Establishment of stable zebrafish genetic models for studying myotonic dystrophy

建立用于研究强直性肌营养不良的稳定斑马鱼遗传模型

• 批准号: 9328830
• 负责人: Melissa N Hinman
• 金额: $ 5.71万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9328830
• 关键词: Abdominal Pain; Affect; Alleles; Alternative Splicing; Animal Model; Animals; Bacteria; Binding; Biological Models; CRISPR/Cas technology; Cell Line; Cell model; Cells; Constipation; Data; Defect; Diarrhea; Digestion; Disease; Disease model; Etiology; Event; Fertility; Fishes; Functional disorder; Future; Gastric Emptying; Gastrointestinal tract structure; Genes; Genetic Models; Genetic Transcription; Germ-Free; Goals; Human; Human Genetics; Image; Individual; Intestinal Motility; Intestinal Pseudo-Obstruction; Intestines; Larva; Lead; Length; Link; Measures; Microscopy; Modeling; Molecular; Molecular Target; Morphology; Movement; Mus; Muscle; Muscle Weakness; Muscular Atrophy; Muscular Dystrophies; Mutate; Mutation; Myotonia; Myotonic Disorders; Myotonic Dystrophy; Nucleotides; Pathology; Patients; Pharmaceutical Preparations; Phenotype; Physiology; Play; Preclinical Drug Evaluation; Process; Protein Family; Proteins; RNA; RNA Splicing; RNA-Binding Proteins; Role; Series; Severities; Severity of illness; Skeletal Muscle; Study models; Symptoms; System; Testing; Tissues; Transgenic Organisms; Visceral; Work; Zebrafish; body system; cell motility; disease phenotype; drug candidate; experience; gastrointestinal system; gut microbiota; human disease; improved; insight; loss of function; microbiota; motility disorder; mouse model; mutant; novel; offspring; overexpression; prevent; protein function; screening; small molecule; tool; transcriptome sequencing

Project Summary/Abstract Myotonic dystrophy (DM) is one of the most common forms of muscular dystrophy, affecting 1 in 8,000 people. It is common for DM patients to have digestive symptoms, often involving altered gut movement, including slowed stomach emptying, slowed intestinal movement, abdominal pain, diarrhea, and constipation. Intestinal bacteria play important roles in digestion, and many DM patients have overgrowth of these bacteria. A quarter of DM patients describe digestive symptoms as the most disabling aspect of the disease, yet these symptoms are poorly understood. The proposed work uses zebrafish to study DM because digestive symptoms can be observed directly in live, transparent larvae, and because zebrafish produce large numbers of offspring that can be used to screen for drugs to improve DM symptoms. The most common form of DM is caused by expression of RNA containing long repeats of CUG nucleotides, which bind to and sequester the MBNL family of proteins and prevent them from regulating an important molecular process called alternative splicing. Zebrafish have been generated that mimic the loss of MBNL protein function in DM by mutation of mbnl genes, and these fish show some of the same alternative splicing changes that are seen in human disease. Fish are also being made that mimic DM in another way, by expressing CUG repeat RNA. Using the new DM model fish, the first goal is to determine whether they have DM-like symptoms and to identify molecular changes that contribute to these symptoms. The next goal is to measure changes in gut movement in live zebrafish DM models using microscopy, to determine whether bacteria contribute to these changes, and to determine which tissue types are important for these symptoms. Finally, a small-scale test of candidate drugs to ameliorate the DM-like symptoms of model fish will be performed. This will serve as a proof-of-principle for future large-scale screening for drugs that improve DM-like symptoms in fish with the long term goal of identifying drugs to reduce or eliminate symptoms in DM patients. Overall, these studies will provide important insight into the poorly understood digestive symptoms of DM and the molecular mechanisms behind them. They will also generate novel animal model tools that will be invaluable for studying other aspects of DM pathology and for screening for drugs to improve the lives of individuals with DM. !

项目摘要/摘要 强直性肌营养不良症(DM)是最常见的肌营养不良症之一， 每8,000人中就有1人受到影响。糖尿病患者通常会有消化症状，通常 涉及肠道运动的改变，包括胃排空减慢，肠道减慢 运动、腹痛、腹泻和便秘。肠道细菌扮演着重要的角色 在消化方面，许多糖尿病患者都有这些细菌的过度生长。四分之一的DM 患者将消化症状描述为该病最令人无力的方面，但这些 人们对这些症状知之甚少。这项拟议的工作使用斑马鱼来研究DM，因为 消化症状可以直接在活的透明幼虫身上观察到，因为斑马鱼 产生大量后代，可用于筛选改善糖尿病的药物 症状。 DM最常见的形式是由含有长重复序列的RNA表达引起的 CUG核苷酸，它与MBNL蛋白家族结合并隔离，并防止它们 调节一种叫做选择性剪接的重要分子过程。斑马鱼已经被 通过MBNL基因突变产生模拟糖尿病患者MBNL蛋白功能丧失的基因，以及 这些鱼表现出一些与人类疾病相同的选择性剪接变化。 鱼类也在以另一种方式模仿DM，通过表达CUG重复RNA。 使用新的DM模型鱼，第一个目标是确定它们是否有类似DM的鱼 并确定导致这些症状的分子变化。下一个目标 是使用显微镜测量活斑马鱼DM模型中肠道运动的变化，以 确定细菌是否导致了这些变化，并确定了哪些组织类型 对这些症状很重要。最后，对候选药物进行小规模测试以改善 将执行模型鱼的DM样症状。这将作为以下原则的证明 未来大规模筛选可长期改善鱼类类糖尿病症状的药物 确定减轻或消除糖尿病患者症状的药物的目标。 总体而言，这些研究将为了解鲜为人知的消化系统疾病提供重要的见解。 糖尿病的症状及其背后的分子机制。它们还将产生小说 动物模型工具将对研究DM病理学的其他方面和 筛选药物以改善糖尿病患者的生活。 好了！