Unveiling the molecular mechanisms in TPM3-related myopathy and therapies

揭示 TPM3 相关肌病的分子机制和治疗方法

• 批准号: 10670411
• 负责人: Matthias Lambert
• 金额: $ 10.62万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-29 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10670411
• 关键词: Acceleration; Address; Affect; Alleles; Animal Model; Atrophic; Biological Assay; Biological Models; Biology; Calcium; Cell Nucleus; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Data; Data Analyses; Development; Disease; Drug Modelings; Drug Screening; Fast-Twitch Muscle Fibers; Fiber; Functional disorder; Funding; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Hereditary Disease; Histology; Hypertrophy; In Vitro; Knowledge; Libraries; Mentors; Microfilaments; Missense Mutation; Modeling; Molecular; Mus; Muscle; Muscle Development; Muscle Weakness; Muscle function; Muscle satellite cell; Muscular Atrophy; Mutation; Myopathy; Neuromuscular Diseases; Pathogenesis; Pathology; Pathway interactions; Patients; Performance; Phase; Phenotype; Point Mutation; Population; Positioning Attribute; Procedures; Prognosis; Quality of life; RNA Interference; Regulatory Pathway; Research; Research Personnel; Severity of illness; Skeletal Muscle; Slow-Twitch Muscle Fibers; Small Interfering RNA; Swimming; Testing; Therapeutic; Tissues; Validation; Zebrafish; autosome; career development; congenital myopathy; design; efficacy evaluation; efficacy testing; experience; experimental study; fascinate; humanized mouse; improved; in vivo; insight; interest; large scale data; mouse model; muscle metabolism; mutant; professor; single nucleus RNA-sequencing; skills; small hairpin RNA; tenure track; therapeutic development; tool

Project Summary Autosomal dominant TPM3-related myopathy is primarily caused by missense mutations in the TPM3 gene that lead to muscle atrophy and weakness, and substantially affect patient’s quality of life. So far, there is no treatment for TPM3-related myopathy. K99 mentored phase: Currently, the absence of representative animal model systems limits the emergence of new pathophysiological findings in TPM3-related myopathy as well as potential therapeutics. To overcome this gap, we have recently generated CRISPR-based Tpm3 mice that carry one of the most prevalent mutations seen in patients. Our objectives are (1) to study the muscle pathology in the humanized mouse model of TPM3-related myopathy, and (2) to dissect the transcriptional changes in slow and fast myonuclear populations. Furthermore, we have characterized a tpm3-deficient zebrafish that show similar pathophysiological features of myopathy and holds promise for the development of complementary approaches that would be inconceivable with mammalian model systems. We also aim (3) to generate CRISPR-based tpm3 zebrafish that carry the same most prevalent mutation seen in patients. R00 independent phase: Zebrafish have become a powerful model to accelerate the discovery of potential therapeutics. Our objective is (4) to develop a drug screening assay in tpm3 zebrafish. Allele-specific RNA interference has also emerged as a powerful strategy for dominant inherited diseases such as TPM3-related myopathy. Based on our preliminary data, TPM3 haploinsufficiency is not the likely underlying mechanism of the myopathy. To complement our study in zebrafish, we aim (5) to screen for siRNAs that suppress the mutant allele expression without reducing the wild-type allele in in vitro experiments and to test their efficacy in vivo in our Tpm3 mouse. My long-term goal is to become an independent investigator with a lab that combines the genetic and molecular tools to study the neuromuscular diseases and develop therapeutic approaches. During the K99 mentored phase, I will gain additional skills in conceptual, technical and career development aspects which will enable me to make a successful transition to an independent position with my own research group. My short-term goals are (1) to acquire further skills in experimental procedures to study the muscle pathology in mouse and zebrafish, (2) to gain expertise in large-scale data analysis, (3) to improve my knowledge in the design of therapeutic strategies for neuromuscular disorders, (4) to obtain an independent tenure-track assistant professor position, and (5) to successfully obtain R01 funding within 5 years of this proposal.

项目摘要 常染色体显性TPM3相关肌病主要由TPM3基因的错义突变引起 这导致肌肉萎缩和虚弱，并严重影响患者生活质量。到目前为止还没有 治疗TPM3相关肌病。 K99指导阶段：目前，缺乏代表性的动物模型系统限制了K99的出现。 TPM3相关肌病的新病理生理学发现以及潜在的治疗方法。为了克服这个 差距，我们最近培育了基于CRISPR的Tpm 3小鼠，它们携带最普遍的突变之一 在病人身上看到的。我们的目的是（1）研究人源化小鼠模型中的肌肉病理学， TPM3相关的肌病，和（2）解剖慢和快肌病的转录变化， 人口。此外，我们已经表征了tpm3缺陷的斑马鱼，其表现出类似的 肌病的病理生理学特征，并有望发展互补的 这些方法在哺乳动物模型系统中是不可想象的。我们的目标是（3）生成 基于CRISPR的tpm3斑马鱼携带与患者相同的最常见突变。 R00独立阶段：斑马鱼已成为加速发现潜力的强大模式 治疗学我们的目标是（4）建立一种在tpm3斑马鱼中进行药物筛选的方法。等位基因特异性RNA 干扰也已成为显性遗传性疾病（如TPM3相关疾病）的一种强有力的策略。 肌病根据我们的初步数据，TPM3单倍不足不是TPM3的可能潜在机制。 肌病为了补充我们在斑马鱼中的研究，我们的目标是（5）筛选抑制 在体外实验中不减少野生型等位基因的突变等位基因表达，并测试它们在 在我们的Tpm 3小鼠体内。 我的长期目标是成为一名独立的调查员，拥有一个结合基因和 研究神经肌肉疾病和开发治疗方法的分子工具。在K99 在指导阶段，我将获得概念、技术和职业发展方面的额外技能， 使我能够成功地过渡到我自己的研究小组的独立职位。 我的短期目标是（1）获得进一步的技能，在实验程序，研究肌肉病理学 （2）获得大规模数据分析的专业知识，（3）提高我在 设计神经肌肉疾病的治疗策略，（4）获得独立的终身制 助理教授职位，以及（5）在本提案的5年内成功获得R01资助。