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Targeting vascular endothelium for muscular dystrophy therapy

靶向血管内皮治疗肌营养不良症

基本信息

批准号：
10379330
负责人：
ATSUSHI ASAKURA
金额：
$ 16.88万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10379330
关键词：
3-Dimensional Address Affinity Anatomy Angiogenic Factor Binding Blood Blood Vessels Blood capillaries Capillary Endothelial Cell Cell Count Cell Differentiation process Cell Maintenance Characteristics Chronic Complex Data Defect Development Disease Duchenne muscular dystrophy Dystrophin Endothelial Cells Endothelium Equilibrium Fibroblast Growth Factor Fibrosis Gene Expression Generations Genes Goals Growth Histologic Homeostasis Human Image Immunization Immunize Impairment Knockout Mice Lead Ligands Llama Maintenance Membrane Mesenchymal Modeling Molecular Monoclonal Antibodies Mus Muscle Muscle Contraction Muscle Fibers Muscle Weakness Muscle function Muscle satellite cell Muscular Dystrophies Mutation Myofibroblast Myopathy Natural regeneration Notch Signaling Pathway Paracrine Communication Pathogenesis Pathologic Pathology Pathway interactions Patients Phage Display Phenotype Population Process Proteins Protocols documentation Quality of life Regenerative capacity Reporter Resolution Role Serum Severities Skeletal Muscle Snails Structure Techniques Technology Therapeutic Time Tissues Translating VEGFA gene Vascular Endothelial Cell Vascular Endothelial Growth Factors Vascular Endothelium Work aged angiogenesis cell growth cell type conditional knockout density functional improvement improved knockout gene mdx mouse muscle regeneration myogenesis nanobodies notch protein novel novel therapeutics postnatal prevent receptor recruit repaired satellite cell self-renewal skeletal muscle wasting stem cell population stem cells therapeutic target transcription factor

项目摘要

ABSTRACT Skeletal muscle is a highly ordered, yet complex tissue which contains several cell types that interact with each other to maintain structure and homeostasis. Muscle satellite cells (SCs) are a stem cell population responsible for muscle growth, repair and regeneration. Maintenance of the balance between SC differentiation and self- renewal is required for muscle homeostasis. A defect in self-renewal ability leads to a decrease in SC number, resulting in depletion of the SC pool and reduced muscle regeneration capacity, which occurs in aged and diseased muscle, including Duchenne muscular dystrophy (DMD). DMD is a progressive disorder in which the absence of the dystrophin protein results in loss of the dystrophin bridge at the muscle membrane. Recent work demonstrates the angiogenic impairment of the ECs in DMD model mdx mice. We demonstrates the increased vascular density can ameliorate phenotypes associated with DMD. However, the importance of angiogenesis in DMD treatment has not yet been well addressed. A recent study demonstrated that SCs are preferentially located next to capillary endothelial cells (ECs). However, the exact relationship between SCs and ECs has yet to be examined. Recently, we established an optimized tissue clearing protocol for skeletal muscle. We utilized fluorescent reporters for SCs and ECs along with tissue clearing to demonstrate the close proximity of SCs to capillaries in 3-dimentional imaging, suggesting the juxtavascular niche of SCs for stem cell maintenance. SCs express vascular endothelial cell growth factor (VEGF) and Notch receptors which is dynamically altered during the regeneration process. Therefore, we hypothesize that SCs recruit capillary ECs via VEGF to establish a juxtavascular niche for SC maintenance during homeostasis and regeneration. In this proposal, we will determine the extent to which an increased vascular niche in postnatal conditional knockout of Flt1 and by blocking Flt1 via anti-Flt1 nanobodies, created by a combination of llama-immunization and phage display technologies, in mdx mice can increase SC number, reduced pathological endothelial-to-mesenchymal transition (EndMT), and ameliorate phenotypes associated with DMD. Consequently, we will elucidate key molecules that regulate angio- myogenesis, including those involved in SC homeostasis, which is fundamental to developing therapeutic approaches to treat DMD. SCs and their juxtavascular niche are potential therapeutic targets to induce and maintain SC populations that slow the loss of skeletal muscle function with DMD.

抽象的骨骼肌是一种高度有序但复杂的组织，包含多种细胞类型，每种细胞类型相互作用其他维持结构和体内平衡。肌肉卫星细胞 (SC) 是干细胞群，负责用于肌肉生长、修复和再生。维持SC分化和自我分化之间的平衡肌肉稳态需要更新。自我更新能力缺陷导致SC数量减少，导致 SC 池耗尽并降低肌肉再生能力，这种情况发生在老年人和老年人中患病肌肉，包括杜氏肌营养不良症 (DMD)。 DMD 是一种进行性疾病，其中肌营养不良蛋白的缺失会导致肌膜上肌营养不良蛋白桥的丢失。最近的工作证明 DMD 模型 mdx 小鼠 EC 的血管生成受损。我们展示了增加的血管密度可以改善与 DMD 相关的表型。然而，血管生成的重要性 DMD 的治疗尚未得到很好的解决。最近的一项研究表明，SC 优先位于紧邻毛细血管内皮细胞（EC）。然而，SC 和 EC 之间的确切关系尚未确定。检查了。最近，我们建立了一种优化的骨骼肌组织清除方案。我们利用了 SC 和 EC 的荧光报告基因以及组织透明化，以证明 SC 与三维成像中的毛细血管，表明干细胞维持的血管旁利基。 SC 表达血管内皮细胞生长因子 (VEGF) 和 Notch 受体，这些受体在生长过程中动态变化再生过程。因此，我们假设 SCs 通过 VEGF 招募毛细血管 ECs 来建立稳态和再生过程中 SC 维持的近血管生态位。在本提案中，我们将确定出生后条件性敲除 Flt1 并通过阻断 Flt1 来增加血管生态位的程度抗 Flt1 纳米抗体，通过结合美洲驼免疫和噬菌体展示技术在 mdx 中创建小鼠可以增加 SC 数量，减少病理性内皮间质转化 (EndMT)，改善与 DMD 相关的表型。因此，我们将阐明调节血管的关键分子肌生成，包括那些参与 SC 稳态的肌生成，这是开发治疗药物的基础治疗 DMD 的方法。 SC 及其血管旁生态位是诱导和治疗的潜在治疗靶点维持 SC 群体，减缓 DMD 导致的骨骼肌功能丧失。