MANIPULATION OF PTEN/AKT SIGNALING IN DUCHENNE MUSCULAR DYSTROPHY AS A MEANS OF T

杜氏肌营养不良症中 PTEN/AKT 信号传导的调控作为 T 的一种手段

• 批准号: 8631323
• 负责人: LOUIS M KUNKEL
• 金额: $ 38.61万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8631323
• 关键词: AKT Signaling Pathway; Affect; Animal Model; Biopsy; Canis familiaris; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cell Survival; Cells; Clinical; Coupled; Data; Disease; Disease Progression; Doxycycline; Duchenne muscular dystrophy; Dystrophin; Future; Gene Targeting; Generations; Genes; Goals; Growth; Human; In Vitro; Inhibition of Apoptosis; Injection of therapeutic agent; Knockout Mice; Laboratories; Lead; Link; MicroRNAs; Microarray Analysis; Modeling; Mus; Muscle; Muscle Cells; Muscular Dystrophies; Mutation; Myopathy; Neuromuscular Junction; Outcome; PTEN gene; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Physiology; Production; Proteins; Proto-Oncogene Proteins c-akt; Publishing; Regulation; Role; Route; Severity of illness; Signal Pathway; Signal Transduction; Skeletal Muscle; Structural Genes; Subfamily lentivirinae; Symptoms; Tamoxifen; Testing; Therapeutic; Therapeutic Uses; Transgenic Mice; Utrophin; Vascularization; Wallerian Degeneration; Wasting Syndrome; Zebrafish; adeno-associated viral vector; boys; cell growth; combinatorial; comparative; disease-causing mutation; dosage; functional improvement; improved; in vivo; mRNA Expression; mouse model; muscle degeneration; muscle hypertrophy; novel; novel therapeutic intervention; overexpression; public health relevance; research study; skeletal; transcriptome sequencing; wasting

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 in muscle results in dysregulated secondary signaling pathways, many of which remain poorly understood. The laboratory of Dr. Mayana Zatz has identified two dystrophin-deficient dogs from a litter of Golden Retriever Muscular Dystrophy (GRMD) dogs that are mildly affected and escape the early lethality and dystrophic phenotype (muscle wasting) that normally occur in severely affected GRMD dogs. Our microarray analysis of these dogs and other GRMD and control dogs, revealed a strong reduction of the Pitpna gene, which is a known regulator of the PTEN/AKT signaling pathway. In parallel studies using microRNA microarrays performed on human biopsies from patients with DMD, we identified a microRNA, miR-486, that is exclusively reduced in DMD muscle and has been shown by us and others to alter PTEN/AKT signaling. We hypothesize that either overexpression of miR-486 and/or inhibition of Pitpna results in decreased PTEN levels while subsequently increasing phosphorylated (activated) AKT, which might lead to reduced severity of disease course. The induction of phosphorylated-AKT (resulting from miR-486 and Pitpna modulation), are predicted to affect several downstream target genes of this pathway which will result in an increase in muscle hypertrophy, muscle structural genes vascularization genes, the increase in the neuromuscular junction number of branch points, inhibition of apoptosis genes, and validated PTEN/AKT downstream target muscle genes (such as Utrophin). We plan to approach our long-term goal of understanding how these two alter signaling pathways in DMD by modulating expression of miR-486 and/or Pitpna in the mouse (mdx5cv) model according to the following three specific aims: 1) Characterization of miR- 486 and Pitpna regulation of PTEN/AKT signaling pathway in vitro. 2) Manipulation of miR-486 expression to modulate PTEN/AKT signaling in dystrophin-deficient muscle for modeling of future therapeutics. 3) Generation of mice with reduced Pitpna levels in muscle and characterization of its effects on PTEN/AKT signaling.

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。肌肉中缺乏肌营养不良蛋白蛋白会导致次级信号失调 途径，其中许多途径仍然很少理解。 Mayana Zatz博士的实验室已经确定了两个 肌营养不良的狗来自金毛犬肌肉营养不良症（GRMD）狗的狗 受影响并逃脱通常发生在 严重影响的GRMD狗。我们对这些狗和其他GRMD和对照狗的微阵列分析， 揭示了PITPNA基因的强烈降低，该基因是PTEN/AKT信号的已知调节剂 路径。在使用MicroRNA微阵列的并行研究中，对患者的人类活检进行了 DMD，我们确定了一个microRNA，miR-486，该mir-486在DMD肌肉中仅降低，并已通过 我们和其他人会改变PTEN/AKT信号。我们假设mir-486和/或 抑制PITPNA导致PTEN水平降低，同时增加磷酸化（激活） AKT，这可能导致疾病病程的严重程度降低。诱导磷酸化蛋白（由此产生 从miR-486和PITPNA调制中），预计会影响该途径的几个下游靶基因 这将导致肌肉肥大的增加，肌肉结构基因血管化基因，即 分支点的神经肌肉连接数增加，抑制细胞凋亡基因并进行了验证 PTEN/AKT下游目标肌肉基因（例如Utrophin）。我们计划实现我们的长期目标 了解这两个通过调节miR-486和/或的表达来改变DMD中的信号通路 根据以下三个特定目的，小鼠（MDX5CV）模型中的PITPNA：1）mir-表征 PTEN/AKT信号通路的486和PITPNA调节体外。 2）miR-486表达对 在肌营养不良蛋白缺陷肌肉中调节PTEN/AKT信号传导，以建模未来的治疗剂。 3） 在肌肉中的pitpna水平降低的小鼠以及其对PTEN/AKT的影响的表征 信号。