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: 7824591
• 负责人: Hannele RUOHOLA-BAKER
• 金额: $ 46.04万
• 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/7824591
• 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）再生医学和特定的挑战主题11-AR-101：肌肉骨骼和皮肤组织再生。该补助金的重点是开发肌营养不良症治疗的新策略或/和加强现有策略。这项资助是基于一项新的发现，该发现表明减少LPP 3，降低活性第二信使S1 P的水平，可以抑制果蝇的营养不良表型。我们现在将研究鞘脂途径和肌营养不良蛋白相互作用的机制，并进一步测试基于wunen/LPP 3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性疾病，目前还没有治愈。如前所述，我们的研究已经产生了一个可能的新进展，治疗这种疾病，它涉及wunen。我们将详尽的特点wunen，在我们以前的正向遗传筛选抗肌萎缩蛋白相关糖蛋白复合物突变表型的修饰剂分离的抑制剂。Wunen是人类的同源物。这项资助是基于一项新的发现，该发现表明，减少LPP 3，降低活性第二信使S1 P的水平，可以抑制果蝇的营养不良表型。我们现在将研究鞘脂途径和肌营养不良蛋白相互作用的机制，并进一步测试基于wunen/LPP 3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性疾病，目前还没有治愈。如前所述，我们的研究已经为这种疾病的治疗产生了可能的新进展，它涉及wunen（LPP 3），许多研究表明它参与调节生物活性脂质的水平，特别是鞘氨醇1-磷酸（S1 P）。S1 P参与前成肌细胞（成体肌肉干细胞）维持以及成肌细胞分化成肌管。通过改变鞘脂信号传导抑制肌营养不良表型的肌肉再生策略的可能性具有非常富有成效的高潜力。我们将确定Wunen是否通过S1 P抑制细胞凋亡和/或通过增加成体肌肉前体细胞的增殖或增加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与哺乳动物肌肉卫星干细胞相同的物理小生境中，即在现有肌纤维的基膜和肌膜之间。总的来说，我们将建立和扩大鞘脂信号在果蝇肌肉形成中的作用，并确定营养不良果蝇疾病表型的机制。此外，我们将测试Wunen/LPP 3的减少是否可以在肌营养不良症中对该疾病的哺乳动物模型有利，特别是mdx小鼠（以及将来的狗模型）。我们希望在这个模型中产生再生治疗策略来减轻疾病表型。这些策略将包括降低LPP 3功能以及增加其下游效应子S1 P的存在。我们认为，增加鞘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治疗的疗效。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营养不良协会（2.1）中现有的肌肉再生方法，测试降低的LPP 3和/或增加的S1 P水平是否会增加mdx小鼠的肌肉细胞再生。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由于LPP 3和/或S1 P水平的改变而增加（2.2）。这项挑战赠款结合了多个领域;两个西雅图的财团和三个实验室。从这个学科的结合工作必将产生新的令人兴奋的进展，在肌营养不良症的治疗，该补助金的目标是产生一个潜在的治疗肌营养不良症使用脂质磷酸磷酸酶，Wunen/LPP 3。这种特殊的脂质磷酸酶的减少显示了在果蝇肌营养不良模型中看到的缺陷的部分拯救。我们现在将测试这种磷酸酶LPP 3的减少是否可以抑制多发性肌营养不良模型系统中的肌肉缺陷。最终，在未来，我们希望测试操纵鞘脂代谢是否可以在肌营养不良症治疗中发挥治疗作用。