pulmonary Function Test/Forced Expiratory Volume 1 Reporter Research Respiratory physiology Rodent Safety Seeds Severity of illness Site Specificity Spinal Muscular Atrophy Structure of respiratory epithelium System TGF Beta Signaling Pathway TNF gene Technology Testing Therapeutic Transcript Transforming Growth Factor beta Translations Treatment Efficacy Untranslated Regions Vertebral column airway epithelium career cystic fibrosis patients cytokine drug development experimental study improved in vitro Model in vivo in vivo Model in vivo evaluation innovation mutant neutrophil novel novel strategies phosphorothioate preclinical efficacy preservation protein expression response stereochemistry therapeutic development therapeutic target tool treatment response

项目摘要

PROJECT SUMMARY Cystic Fibrosis (CF) is a fatal lung disease that affects 1 in 3500 children. F508del CFTR, the most common mutation present in 90% of CF patients, has proven difficult to treat. The two FDA approved drugs, Orkambi® (lumacaftor and ivacaftor) and Symdeko® (tezacaftor and ivacaftor), only improve lung function by 2-4%. Improving F508del CFTR remains a critical unmet need in CF therapeutics. One barrier to F508del CFTR correction is increased expression of the genetic modifier TGF-β in CF lung and airway epithelia. TGF-β suppresses CFTR function and nullifies the benefit of Orkambi® and Symdeko®. Our laboratory has discovered that the microRNA miR-145 mediates TGF-β inhibition of CFTR correction. TGF-β increases miR-145 which directly binds to the 3'-untranslated region (3'-UTR) to degrade CFTR transcripts and diminish protein expression. Loss of CFTR substrate eliminates therapeutic response. miR-145 antagonism overcomes this barrier to improve Orkambi® benefit. The project pursues a novel approach to augment F508del therapeutics. We will utilize an antisense oligonucleotide (ASO) to block miR-145 binding to CFTR. We hypothesize that ASO- directed miR-145 target site blockade improves F508del CFTR correction. Specific Aim #1: To selectively augment F508del CFTR correction through antisense oligonucleotide blockade of the miR-145 binding site. Specific Aim #2: To test in vivo delivery, safety, and efficacy of antisense oligonucleotide miRNA target specific blockade in humanized CFTR mice. These Aims will provide the necessary “next steps” in oligotherapeutic development. Aim 1 provides the in vitro mechanism, dosing, and efficacy data to establish the rationale for oligonucleotide targeting of the miRNA binding site to improve F508del CFTR correction. Aim 2 utilizes the recently developed full-length humanized CFTR mouse to establish delivery feasibility, safety profile, and therapeutic response in vivo.

项目摘要囊性纤维化（CF）是一种致命的肺部疾病，影响3500名儿童中有1例。 F508DEL CFTR，证明90％的CF患者中最常见的突变很难治疗。两个 FDA批准的药物，Orkambi®（Lumacaftor和Ivacaftor）和Symdeko®（Tezacaftor和Symdeko® ivacaftor），仅将肺功能提高2-4％。改善F508DEL CFTR仍然是关键 CF疗法未满足的需求。 F508DEL CFTR校正的一个障碍是遗传修饰符的表达增加 CF肺和气道上皮中的TGF-β。 TGF-β抑制CFTR功能并无效 Orkambi®和Symdeko®的好处。我们的实验室发现MicroRNA mir-145 介导TGF-β抑制CFTR校正。 TGF-β增加了直接结合的miR-145 3'-非翻译区域（3'-UTR）降低CFTR成绩单并减少蛋白质表达。 CFTR底物的丢失消除了热反应。 mir-145拮抗作用克服了这一障碍，以提高Orkambi®福利。该项目采用新颖的方法来增强F508DEL疗法。我们将利用一个反义寡核苷酸（ASO）阻断与CFTR结合的miR-145。我们假设这是定向miR-145目标部位封锁可改善F508DEL CFTR校正。特定目标＃1：通过反义选择性增强F508DEL CFTR校正 miR-145结合位点的寡核苷酸阻滞。特定目的＃2：测试反义寡核苷酸的体内递送，安全性和效率人源化CFTR小鼠中的miRNA靶标特异性阻滞。这些目标将为寡疗法开发提供必要的“下一步”。目标1 提供体外机制，剂量和效率数据，以建立理由 miRNA结合位点的寡核苷酸靶向改善F508DEL CFTR校正。目标2 利用最近开发的全长人源化CFTR鼠标建立输送在体内可行性，安全性和治疗反应。