Human Genetic Disease Modeling and Physiology Core

核心B--项目摘要 为了有成功的治疗α-1抗胰蛋白酶缺乏症的方法，必须有动物 重述在人类患者身上看到的表型以测试治疗方法的模型，包括 基因治疗和基因组编辑方法，并开发临床相关的治疗方法 确定这些疗法成功与否的基准。为了达到这一目标，本核心正在与 项目1(Flotte)创建有条件表达人类AAT的AAT-Null或Piz雪貂 (Haat)以及以AAT基因敲除为背景的Piz突变的小鼠模型 在我们组中创建的老鼠。 基因敲除的小鼠重述了完全缺乏AAT产生的肺表型， 但不能完全模拟PIZ AAT患者的状态，这些患者有一些残余血液 产生残余抗弹力酶活性的AAT水平。在这个核心中，我们的目标是创建和 在AAT-KO背景上对AAT-KO-PIZ小鼠进行表征以及对AAT-KO-PIZ小鼠进行 现有的AAT-KO/PIZ杂交鼠标。除了作为一个更相关的模型来测试基因 治疗方面，AAT-KO-Piz小鼠也将作为进一步研究基因组的模型 项目2(XUE)中Z-AAT等位基因的编辑。当前Piz鼠标包含多个副本 PIZ基因，使其成为基因组编辑的非理想模型。这两种鼠标型号都将允许 美国将进一步阐明在没有野生型小鼠存在的情况下PIZ突变所起的作用 AAT，以前没有在鼠标上模拟过的东西。在酷睿B中，我们还将生产 人源化肝脏NSG-PIZ小鼠用于选择以人肝细胞为靶点的AAV血清型 项目3(王)。 除了生产用于项目1、2和3的老鼠和雪貂模型外，我们还将 我还与项目1和项目2一起工作，以表征肺对基因的表型反应 治疗和基因组编辑。核心将提供啮齿动物肺功能方面的新专业知识 肺CT检查、支气管肺泡灌洗试验及便利化 与班克尔博士和塔夫茨·卡明斯兽医学院合作。

Core B - Project Summary To have successful therapies for Alpha-1 antitrypsin deficiency it is necessary to have animal models that recapitulate the phenotypes see in human patients to test therapeutics, including gene therapy and genome editing approaches, and develop clinically relevant therapeutic benchmarks to ascertain the success of those therapies. To that aim this Core is working with Project 1 (Flotte) to create an AAT-Null or PiZ ferret that conditionally expresses human AAT (hAAT) as well as a murine model of the PiZ mutation on the background of the AAT knock-out mouse created in our group. The knock-out mouse recapitulates a lung phenotype with its complete lack of AAT production, but does not completely model the state of the PiZ AAT patient who have some residual blood levels of AAT that produce residual anti-elastase activity. In this Core we aim to create and characterize an AAT-KO-PiZ mouse on the AAT-KO background and well as characterize an existing AAT-KO/PiZ cross mouse. In addition to being a more relevant model to test gene therapies, the AAT-KO-PiZ mouse will also serve as the model to further investigate genome editing of the Z-AAT alleles in Project 2 (Xue). The current PiZ mouse contains multiple copies of the PiZ gene, making it a non-ideal model for genome editing. Both mouse models will allow us to elucidate further the role that the PiZ mutation plays without the presence of wild-type murine AAT, something that has not been modeled in the mouse before. In Core B we will also produce humanized liver NSG-PiZ mice to select for AAV serotypes that target human hepatocytes for Project 3 (Wang). In addition to the production of mouse and ferret models for use in Projects 1, 2, and 3 we will also work with Projects 1 and 2 to characterize the pulmonary phenotype responses to gene therapy and genome editing. The core will offer novel expertise in rodent pulmonary function testing, bronchoalveolar lavage testing and facilitation of lung computed tomography in collaboration with Dr. Bankier and Tufts Cummings School of Veterinary Medicine.