Establishing an FSHD-like mouse for therapeutic development

建立类 FSHD 小鼠用于治疗开发

• 批准号: 9167305
• 负责人: Peter L Jones
• 金额: $ 1.74万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9167305
• 关键词: Address; Adult; Affect; Alternative Splicing; Animal Model; Apoptosis; Binding; Biological Markers; Breathing; Canis familiaris; Characteristics; Child; Chromatin Structure; Clinical; Comb animal structure; D4Z4; DNA; DNA Binding; DNA Methylation; DNA-Binding Proteins; Data; Development; Disease; Disease Progression; Elements; Engineering; Environmental Risk Factor; Epigenetic Process; Event; Exhibits; Facioscapulohumeral Muscular Dystrophy; Female; Functional disorder; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Structure; Generations; Genes; Genetic; Genetic Recombination; Goals; Hair; Hand Strength; Histopathology; Homeodomain Proteins; Hormones; Human; In Vitro; Inflammatory Response; Lead; Life; Live Birth; Mediating; Mediator of activation protein; Messenger RNA; Modeling; Molecular Profiling; Mus; Muscle; Muscle Weakness; Muscle function; Muscular Dystrophies; Myopathy; Nonsense-Mediated Decay; Oxidative Stress; Pathology; Pathway interactions; Patients; Phenotype; Preclinical Testing; Proteins; RNA Splicing; Regulation; Resources; Running; Severities; Skeletal Muscle; Stem cells; Tamoxifen; Testis; Therapeutic; Transgenic Mice; WNT Signaling Pathway; Walking; Wheelchairs; base; career; cytotoxicity; design; epigenetic regulation; exhaust; histone modification; male; model development; mouse model; multicatalytic endopeptidase complex; muscle strength; muscular dystrophy mouse model; myogenesis; pathogenic isoform; prevent; protein aggregation; repaired; research clinical testing; targeted treatment; therapeutic development; therapeutic target; tool; transcription factor

PROJECT SUMMARY Facioscapulohumeral muscular dystrophy (FSHD) is the most prevalent myopathy afflicting males and females, children and adults. In the majority of clinical FSHD cases, muscle weakness is not noticeable until the second or third decade of life followed by a progressive pathology impacting many facets of everyday life, ranging from being unable to comb ones own hair or walk the dog to including having to change or abandon careers, loss of independence and, in ~20% of FSHD patients, becoming wheelchair bound an/or require aid in breathing. Currently there are no treatments to slow down, stop, or reverse disease progression. A major impediment to developing ameliorative treatments is the lack of a reliable phenotypic FSHD-like animal model based on expression of the DUX4 gene, widely considered the key mediator of FSHD pathophysiology. This project directly addresses this void. In FSHD, increased expression of DUX4-fl, which can function as a DNA-binding transcription factor, alters the gene expression profiles of muscles and initiates a cascade of events ultimately leading to FSHD pathophysiology. Thus, the DUX4-fl mRNA, DUX4- FL protein and downstream targets are all excellent targets for therapeutic development. For this project, we have successfully engineered a line of transgenic mice that contains the human DUX4- fl gene maintaining its native gene structure. These mice are validated to express correctly spliced DUX4-fl mRNA upon Cre-mediated recombination and develop an aberrant muscle phenotype when DUX4-fl is expressed in developing muscle. This data strongly supports the ultimate generation of an FSHD-like model mouse; however, the precise expression conditions that will result in a useful FSHD-like phenotype are still not known. Here, we will use a commercially available line of mice expressing tamoxifen-inducible Cre in skeletal muscles to determine the conditions required to generate reproducible FSHD-like phenotypes over a range of severities. A successful FSHD-like mouse will appear initially healthy, then develop a progressive myopathic phenotype based on quantifiable metrics of muscle strength and function including grip strength, rotarod, and the maximum distance each is capable of running before becoming exhausted. The phenotypes will be confirmed by FSHD-like gene expression analysis and histopathology. Completion of this project will provide the FSHD field with valuable tools for better understanding FSHD pathogenic progression and mechanisms. In addition, these models will serve as a resource for pre-clinical testing of therapeutic strategies targeting the DUX4-fl mRNA and protein as ameliorative treatments for FSHD.

项目摘要 面肩肱型肌营养不良症（FSHD）是困扰男性的最普遍的肌病， 女性、儿童和成人。在大多数临床FSHD病例中，肌肉无力并不是 直到生命的第二个或第三个十年才明显，随后是影响许多人的进行性病理学 日常生活的方方面面，从不能自己梳头或遛狗， 不得不改变或放弃职业，失去独立性，在约20%的FSHD患者中， 坐轮椅和/或需要呼吸帮助。目前还没有治疗方法 减缓、停止或逆转疾病进展。发展改良的主要障碍 缺乏基于DUX4表达的可靠的表型FSHD样动物模型 基因，被广泛认为是FSHD病理生理学的关键介质。该项目直接解决了这一问题 虚空在FSHD中，DUX4-fl的表达增加，DUX4-fl可以作为DNA结合转录因子发挥作用。 因子，改变肌肉的基因表达谱，并启动一系列事件，最终导致 FSHD的病理生理学因此，DUX4-fl mRNA、DUX4-FL蛋白和下游靶标都是靶向的。 治疗开发的绝佳目标。在这个项目中，我们成功地设计了一条生产线， 含有人DUX4-fl基因的转基因小鼠保持其天然基因结构。这些 经验证，小鼠在Cre介导的重组后表达正确剪接的DUX4-f1 mRNA， 当DUX4-f1在发育中的肌肉中表达时，发育异常的肌肉表型。这 数据强烈支持FSHD样模型小鼠的最终生成;然而， 产生有用的FSHD样表型的表达条件仍然未知。这里我们 将使用在骨骼肌中表达他莫昔芬诱导型Cre的市售小鼠系， 确定在一定范围内产生可再现的FSHD样表型所需的条件， 严重性一个成功的FSHD样小鼠将出现最初的健康，然后发展为渐进的 基于肌肉力量和功能（包括握力）的可量化指标的肌病表型 力量，旋转棒，以及每个人在筋疲力尽之前能够跑的最大距离。 将通过FSHD样基因表达分析和组织病理学确认表型。 该项目的完成将为FSHD领域提供有价值的工具，以便更好地了解 FSHD致病进展和机制。此外，这些模型将作为 靶向DUX4-fl mRNA和蛋白质的治疗策略的临床前测试资源， FSHD的改善治疗。