Study on PFN1 Pathobiology Using Rat Models

利用大鼠模型进行 PFN1 病理学研究

• 批准号: 10604337
• 负责人: xugang xia
• 金额: $ 59.59万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604337
• 关键词: Affinity; Amyotrophic Lateral Sclerosis; Animal Model; Animals; Axon; Binding; Biology; Cells; Central Nervous System; Complementary DNA; Denervation; Disease; Disease Progression; Disease model; Exhibits; Gene Expression; Genes; Genetic; Genome; Genomics; Hereditary Disease; Human; Impairment; Inherited; Invertebrates; Knock-in; Knock-out; Link; Longevity; Modeling; Molecular; Molecular Chaperones; Monitor; Motor; Motor Neurons; Mouse Strains; Mus; Mutation; Nerve Degeneration; Nucleotides; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pattern; Phenotype; Physiological; Pilot Projects; Rat Transgene; Rattus; Reporting; Research; Rodent; Study models; Synthetic Genes; Therapeutic Effect; Transgenes; Transgenic Mice; Transgenic Organisms; Treatment Efficacy; Vertebrates; aged; biomarker identification; disease phenotype; disease-causing mutation; effective therapy; gene function; human disease; human model; mutant; nervous system disorder; neuron loss; overexpression; prefoldin; preservation; profilin 1; promoter; protein TDP-43; protein aggregation; rat genome; skeletal muscle wasting; sporadic amyotrophic lateral sclerosis; superoxide dismutase 1; transgene expression

Pathogenic mutation of profilin 1 (PFN1) is causative to amyotrophic lateral sclerosis (ALS). PFN1 has been examined extensively on its biology and recently on its pathobiology, but how PFN1 mutation causes the disease remains to be determined. A critical step towards understanding a disease gene is determining the overall effect of pathogenic mutation on the gene function at both systemic and molecular levels. ALS is a neurological disease and thus the overall effect of pathogenic mutation on PFN1 function must be determined in a vertebrate animal model because the complexity of the human central nervous system cannot be adequately simulated in cells or invertebrate animals. An effective animal model for ALS should meet at least three criteria: recapitulate the cardinal phenotypes of ALS including progressive degeneration of both upper and lower motor neurons and denervation atrophy of skeletal muscles; manifest the middle or late onset of ALS phenotypes that is common to sporadic and most inherited ALS; and display a differentiated disease phenotype between wild-type and mutant transgenic lines. An ideal animal model for human disease is gene knockin in which a single disease-causing mutation is introduced into the animal genome such that the pathogenic mutation is expressed at physiological levels and in intrinsic patterns, simulating the patterns and zygosities of gene expression that are observed in patients. As animals commonly used to model human diseases (i.e. mice and rats) have a shorter life span (2-3 years at maximum) than human (80 years on average), expression of a disease gene at physiological levels may not be sufficient to induce a full spectrum of disease phenotypes. Therefore, gene-overexpressing models (i.e. transgenics) are often used as the substitutes of knockin models for studying inherited diseases. To unravel the overall effect of pathogenic mutation on the PFN1 functions, we have created PFN1 transgenic rats that meet the criteria of effective ALS model and also have created PFN1 knockin rats that express PFN1 mutation from its endogenous locus. The knockin rats differ from their wildtype littermates in a single nucleotide examined. Any phenotypes detected in the knockin rats must result from the pathogenic mutation introduced. Using PFN1 transgenic and knockin rats as complementary models, we are going to determine the overall effect of pathogenic mutation on PFN1 function at both systemic and molecular levels, revealing the authentic mechanisms by which PFN1 mutation causes the disease. Our PFN1 rat models will be the second effective ALS model after SOD1 transgenic rodents and thus will meet the compelling need of ALS research in that disease mechanisms and therapeutic efficacies discovered in one model must be determined in the other models for the convergence and divergence among varying ALS genes because convergent disease mechanisms revealed in genetic ALS models will have a better prediction of sporadic ALS.

profilin 1（PFN1）的致病突变是肌萎缩性侧索硬化症（ALS）的原因。 pfn1曾经 对其生物学以及最近的病理生物学进行了广泛的研究，但是PFN1突变如何引起该疾病 仍有待确定。了解疾病基因的关键一步是确定总体效应 基因功能在全身和分子水平上的致病突变。 ALS是一种神经疾病 因此，必须在脊椎动物动物中确定致病突变对PFN1功能的总体影响 模型是因为人类中枢神经系统的复杂性无法在细胞或 无脊椎动物。 ALS的有效动物模型应符合至少三个标准：概括 ALS的基本表型，包括上和下运动神经元的进行性变性 骨骼肌的去神经萎缩；表现出ALS表型的中期或晚期发作 零星和大多数继承的ALS；并在野生型和突变体之间显示出分化的疾病表型 转基因线。人类疾病的理想动物模型是基因敲击素，其中单一引起疾病 突变被引入动物基因组中，使致病性突变在生理上表达 水平和内在模式，模拟基因表达的模式和合子性，在 患者。由于通常用于建模人类疾病的动物（即小鼠和大鼠）的寿命较短（2-3 比人（平均80年）最多的年度，生理水平的疾病基因的表达可能 不足以诱导各种疾病表型。因此，过表达基因的模型（即 转基因）通常用作研究遗传疾病的敲蛋白模型的替代品。解开 致病突变对PFN1功能的总体影响，我们创建了符合的PFN1转基因大鼠 有效ALS模型的标准，还创建了PFN1敲击大鼠，该大鼠从 它的内源基因座。在检查的单个核苷酸中，敲击大鼠与野生型同窝窝不同。任何 在敲击大鼠中检测到的表型必须是由于引入的致病性突变而引​​起的。使用PFN1 转基因和敲击大鼠作为互补模型，我们将确定致病性的总体作用 PFN1在全身和分子水平上功能的突变，揭示了真实的机制 PFN1突变引起该疾病。我们的PFN1大鼠模型将是SOD1之后的第二个有效ALS模型 转基因啮齿动物，因此将满足ALS研究在该疾病机制中的迫切需求，并且 在一个模型中发现的治疗效率必须在另一个模型中确定，以进行收敛和 不同的ALS基因之间的差异是因为遗传ALS揭示了收敛性疾病机制 模型将更好地预测零星的ALS。