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）：杜氏肌营养不良症（DMD）是一种由肌营养不良蛋白基因突变引起的退行性肌肉萎缩疾病。肌营养不良蛋白在肌肉中的缺乏导致调节失调的次级信号通路，其中许多仍然知之甚少。Mayana Zatz博士的实验室已经从一窝金毛寻回犬肌营养不良症（GRMD）犬中鉴定出两只肌营养不良蛋白缺陷犬，这些犬受到轻度影响，并逃脱了通常发生在严重受影响的GRMD犬中的早期致死性和营养不良表型（肌肉萎缩）。我们对这些狗和其他GRMD和对照狗的微阵列分析揭示了Pitpna基因的强烈减少，Pitpna基因是PTEN/AKT信号通路的已知调节因子。在使用microRNA微阵列对DMD患者的人类活检进行的平行研究中，我们鉴定了一种microRNA，miR-486，它只在DMD肌肉中减少，并且已经被我们和其他人证明可以改变PTEN/AKT信号传导。我们假设miR-486的过表达和/或Pitpna的抑制导致PTEN水平降低，同时随后增加磷酸化（活化）AKT，这可能导致疾病进程的严重程度降低。预测磷酸化AKT的诱导（由miR-486和Pitpna调节产生）影响该途径的几个下游靶基因，这将导致肌肉肥大、肌肉结构基因血管化基因、分支点的神经肌肉接头数量增加、凋亡基因的抑制和验证的PTEN/AKT下游靶肌肉基因（如Utrophin）。我们计划根据以下三个具体目标来实现我们的长期目标，即理解这两者如何通过调节小鼠（mdx 5cv）模型中的miR-486和/或Pitpna的表达来改变DMD中的信号通路：1）体外表征miR- 486和Pitpna对PTEN/AKT信号通路的调节。2)操纵miR-486表达以调节肌营养不良蛋白缺陷肌肉中的PTEN/AKT信号传导，用于未来治疗的建模。3)肌肉中Pitpna水平降低的小鼠的产生及其对PTEN/AKT信号传导的影响的表征。