Generation of Wunen/LPP3-based therapy for muscular dystrophy

基于 Wunen/LPP3 的肌营养不良疗法的产生

• 批准号: 7941020
• 负责人: Hannele RUOHOLA-BAKER
• 金额: $ 46.76万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7941020
• 关键词: Address; Adult; Affect; Animal Disease Models; Apoptosis; Apoptotic; Area; Basal lamina; Binding; Biological Models; Canis familiaris; Cell Maintenance; Cell Proliferation; Cell Therapy; Cell Transplants; Cells; Collaborations; Complex; Defect; Discipline; Disease; Drosophila genus; Drosophila melanogaster; Dystrophin; Fibroblasts; Funding; Future; Gene Dosage; Generations; Genetic; Genetic Screening; Glycoproteins; Goals; Grant; Homologous Gene; Human; Knowledge; Laboratories; Link; Lipids; Metabolism; Methods; Modeling; Mus; Muscle; Muscle Cells; Muscle Fibers; Muscle function; Muscle satellite cell; Muscular Dystrophies; Musculoskeletal; Myoblasts; Natural regeneration; Pathway interactions; Patients; Phenotype; Phospholipids; Phosphoric Monoester Hydrolases; Positioning Attribute; Regenerative Medicine; Regulation; Role; Sarcolemma; Second Messenger Systems; Signal Transduction; Skeletal Myoblasts; Skin Tissue; Sphingolipids; Stem cells; System; Testing; Therapeutic; United States National Institutes of Health; Wasting Syndrome; Work; base; disease phenotype; fly; human embryonic stem cell; improved; in vivo; in vivo Model; induced pluripotent stem cell; lipid phosphate phosphatase; mdx mouse; mouse model; muscle degeneration; muscle regeneration; mutant; regenerative; research study; second messenger; sphingosine 1-phosphate; tissue regeneration

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (11) Regenerative Medicine and specific Challenge Topic 11-AR-101: Musculoskeletal and Skin Tissue Regeneration. The focus of this grant is to develop new strategies for muscular dystrophy therapies or/and strengthen the existing strategies. This grant is based on a new finding showing that reduction of LPP3 that reduces the level of active second messenger, S1P can suppress dystrophic phenotypes in Drosophila melanogaster. We will now study the mechanism of sphingolipid pathway and Dystrophin interactions and further, will test for wunen/LPP3 based strategies to ameliorate muscle function in dystrophic mice. Muscular Dystrophy is a muscle wasting disease that at present has no cure. As mentioned, our studies have produced a possible new inroad into treatment for this disease and it involves wunen. We will exhaustively characterize wunen, a suppressor isolated in our previous forward genetic screens for modifiers of the Dystrophin-associated Glycoprotein Complex mutant phenotypes. Wunen is a homolog of human This grant is based on a new finding showing that reduction of LPP3 that reduces the level of active second messenger, S1P can suppress dystrophic phenotypes in Drosophila melanogaster. We will now study the mechanism of sphingolipid pathway and Dystrophin interactions and further, will test for wunen/LPP3 based strategies to ameliorate muscle function in dystrophic mice. Muscular Dystrophy is a muscle wasting disease that at present has no cure. As mentioned, our studies have produced a possible new inroad into treatment for this disease and it involves wunen (LPP3) which many studies have shown is involved in the regulation of levels of bioactive lipids, particularly sphingosine 1-phosphate (S1P). S1P is implicated in pre-myoblast (adult muscle stem cell) maintenance as well as myoblast differentiation into myotubes. The possibility of a muscle regenerative strategy for suppressing the muscular dystrophy phenotype through the alteration of sphingolipid signaling has high potential for being extremely fruitful. We will determine if wunen acts through S1P to inhibit apoptosis and/or increase muscle generation through either increasing proliferation of adult muscle precursor cells or increasing proliferation of adult muscle satellite stem cells which have yet to be defined in Drosophila. We will determine if muscle satellite stem cells exist in the same physical niche as mammalian muscle satellite stem cells which is between the basal lamina and the sarcolemma of existing muscle fibers. In toto, we will establish and extend the role of sphingolipid signaling in muscle formation in Drosophila and determine the mechanism that alleviates the disease phenotype of dystrophic flies. Furthermore, we will test whether reduction of Wunen/LPP3 can be advantageous in muscular dystrophy to mammalian animal models of the disease, particularly the mdx mouse (and in the future the dog model). We hope to produce regenerative therapeutic strategies to alleviate the disease phenotype in this model. These strategies will include reducing LPP3 function as well as increasing the presence of its downstream effector S1P. We propose that increasing sphingolipid signaling will increase the regeneration potential of muscle stem cells and myoblasts and thereby will improve the efficacy of muscular dystrophy therapy. We will test this hypothesis by two different approaches. First we will test whether reduced LPP3 and/or increased S1P levels will increase muscle cell regeneration in mdx mice using the existing muscle regeneration methods in Seattle Muscular Dystrophy Consortium (2.1). Second, we will take advantage of the induced pluripotent stem cell (iPSC) expertise in my laboratory to generate skeletal myoblasts using iPSCs derived from muscular dystrophy patients and test whether the regeneration capacity of these myoblasts is increased due to the modified levels of LPP3 and/or S1P (2.2). This challenge grant combines multiple fields; two Seattle based consortiums and three laboratories. The work from this combination of disciplines is bound to generate new exciting advances in muscular dystrophy therapy The goal of this grant is to generate a potential therapy for muscular dystrophy using a lipid phosphate phosphatase, Wunen/LPP3. Reduction of this particular lipid phosphatase shows partial rescue of defects seen in a Drosophila muscular dystrophy model. We will now test whether reduction of this phosphatase, LPP3 can suppress muscle defects in multiple muscular dystrophy model systems. Ultimately, in the future we wish to test whether manipulating sphingolipid metabolism can serve therapeutic function in muscular dystrophy treatment.

描述(由申请人提供)：本申请涉及广泛的挑战领域(11)再生医学和特殊挑战主题11-AR-101：肌肉骨骼和皮肤组织再生。这笔赠款的重点是开发肌肉营养不良治疗的新战略或/和加强现有战略。这项资助是基于一项新的发现，该发现表明，LPP3的减少降低了活跃的第二信使S1P的水平，可以抑制果蝇营养不良的表型。我们现在将研究鞘磷脂途径和dystrophin相互作用的机制，并进一步测试基于wunen/LPP3的策略来改善营养不良小鼠的肌肉功能。肌营养不良症是一种肌肉萎缩的疾病，目前还没有治愈的方法。如前所述，我们的研究已经为这种疾病的治疗提供了一条可能的新途径，它涉及到乌能。我们将详尽地描述Wunen，一种在我们之前的正向遗传筛查中分离出来的抑制物，用于Dystrophin相关糖蛋白复合体突变表型的修饰。Wunen是人类的同源物这项资助是基于一项新的发现，LPP3的减少降低了活跃的第二信使S1P的水平，S1P可以抑制果蝇营养不良的表型。我们现在将研究鞘磷脂途径和dystrophin相互作用的机制，并进一步测试基于wunen/LPP3的策略来改善营养不良小鼠的肌肉功能。肌营养不良症是一种肌肉萎缩的疾病，目前还没有治愈的方法。如上所述，我们的研究已经为这种疾病的治疗提供了一条可能的新途径，它涉及到wunen(LPP3)，许多研究表明它参与了生物活性脂质水平的调节，特别是鞘氨醇1-磷酸(S1P)。S1P与前成肌细胞(成肌干细胞)的维持以及成肌细胞向肌管的分化有关。通过改变鞘脂信号来抑制肌营养不良表型的肌肉再生策略的可能性很高，具有非常有效的潜力。我们将确定五能是否通过S1P抑制细胞凋亡和/或通过促进成体肌肉前体细胞的增殖或促进成体肌肉卫星干细胞的增殖来增加肌肉生成，这在果蝇中尚未确定。我们将确定肌肉卫星干细胞是否存在于哺乳动物肌肉卫星干细胞相同的生理生态位中，即在现有肌肉纤维的基膜和肌膜之间。总之，我们将建立和扩展神经鞘脂信号在果蝇肌肉形成中的作用，并确定缓解营养不良果蝇疾病表型的机制。此外，我们将测试Wunen/LPP3的减少是否有利于肌肉营养不良的哺乳动物动物模型，特别是MDX小鼠(以及未来的狗模型)。我们希望在这个模型中产生再生治疗策略来缓解疾病的表型。这些策略将包括减少LPP3功能以及增加其下游效应子S1P的存在。我们认为，增加鞘磷脂信号将增加肌肉干细胞和成肌细胞的再生潜力，从而提高肌营养不良症的治疗效果。我们将通过两种不同的方法来检验这一假设。首先，我们将使用西雅图肌肉营养不良协会现有的肌肉再生方法来测试降低LPP3和/或增加S1P水平是否会增加MDX小鼠的肌肉细胞再生(2.1)。其次，我们将利用我实验室的诱导多能干细胞(IPSC)专业知识，使用来自肌营养不良患者的IPSC来生成骨骼肌成肌细胞，并测试这些成肌细胞的再生能力是否因修改LPP3和/或S1P(2.2)的水平而增加。这项挑战拨款结合了多个领域；两个总部位于西雅图的财团和三个实验室。这一结合学科的工作必将在肌营养不良症治疗方面产生新的令人兴奋的进展。这笔赠款的目标是利用一种脂质磷酸磷酸酶Wunen/LPP3产生一种治疗肌营养不良症的潜在疗法。这种特殊的脂质磷酸酶的减少表明，果蝇肌营养不良模型中的缺陷得到了部分修复。我们现在将在多发性肌营养不良症模型系统中测试这种磷酸酶LPP3的减少是否可以抑制肌肉缺陷。最终，在未来，我们希望测试操控鞘磷脂代谢是否可以在肌营养不良症的治疗中发挥治疗作用。