Therapeutic Modulation of Myotonic Muscular Dystrophy

强直性肌营养不良症的治疗调节

基本信息

批准号：
10222788
负责人：
CHARLES A THORNTON
金额：
$ 37.06万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10222788
关键词：
3&apos Untranslated Regions Ablation Affect Aftercare Age Alleles Alternative Splicing Antisense Oligonucleotides Binding Proteins Biochemical Biogenesis Biological Biological Assay CAG repeat CUG repeat Cardiac Cell Culture Techniques Cell Extracts Cell Nucleus Clinical Trials Coupled Cytoplasm DNA DNA Repair Defect Development Discrimination Disease Dose Exhibits Exonuclease Gene Frequency Gene Silencing Genes Genetic Transcription Goals Growth Human Hypersensitivity Individual Industry Injections Investigational Therapies Laboratories Learning Macaca fascicularis Mediating Modeling Mus Muscle Muscle Cells Muscle Fibers Muscle Weakness Muscular Dystrophies Mutation Myocardium Myopathy Myotonia Myotonic Muscular Dystrophy Myotonic dystrophy type 1 Nuclear Oligonucleotides Output Pathology Pharmaceutical Preparations Plant Roots Process Property Proteins RNA RNA Splicing Reagent Refractory Regimen Residual state Ribonuclease H Safety Single-Stranded DNA Site Skeletal Muscle Somatic Cell Subcutaneous Injections Symptoms Testing Therapeutic Therapeutic Trials Time Tissue Sample Tissues Toxic effect Transcript Transgenic Mice Work age related base clinical practice design gain of function in vivo Model knock-down mRNA Precursor mouse model mutant myotonic dystrophy protein kinase novel postnatal preservation prevent programs public health relevance systemic toxicity therapeutic candidate therapeutic development

项目摘要

DESCRIPTION (provided by applicant): Myotonic dystrophy type 1 (DM1) results from a novel RNA-mediated disease process that is triggered by an unconventional mutation. The mutation is an unstable expansion of CTG repeats in the DM protein kinase (DMPK) gene, and the mechanism involves a toxic gain-of-function by transcripts containing an expanded CUG repeat. Studies indicate that symptom onset and progression of DM1 is driven by the age-dependent growth of the expanded CTG repeat in somatic cells. In an effort to attack DM1 at its root cause, we have partnered with colleagues in industry to develop antisense oligonucleotides (ASOs) targeting the toxic RNA. This approach was highly effective in mouse models and has recently entered clinical trials. In this proposal we seek to better define the therapeutic properties of these versatile new reagents. First we will determine whether ASOs may exhibit allelic selectively; that is, whether they preferentially target transcripts containing expanded CUG repeats (CUGexp). Our previous work suggested that CUGexp-containing transcripts are hypersensitive to ASO knockdown, owing to prolonged dwell time in the nucleus. We developed a new mouse model for testing the allelic discrimination of ASOs, based on the presence or absence of expanded CUG repeats in the target transcript. The sensitivity of expanded vs. non-expanded alleles to ASO/RNase H knockdown will be compared, and the therapeutic window for allelic selectivity will be determined for different dosing regimens. The results will guide th development of therapeutic strategies to maximize and preserve the residual function of the DM kinase. Next we will determine whether somatic instability of expanded repeats is reduced by DMPK-targeting ASOs. Previously we showed that CAG-repeat ASOs are able to stabilize CTG repeats in a DM1 model in vivo. However, as the preferred therapeutic strategy has now shifted to targeting outside of the repeat tract, we need to determine whether ASOs directed at the non-repetitive sequences of DMPK may also stabilize expanded repeats. If ASOs do in fact exhibit dual properties of mitigating RNA toxicity and stabilizing repeats, this will argue for early initiation of treatment. Next, we will examine the biochemical basis for the unusually prolonged duration of ASO activity in muscle cells. Using decoy oligonucleotides that are complementary to ASOs, coupled with quantitative assays of pre-mRNA, we will determine whether long-duration activity results from post-transcriptional silencing and depends on the continuous presence and action of the ASO. Finally, we will determine whether early initiation of ASO treatment can prevent dystrophic pathology, spliceopathy, and muscle weakness in a CUGexp-expressing, MBNL1-deficient model. Taken together, the results of the proposed studies will expand our understanding of the biological properties of ASOs in skeletal muscle, and provide important guidance for the optimal use of these reagents in therapeutic trials and clinical practice.

描述（由适用提供）：肌发育症1型（DM1）是由新型RNA介导的疾病过程引起的，该过程是由非常规突变触发的。该突变是DM蛋白激酶（DMPK）基因中CTG重复序列的不稳定膨胀，该机制涉及通过包含含有扩展CUG cug重复膨胀的转录物获得的功能获得的功能。研究表明，DM1的症状发作和进展是由体细胞中CTG重复的年龄依赖性生长驱动的。为了攻击DM1的根本原因，我们与行业的同事合作开发了针对有毒RNA的反义寡核苷酸（ASOS）。这种方法在小鼠模型中非常有效，并且最近进入了临床试验。在此提案中，我们试图更好地定义这些多功能新试剂的治疗特性。首先，我们将确定ASO是否可以选择性地表现出等位基因；也就是说，他们是否更喜欢靶向包含扩展的CUG重复序列（CugeXP）的转录本。我们以前的工作表明，含CugeXP的转录本对ASO敲低是过敏的，这是由于核中的停留时间延长。我们开发了一种新的小鼠模型，以根据目标转录本中存在或不存在扩展的CUG重复序列来测试ASO的等位基因歧视。将比较对ASO/RNase H敲低的扩展和非扩展等位基因的敏感性，并将确定不同剂量方案的等位基因选择性的治疗窗口。结果将指导理论策略的发展，以最大化和保留DM激酶的残留功能。接下来，我们将通过靶向DMPK的ASO来确定是否会减少扩展重复的体细胞不稳定性。以前，我们表明CAG重复ASO能够在体内DM1模型中稳定CTG重复序列。但是，由于现在首选的理论策略已转移到重复道之外的靶向，因此我们需要确定针对DMPK非重复序列的ASO是否也可以稳定扩展的重复序列。如果ASOS实际上确实暴露了降低RNA毒性和稳定重复序列的双重性能，则这将主张尽早开始治疗。接下来，我们将检查肌肉细胞中异常活性持续时间异常延长的生化基础。使用与ASOS互补的诱饵寡核苷酸，再加上对MRNA的定量评估，我们将确定长期活性是否是由转录后沉默引起的，并取决于ASO的持续存在和作用。最后，我们将确定在表达CugeXP的MBNL1缺陷模型中，早期开始ASO治疗是否可以防止营养不良的病理，剪接和肌肉无力。综上所述，拟议的研究的结果将扩展我们对骨骼肌中ASO的生物学特性的理解，并为这些试剂在治疗试验和临床实践中的最佳使用提供了重要的指导。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Design of Bivalent Nucleic Acid Ligands for Recognition of RNA-Repeated Expansion Associated with Huntington's Disease.

用于识别与亨廷顿病相关的 RNA 重复扩增的二价核酸配体的设计。

DOI：
10.1021/acs.biochem.8b00062
发表时间：
2018
期刊：
Biochemistry
影响因子：
2.9
作者：
Thadke,ShivajiA;Perera,JDinithiR;Hridya,VM;Bhatt,Kirti;Shaikh,AshifY;Hsieh,Wei-Che;Chen,Mengshen;Gayathri,Chakicherla;Gil,RobertoR;Rule,GordonS;Mukherjee,Arnab;Thornton,CharlesA;Ly,DanithH
通讯作者：
Ly,DanithH

Targeting DMPK with Antisense Oligonucleotide Improves Muscle Strength in Myotonic Dystrophy Type 1 Mice.

DOI：
10.1016/j.omtn.2017.05.007
发表时间：
2017-06-16
期刊：
Molecular therapy. Nucleic acids
影响因子：
0
作者：
Jauvin D;Chrétien J;Pandey SK;Martineau L;Revillod L;Bassez G;Lachon A;MacLeod AR;Gourdon G;Wheeler TM;Thornton CA;Bennett CF;Puymirat J
通讯作者：
Puymirat J