Generation and characterization of a Cre-Lox regulated transgenic zebrafish model of SBMA

Cre-Lox 调节的 SBMA 转基因斑马鱼模型的生成和表征

基本信息

批准号：
10784254
负责人：
Heather L Montie
金额：
$ 15.16万
依托单位：
PHILADELPHIA COLLEGE OF OSTEOPATHIC MED
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-11 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10784254
关键词：
AR gene Actins Adult Affect Androgen Receptor Androgens Animals Biochemical Biological Assay Biology Birth Breeding CAG repeat Chemicals Clinical Trials Design Collaborations Complementary DNA Cre lox recombination system Cre-LoxP DNA Binding DNA cassette Data Dedications Dependence Disease DsRed Exhibits Female Functional disorder Funding Future Generations Genetic Genetic Recombination Genotype Glutamine Goals Heart Histology Hormone Receptor Human In Vitro Kennedy Syndrome Laboratories Length Libraries Ligand Binding Domain Ligands Limb structure Link LoxP-flanked allele Maintenance Microscopy Modeling Modification Molecular Motor Motor Activity Motor Neurons Movement Mus Muscle Weakness Muscular Atrophy National Institute of Neurological Disorders and Stroke Nerve Degeneration Neuroanatomy Neurodegenerative Disorders Neuromuscular Diseases Partner in relationship Pathogenesis Pathology Patients Pharmaceutical Preparations Phenotype Process Production Property Proteins Research Research Personnel Rodent Model Role Science Site Skeletal Muscle Specificity Speed Stanolone Structure Study models Swimming System Terminator Codon Therapeutic Therapeutic Intervention Time Tissues Transgenes Transgenic Organisms Translating Validation Vertebrates Zebrafish cell type cohort cost cost comparison human disease in vivo inducible Cre male men model organism molecular pathology motor deficit motor disorder mouse model muscle physiology neuromuscular patient population pharmacologic polyglutamine promoter receptor receptor expression receptor function spinal and bulbar muscular atrophy therapeutic development therapeutic target transgene expression

项目摘要

Spinal and bulbar muscular atrophy (SBMA or Kennedy's disease) is a slowly progressive, X-linked neuromuscular disease affecting men. It is caused by the expansion of a CAG repeat within the androgen receptor (AR) gene, encoding a glutamine tract in the protein. With no cure or therapy, identifying potential therapeutic interventions for this patient population is pressing. We aim to create a Cre-inducible, low cost, and high-throughput model of SBMA using zebrafish. Discoveries made from zebrafish research are highly translatable to humans since zebrafish are vertebrates with extensive structural homology to human neuroanatomy and muscle physiology. More specifically for SBMA patients, zebrafish express an AR that is highly homologous in structure and function to human AR. Cre-inducibility of the human AR transgene will enable the production of founder lines that are able to express high levels of the toxic polyglutamine-expanded AR. We hypothesize this will lead to robust motor dysfunction in larval zebrafish. These larval zebrafish modeling SBMA will be amenable to quick (over 1-2 weeks) chemical and drug library screens, or genetic modification(s) to identify modulators of motor function. Such assays could be set up within only a few days and at a very low cost compared to such assays in rodent models. Moreover, tissue specific Cre zebrafish lines can be crossed with these SBMA zebrafish in future studies to investigate the relative role of motor neurons verses skeletal muscle in the onset and progression of SBMA pathogenesis. A better understanding of the relative roles of each of these cell types in disease will support clinical trial design for SBMA patients. Further, because SBMA shares properties with other neurodegenerative, neuromuscular, and polyglutamine diseases, therapeutic targets identified in the SBMA zebrafish model may also be applicable to some of these other disorders.

脊髓和延髓肌萎缩症（SBMA 或肯尼迪病）是一种缓慢进展的 X 连锁疾病影响男性的神经肌肉疾病。它是由雄激素内 CAG 重复序列的扩展引起的受体（AR）基因，编码蛋白质中的谷氨酰胺束。在没有治愈方法或疗法的情况下，识别潜在的针对这一患者群体的治疗干预措施刻不容缓。我们的目标是创造一种可诱导的、低成本的、使用斑马鱼的 SBMA 高通量模型。斑马鱼研究的发现受到高度评价可转化为人类，因为斑马鱼是与人类具有广泛结构同源性的脊椎动物神经解剖学和肌肉生理学。更具体地说，对于 SBMA 患者，斑马鱼表达的 AR 是在结构和功能上与人类AR高度同源。人类 AR 转基因的诱导能力将使能够表达高水平的有毒聚谷氨酰胺扩增 AR 的创始系的生产。我们假设这将导致斑马鱼幼虫出现强烈的运动功能障碍。这些幼虫斑马鱼建模SBMA 将能够进行快速（超过 1-2 周）化学和药物库筛选，或进行基因改造识别运动功能的调节器。这种检测只需几天就能完成，而且成本非常低与啮齿动物模型中的此类测定进行比较。此外，组织特异性 Cre 斑马鱼系可以与这些 SBMA 斑马鱼将在未来的研究中研究运动神经元与骨骼肌的相对作用 SBMA 发病机制的发生和进展。更好地理解其中每一个的相对作用疾病中的细胞类型将支持 SBMA 患者的临床试验设计。此外，由于 SBMA 共享财产与其他神经退行性疾病、神经肌肉疾病和多聚谷氨酰胺疾病一样，在 SBMA 斑马鱼模型也可能适用于其中一些其他疾病。