Modelling cerebellar pathology of Ataxia-Telangiectasia: Assessing ATM-deficient mice versus human iPS cells

共济失调-毛细血管扩张症(A-T)是由共济失调-毛细血管扩张症突变(ATM)基因突变引起的，表现为常染色体隐性遗传性多系统疾病，包括神经功能障碍。ATM被研究得最多的功能是DNA损伤反应的信号转导的主控制器。然而，ATM的额外功能可能在A-T的神经退行性表型中发挥关键作用，但迄今为止人们对此知之甚少。由于缺乏合适的体内(动物)和体外(细胞培养)模型来准确反映疾病的表型，许多疾病的分子基础，包括A-T，一直受到阻碍。重要的是，将体细胞重新编程为诱导多能干细胞(IPSCs)，然后在体外产生与疾病相关的细胞类型，为深入了解疾病的分子和细胞基础提供了机会。神经障碍尤其可以从IPSC疾病建模中受益匪浅，因为中枢神经系统的细胞类型，如神经元，通常只能从死后样本中获得。在这个项目中，两位申请者建立在他们互补的专业知识基础上(1)在小鼠大脑OMICS水平上分析影响浦肯野神经元功能的共济失调通路，(2)在IPSCs的高级基因工程方面。这个项目的目的是深入了解影响与A-T相关的神经退行性表型的细胞通路。从ATM-/-小鼠建立的器官类型切片培养将被用于研究DNA和氧化损伤诱导的细胞神经炎症和自噬变化，以及ATM的突触功能。此外，对ATM-/-和野生型小鼠小脑样本的转录组和蛋白质组分析将使我们能够以公正的方式发现和注释与ATM神经功能相关的新途径和标记。同时，将建立患者来源的IPSC系及其经过基因校正的对应物的等基因对，用于模拟A-T。这些同基因的细胞系将被用来衍生神经前体细胞，并在体外进行鉴定。该项目的成果将为疾病的通路机制提供有价值的见解，并为研究A-T相关的神经变性提供基于细胞的模型，这可能为未来的治疗提供新的靶点。

Ataxia-telangiectasia (A-T) is caused by mutations in the ataxia-telangiectasia mutated (ATM) gene, which manifest in an autosomal recessive multi-system disorder, including neurological dysfunction. The best-studied function of ATM is a master controller of signal transduction for DNA damage response. However, additional and thus far poorly understood functions of ATM likely play critical roles in the neurodegenerative phenotype of A-T. Understanding the molecular basis of many diseases, including A-T, has been hampered by the lack of appropriate in vivo (animal) and in vitro (cell culture) models that accurately reflect the disease phenotypes. Importantly, reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) followed by generation of disease-relevant cell types in vitro provides an opportunity to gain insight into the molecular and cellular basis of disease. Neurological disorders in particular can greatly benefit from iPSC-disease modeling, because cell types of the central nervous system, such as neurons, are typically available only from post-mortem samples.In this project the two applicants build upon their complementary expertise (1) in mouse brain OMICS-level analysis of ataxia pathways affecting Purkinje neuron function and (2) in advanced genetic engineering of iPSCs. The aim of this project is to gain detailed insight into the cellular pathways affecting the neurodegenerative phenotype associated with A-T.Organotypic slice cultures established from Atm-/- mice will be employed to address DNA- and oxidative damage-induced cellular neuroinflammation and autophagy changes, as well as synaptic functions of ATM. Furthermore, transcriptome and proteome analyses of cerebellar samples of Atm-/- and wild-type mice will enable us to discover and annotate novel pathways and markers associated with neuronal functions of ATM in an unbiased manner. In parallel, isogenic pairs of patient-derived iPSC lines and their genetically corrected counterparts will be established for modeling A-T. The isogenic pairs of cell lines will be used to derive neuronal progenitors and characterized in vitro. The output of this project will provide valuable insights into the pathway mechanisms of disease, and cell-based models to investigate A-T-associated neurodegeneration, which may provide novel targets for future therapies.