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: 8828567
• 负责人: LOUIS M KUNKEL
• 金额: $ 38.83万
• 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/8828567
• 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; Health; 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; research study; skeletal; transcriptome sequencing; wasting

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：Duchenne肌肉营养不良（DMD）是由肌营养不良蛋白基因突变引起的一种退化性肌肉浪费疾病。肌肉中缺乏肌营养不良蛋白的蛋白会导致次级信号通路失调，其中许多人的理解水平仍然很少。 Mayana Zatz博士的实验室已经发现了两只肌营养不良蛋白缺陷的狗，这些狗来自金毛犬肌肉营养不良（GRMD）狗，这些狗受到了轻度影响，并逃脱了早期的致死性和营养不良表型（肌肉），这些表型（肌肉浪费）正常发生在受影响的GRMD犬中。我们对这些狗和其他GRMD和对照犬的微阵列分析显示，PITPNA基因的大幅减少，PitpNA基因是PTEN/AKT信号通路的已知调节剂。在使用DMD患者进行的人类活检的MicroRNA微阵列的并行研究中，我们确定了MicroRNA MiR-486，该mir-486在DMD肌肉中仅降低，并已显示出我们和其他人可以改变PTEN/AKT信号。我们假设miR-486的过表达和/或PITPNA抑制会导致PTEN水平降低，同时随后增加磷酸化（激活的）AKT，这可能导致疾病病程的严重程度降低。预计诱导磷酸化-AKT（由miR-486和PITPNA调节引起）会影响该途径的几个下游靶基因，这将导致肌肉肥大，血管结构基因血管性基因的增加，肌肉结构基因，肌肉肌肉连接点的增加，抑制了APOPTSIS的神经连接点，并有效基因，并有效基因（APOPTRETIC GENERS/APCTRETS PENTREMENS），并且有效基因（Aptrecter pten）（pten pten）（pten）的基因（有效）（pten Strem pten）。如utrophin）。我们计划实现我们的长期目标，即通过以下三个具体目的调节MiR-486和/或PITPNA（MDX5CV）模型中MiR-486和/或PITPNA的表达来改变DMD中的信号传导途径：1）表征miR-486和PITPNA在Vitro中PTEN/AKT信号通路的调节。 2）miR-486表达的操纵以调节肌营养不良蛋白缺陷肌肉中的PTEN/AKT信号传导，以建模未来的治疗剂。 3）在肌肉中pitpna水平降低的小鼠以及其对PTEN/AKT信号的影响的表征。