Modulation of Jagged1/Pitpna in DMD as a means of therapy

DMD 中 Jagged1/Pitpna 的调节作为治疗手段

基本信息

批准号：
10666524
负责人：
LOUIS M KUNKEL
金额：
$ 38.94万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10666524
关键词：
Affect Bioinformatics Blood flow Canis familiaris Cardiopulmonary Cell Line Cell model Cessation of life Clinical Research Complement Control Animal DNA Sequence Alteration Data Disease Disease Progression Down-Regulation Duchenne muscular dystrophy Dystrophin Evaluation Exhibits Failure Fibrosis Gene Expression Profiling Generations Genes Genetic Genetic study Goals Grant Human In Vitro Injections Investigation Life Longevity Modeling Mus Muscle Muscle Cells Muscle function Muscular Atrophy Muscular Dystrophies Myoblasts Outcome PTEN gene Pathology Pathway interactions Patient-Focused Outcomes Patients Performance Phenotype Phosphatidylinositol Transfer Protein Predisposition Proliferating Proteins Proto-Oncogene Proteins c-akt Reading Frames Replacement Therapy Research Role Sarcolemma Signal Pathway Signal Transduction Skeletal Muscle Swimming Symptoms Terminator Codon Testing Therapeutic Transgenic Organisms Translations Viral Zebrafish adeno-associated viral vector candidate identification comparative disease-causing mutation efficacy evaluation exon skipping experimental study follow-up heart function improved in vivo inhibitor inhibitor therapy knock-down mRNA Expression mini-dystrophin mouse model muscle degeneration muscular structure overexpression pharmacologic premature prevent restoration targeted treatment therapeutic target treatment strategy

项目摘要

PROJECT SUMMARY/ABSTRACT Duchenne Muscular Dystrophy (DMD) is a degenerative muscle wasting disease caused by mutations in the dystrophin gene. The absence of dystrophin protein at the muscle sarcolemma results in increased muscle susceptibility to contraction-induced damage as well as dysregulation of secondary signaling pathways, many of which remain poorly understood. Despite advances in treatment strategies under investigation aimed at restoration of dystrophin expression including viral delivery of mini-dystrophin, read-through of translation stop codons, and exon skipping to restore the reading frame, there is no cure for DMD, and the identification of therapies that improve pathology independent of dystrophin would be of significant value to patients. In this vain, genetic modifiers of DMD are emerging as potential therapeutic targets. Our lab recently identified Jagged1 and Pitpna as genetic modifiers of DMD pathology in a Golden Retriever Muscular Dystrophy (GRMD) dog colony in which two exceptional “escaper” dogs exhibited a drastically milder phenotype than typical GRMD dogs despite being dystrophin-deficient. Normally, GRMD dogs show a severe phenotype similar to human DMD including early progressive muscle degeneration, fibrosis, and premature death often within the first 2 years of life due to cardiopulmonary failure. Gene expression analyses of the escaper dogs compared to severely affected GRMD dogs and control animals revealed that Jagged1 overexpression (OE) and decreased Pitpna expression were hallmarks of the mild phenotype, which including maintained ambulation and normal lifespan. In subsequent genetic studies of dystrophin-deficient zebrafish, we demonstrated that modulation of Jagged1 and Pitpna prevents manifestation of the dystrophic muscle phenotype, increases long-term survival, and improves swim performance. In primary myoblasts derived from normal and DMD patients, we also showed that Jagged1 overexpression and Pitpna inhibition impact AKT/PTEN signaling and improve myoblast fusion. Given this positive data, we will now extend our research of Jagged1 and Pitpna modulation into the well-characterized mdx5cv mouse model of DMD. We will also investigate genetic and pharmacological inhibition of PDE10A, which we have shown to elicit decreased Pitpna expression and improve dystrophic pathology in dystrophin-deficient zebrafish. The long-term goal in this project is to assess and validate the therapeutic potential of the genetic modifiers Jagged1 and Pitpna to ameliorate DMD pathology, and to identify a viable pharmacologic modulator to advance into clinical studies. To accomplish this, we will implement experiments in accordance to the following three specific aims: 1) Determine the effect of reduced Pitpna expression on dystrophic pathology in mdx5cv mice, 2) Characterize the functional role of Jagged1 overexpression in mdx5cv mice, and 3) Assess the therapeutic potential of PDE10A inhibition on dystrophic pathology in primary human muscle cells, zebrafish, and mouse models of DMD.

项目摘要/摘要 Duchenne肌肉营养不良（DMD）是由突变引起的变性肌肉浪费疾病肌营养不良基因。在肌肉肌膜上缺乏肌营养不良蛋白蛋白会导致肌肉增加对合同引起的损害以及次级信号通路失调的敏感性，许多其中的理解仍然很差。尽管在调查的治疗策略方面取得了进步恢复肌营养不良蛋白表达密码子和外显子跳过恢复阅读框，无法治愈DMD，识别与肌营养不良蛋白无关的改善病理学的疗法对患者具有重要的价值。在这个徒劳，DMD的遗传修饰符正在成为潜在的治疗靶标。我们的实验室最近确定锯齿状1和PITPNA作为黄金猎犬肌肉营养不良中DMD病理学的遗传修饰剂（GRMD）狗殖民地，其中两只特殊的“ Escaper”狗暴露了一个巨大的米勒表型典型的GRMD狗目的地是肌营养不良的。通常，GRMD狗显示出严重的表型类似于人类DMD，包括早期进行性肌肉变性，纤维化和过早死亡在生命的头两年内，由于心肺衰竭。 Escaper犬的基因表达分析与受到严重影响的GRMD犬和对照动物相比，发现锯齿状1过表达（OE） PITPNA表达的扩展是温和表型的标志，其中包括保持行动和正常寿命。在随后的肌营养不良蛋白斑马鱼的遗传研究中，我们证明了锯齿状1和pitpNA的调节可防止营养不良的肌肉表现表型，增加长期生存并改善游泳表现。在主要的成肌细胞中正常患者和DMD患者，我们还表明锯齿状1过表达和PITPNA抑制作用 AKT/PTEN信号传导并改善成肌细胞融合。鉴于这些积极数据，我们现在将扩展我们的研究 JAGGED1和PITPNA调制DMD的良好表征MDX5CV小鼠模型。我们也会研究PDE10A的遗传和药物抑制作用，我们已证明pITPNA下降了表达并改善了肌营养不良斑点斑马鱼中的营养不良病理学。长期目标项目是评估和验证遗传修饰符JAGGED1和PITPNA的治疗潜力改善DMD病理学，并确定可行的药物调节剂以进展到临床研究。为此，我们将根据以下三个特定目的实施实验：1）确定PITPNA表达降低对MDX5CV小鼠营养不良病理学的影响，2）表征 JAGGED1过表达在MDX5CV小鼠中的功能作用，3）评估PDE10A的治疗潜力 DMD的原发性肌肉细胞，斑马鱼和小鼠模型对营养不良病理的抑制作用。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Skeletal Muscle MicroRNAs: Their Diagnostic and Therapeutic Potential in Human Muscle Diseases.

DOI：
10.3233/jnd-140058
发表时间：
2015
期刊：
Journal of neuromuscular diseases
影响因子：
3.3
作者：
Alexander MS;Kunkel LM
通讯作者：
Kunkel LM

Tandem duplication within the DMD gene in Labrador retrievers with a mild clinical phenotype.

DOI：
10.1016/j.nmd.2022.08.001
发表时间：
2022-10
期刊：
NEUROMUSCULAR DISORDERS
影响因子：
2.8
作者：
Shelton, G. Diane;Minor, Katie M.;Vieira, Natassia M.;Kunkel, Louis M.;Friedenberg, Steven G.;Cullen, Jonah N.;Guo, Ling T.;Zatz, Mayana;Mickelson, James R.
通讯作者：
Mickelson, James R.

Pharmacological therapeutics targeting the secondary defects and downstream pathology of Duchenne muscular dystrophy.

DOI：
10.1080/21678707.2016.1240613
发表时间：
2016
期刊：
Expert opinion on orphan drugs
影响因子：
0.8
作者：
Spinazzola JM;Kunkel LM
通讯作者：
Kunkel LM

Diagnostic capabilities of nanopore long-read sequencing in muscular dystrophy.

DOI：
10.1002/acn3.51612
发表时间：
2022-08
期刊：
Annals of clinical and translational neurology
影响因子：
5.3
作者：
通讯作者：

Jagged 1 Rescues the Duchenne Muscular Dystrophy Phenotype.

DOI：
10.1016/j.cell.2015.10.049
发表时间：
2015-11-19
期刊：
Cell
影响因子：
64.5
作者：
Vieira NM;Elvers I;Alexander MS;Moreira YB;Eran A;Gomes JP;Marshall JL;Karlsson EK;Verjovski-Almeida S;Lindblad-Toh K;Kunkel LM;Zatz M
通讯作者：
Zatz M