Investigating the role of the conserved transcriptional regulator Apterous on muscle development

研究保守转录调节因子 Apterous 对肌肉发育的作用

• 批准号: 10377502
• 负责人: Krista Carol Dobi
• 金额: $ 15.32万
• 依托单位: BERNARD M. BARUCH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10377502
• 关键词: Abdominal Muscles; Adopted; Adult; Affect; Aging; Animal Model; Area; Cellular Morphology; Cellular biology; Cessation of life; DNA Binding; Data; Deposition; Development; Disease; Dorsal; Drosophila genus; Drosophila melanogaster; Embryo; Eye; Failure; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; Goals; Health; Heart Diseases; Homeobox; Human; Image; Immunohistochemistry; In Situ Hybridization; Integrins; Kidney Failure; Knowledge; Lateral; Lead; Learning; Limb structure; Link; Location; Lung diseases; Malignant Neoplasms; Microscopy; Modeling; Morbidity - disease rate; Morphogenesis; Morphology; Muscle; Muscle Cells; Muscle Development; Muscular Atrophy; Muscular Dystrophies; Myoblasts; Myopathy; Only Child; Orthologous Gene; Patients; Pattern; Physiology; Positioning Attribute; Property; Proteins; Quantitative Reverse Transcriptase PCR; Regulation; Regulator Genes; Reporter; Research; Role; Sarcomeres; Shapes; Specific qualifier value; Spinal Muscular Atrophy; Stem Cell Development; Structural Protein; System; Talin; Tendon structure; Testing; To specify; Transgenes; Validation; Vinculin; Work; age effect; base; experimental study; fly; gene network; homeodomain; loss of function; loss of function mutation; mortality; muscle form; mutant; nerve supply; prevent; programs; side effect; stem cell therapy; stem cells; targeted treatment; therapy development; transcription factor; transcriptome sequencing

PROJECT SUMMARY There is a critical need to develop treatments for muscle wasting, which increases morbidity and mortality of patients suffering from muscular dystrophies, myopathies, cancer, kidney failure and pulmonary disease. Stem cell therapies require the ability to generate muscles of particular sizes and shapes to replace damaged muscles in, for example, the round eye or the elongated limb. A thorough understanding of how muscles with specific properties develop will allow us to "program" muscle cells to adopt specific properties, a key step towards developing these kinds of treatments. The long term goal of the proposed work is to use the fruit fly, Drosophila melanogaster, to determine the cellular mechanisms underlying the development of somatic muscles with distinct sizes, shapes, orientations, innervations, attachments and gene expression patterns. Within each muscle these properties are encoded by distinct sets of gene regulatory factors, including the conserved genes Apterous (Ap), Midline (Mid), and Muscle-segment homebox (Msh). We are focused on a subset of embryonic abdominal muscles that express Ap, Mid and Msh. We have found that expression of Ap outside its normal pattern dramatically disrupts the musculature, leading to changes in muscle positioning and loss of muscle attachments. The failure of the muscle-tendon connection prevents these mutant embryos from generating the force needed to hatch from its eggshell and leads to death. Our hypothesis is that Ap directly regulates genes involved in muscle orientation and the selection of direct versus indirect muscle attachment by functioning in a network of gene regulators including Mid and Msh. We will test this hypothesis with three Aims: 1) examining the location and level of muscle guidance and attachment factors in Ap mutant backgrounds; 2) identifying the genes regulated by Ap; and 3) determining the interactions between Ap, Msh and Mid during muscle development. The proposed work combines gene expression analyses with cell biology, microscopy and genetics to learn how muscle connections are made and determine how many genes are targets of Ap regulation. Taken together, the experiments described in this proposal will determine how muscle properties like shape, orientation and attachment type are specified, which will inform our understanding of the vertebrate orthologs of muscle development in humans and lead to the development of therapies.

项目摘要 迫切需要开发肌肉浪费的治疗方法，这增加了发病率和死亡率 患有肌肉营养不良，肌病，癌症，肾衰竭和肺部疾病的患者。干 细胞疗法需要能够产生特定尺寸和形状的肌肉以替代受损的肌肉 例如，肌肉在圆眼或细长的肢体中。对肌肉的透彻了解 开发特定的特性将使我们能够“编程”肌肉细胞采用特定特性，这是关键步骤 致力于开发这些治疗方法。拟议工作的长期目标是使用果蝇， 果蝇melanogaster，确定体细胞发展的基础机制 具有不同大小，形状，方向，神经，附着和基因表达模式的肌肉。 在每种肌肉中，这些特性都由不同的基因调节因子组编码 保守的基因（AP），中线（中线）和肌肉段Homebox（MSH）。我们专注于 表达AP，MID和MSH的胚胎腹部肌肉的子集。我们发现AP的表达 超出其正常模式会大大破坏肌肉组织，从而导致肌肉定位和 肌肉附着的丧失。肌肉弯曲连接的失败阻止了这些突变胚胎 产生从蛋壳中孵化并导致死亡所需的力。我们的假设是AP直接 调节涉及肌肉取向的基因，以及选择直接与间接肌肉附着的基因 在包括MID和MSH在内的基因调节器网络中发挥作用。我们将以三个目的检验这一假设： 1）检查AP突变背景中肌肉引导和依恋因子的位置和水平； 2） 识别由AP调节的基因； 3）确定AP，MSH和MID之间的相互作用 肌肉发育。提出的工作将基因表达分析与细胞生物学，显微镜结合在一起 和遗传学以了解如何建立肌肉连接并确定AP的靶标有多少基因 规定。综上所述，本提案中描述的实验将决定肌肉特性 像形状一样，定向和依恋类型，这将为我们对脊椎动物的理解提供信息 人类肌肉发育的直系同源物，导致疗法的发展。