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

• 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
• 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

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.

脊柱和鳞茎肌肉萎缩（SBMA或肯尼迪氏病）是一种缓慢的X连锁 神经肌肉疾病影响男性。它是由雄激素内CAG重复膨胀引起的 受体（AR）基因，编码蛋白质中的谷氨酰胺道。没有治愈或治疗，确定潜力 该患者人群的治疗干预措施正在紧迫。我们旨在创建一个可诱导的，低成本，并且 使用斑马鱼的SBMA高通量模型。斑马鱼研究的发现很高 可以翻译为人类，因为斑马鱼是脊椎动物，与人类具有广泛的结构同源性 神经解剖学和肌肉生理。斑马鱼更具体地适用于SBMA患者 与人类AR的结构和功能高度同源。人类AR转基因的Cre-Cre-odibens将启用 能够表达高水平有毒多谷氨酰胺膨胀AR的创始人线的产生。我们 假设这将导致幼虫斑马鱼中强大的运动功能障碍。这些幼虫斑马鱼建模SBMA 将适合快速（超过1-2周）化学和药物库筛选，或者遗传修饰 识别运动功能的调节器。这样的测定可以在短短几天内进行，并且成本非常低 与啮齿动物模型中的这种测定相比。此外，可以与组织特异性的斑马鱼线交叉 这些SBMA斑马鱼在未来的研究中研究了运动神经元的相对作用，骨骼肌 在SBMA发病机理的发作和进展中。更好地理解每个这些的相对作用 疾病的细胞类型将支持SBMA患者的临床试验设计。此外，由于SBMA共享属性 使用其他神经退行性，神经肌肉和多谷氨酰胺疾病，在 SBMA斑马鱼模型也可能适用于其他一些疾病。