Antisense oligonucleotide treatment for Pompe disease

庞贝病的反义寡核苷酸治疗

基本信息

批准号：
10433785
负责人：
VIRGINIA Eunice KIMONIS
金额：
$ 20.72万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
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：
2022-07-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10433785
关键词：
Address Age Age-Months Antisense Oligonucleotides Applications Grants Autophagocytosis Base Pairing Cardiomyopathies Clinical Trials Combined Modality Therapy Control Groups Cytoplasm DNA Sequence Data Dose Drug Delivery Systems Early treatment Enzymes Evaluation Fabry Disease Fibrosis Future Gaucher Disease Genes Genetic Diseases Glucan 1,4-alpha-Glucosidase Glycogen Glycogen (Starch) Synthase Glycogen Storage Disease Glycogen storage disease type II Goals Government Heart Histologic Histology Human Impairment In Vitro Knock-in Knock-out Lead Lysosomal Storage Diseases Lysosomes Measurement Measures Mediating Messenger RNA Mus Muscle Muscle Weakness Muscle function Myocardium Myopathy Outcome Pathologic Patients Persons Pharmacologic Substance Production RNA Recombinants Residual state Respiratory Diaphragm Single-Stranded DNA Skeletal Muscle Spinal Muscular Atrophy Technology Testing Therapeutic Tissue Stains Tissues Treatment Efficacy Wild Type Mouse Work base clinical application design drug efficacy effective therapy efficacy study enzyme activity enzyme replacement therapy gene replacement glucosidase improved innovation knock-down mouse model muscle degeneration muscle strength novel preclinical study prevent response small molecule standard of care success translational study treatment group treatment strategy

项目摘要

Project Abstract: Pompe disease is a devastating myopathy resulting from the deficiency of the lysosomal enzyme acid alpha- glucosidase (GAA),resulting in accumulation of glycogen in lysosomes and cytoplasm. The standard of care is enzyme replacement therapy (ERT) with recombinant human (rh) GAA. Patients with Pompe disease often have poor outcomes due to limited efficacy of the drug in clearing muscle glycogen stores. Substrate Reduction Therapy (SRT) is effective in other lysosomal storage diseases such as Gaucher and Fabry disease. We propose that by knocking down glycogen synthase (GSY1), we will reduce glycogen synthesis in muscle, thus reducing the amount of substrate requiring to be cleared by enzyme. Our long-term goal is to develop a potent therapy to stop/reduce the progression of the muscle disease in Pompe disease. Major gaps: Despite ERT, patients continue to accumulate glycogen in lysosomes leading to muscle fibrosis and fatty replacement. Effective therapy for the progressive muscle weakness of Pompe disease is thus a huge unmet need. Preliminary results: Antisense Oligonucleotides (ASO) technology has emerged as a powerful direct therapeutic alternative to conventional small molecule approaches or gene replacement strategies for the treatment of genetic disorders such as spinal muscular atrophy. ASOs are short, synthetic single-stranded DNA sequences designed to target RNA by well-characterized Watson-Crick base pairing, and once bound to the target RNA, can modulate RNA function through a degradative mechanism using RnaseH1. Ionis Pharmaceuticals designed and screened ASOs targeting mouse GYS1 in vitro to identify the most efficacious ASO. The lead ASOs from the screen were validated in a dose response study and the top 10 ASOs were screened for testing in wild type mice. Three GYS1 ASOs showed the best tolerability and efficacy profile leading to knock down of GYS1 mRNA by approximately 50% of control. Preclinical studies of the effects of the GYS1 ASOs 25 mg/kg/weekly on muscle glycogen, histology, and muscle function in 1 month and 3-month-old Pompe mice indicated that GYS1 ASOs were effective in knocking down GYS1 by approximately 85%, with approximately 50% improvement in clearing muscle glycogen levels. We therefore believe that early reduction of glycogen substrate or a combination therapy with ERT may be the key to successful treatment in Pompe disease. Hypothesis: We propose that GYS1 ASOs in Pompe mice will reduce glycogen production and prevent muscle weakness, either as monotherapy if started early, or in combination with ERT, to obtain maximum therapeutic benefit. Specific Aims: Aim 1: Identification of the most effective ASO-mediated glycogen synthase (GYS1) knock down to prevent glycogen accumulation in a mouse model of Pompe disease. Aim 2: Correction of muscle glycogen and weakness using GYS1 antisense oligonucleotides in combination with Enzyme Replacement Therapy (ERT) in Pompe mice. Success with this translational study will pave the way for novel effective treatments for Pompe disease.

项目摘要：庞贝疾病是由于溶酶体酶α-缺乏而导致的造成灾难性肌病。葡萄糖酶（GAA），导致糖原在溶酶体和细胞质中积累。护理标准是重组人（RH）GAA的酶替代疗法（ERT）。庞贝疾病的患者经常患有由于药物在清除肌肉糖原储存方面的疗效而导致的结果不佳。底物还原治疗（SRT）在其他溶酶体储存疾病（如高彻和法布里病）中有效。我们建议通过击倒糖原合酶（GSY1），我们将减少肌肉中的糖原合成，从而减少需要用酶清除的底物量。我们的长期目标是开发有效的疗法停止/减少庞贝疾病中肌肉疾病的进展。主要差距：尽管ERT，患者继续在溶酶体中积聚糖原，导致肌肉纤维化和脂肪替代。有效的疗法因此，庞贝疾病的进行性肌肉无力是巨大的未满足需求。初步结果：反义寡核苷酸（ASO）技术已成为强大的直接治疗替代方案用于治疗遗传疾病的常规小分子方法或基因替代策略例如脊柱肌肉萎缩。 ASOS是简短的合成单链DNA序列，设计为靶向通过特征良好的Watson-Crick底座配对RNA，一旦与目标RNA结合，可以调节RNA 通过使用RNAseH1的降解机制功能。 Ionis Pharmaceuticals设计和筛选了ASOS 在体外靶向小鼠Gys1以识别最有效的ASO。屏幕上的线索是在一项剂量反应研究中验证和前10个ASO在野生型小鼠中进行了测试。三 Gys1 ASOS显示出最佳的耐受性和功效概况，从而导致击倒Gys1 mRNA 大约50％的对照。 GYS1 ASOS 25 mg/kg/每周对肌肉的影响的临床前研究糖原，组织学和肌肉功能在1个月和3个月大的庞贝小鼠中表明GYS1 ASOS 有效地将GYS1击倒约85％，清算提高了约50％肌肉糖原水平。因此，我们认为糖原底物或联合疗法的早期还原使用ERT可能是成功治疗庞贝疾病的关键。假设：我们提出Gys1 ASOS 在蓬松中，小鼠将减少糖原的产生并防止肌肉无力，如果开始早期或与ERT结合使用，以获得最大的治疗益处。特定目的：目标1：身份证明最有效的ASO介导的糖原合酶（GYS1）敲低以防止糖原积累在庞贝疾病的小鼠模型中。 AIM 2：使用GYS1纠正肌肉糖原和无力庞贝小鼠中的反义寡核苷酸与酶替代疗法（ERT）结合使用。成功通过这项翻译研究，将为庞贝疾病的新有效治疗铺平道路。