Human Genetic Disease Modeling and Physiology Core

核心B -项目摘要 为了成功治疗α-1抗胰蛋白酶缺乏症，有必要使动物 在人类患者中看到的重现表型的模型，以测试治疗方法，包括 基因治疗和基因组编辑方法，并开发临床相关的治疗 以确定这些疗法的成功。为了实现这一目标，本核心正在与 项目1（Flotte），以创建条件性表达人AAT的AAT-peptide或PiZ雪貂 在本发明的实施方案中，使用了人AAT（hAAT）基因敲除的背景下的PiZ突变的鼠模型以及在AAT敲除的背景下的PiZ突变的鼠模型。 在我们组中创建的鼠标。 敲除小鼠重现了完全缺乏AAT产生的肺表型， 但是不能完全模拟具有一些残余血液的PiZ AAT患者的状态 产生残余抗弹性蛋白酶活性的AAT水平。在这个核心中，我们的目标是创造和 在AAT-KO背景下表征AAT-KO-PiZ小鼠，以及在AAT-KO背景下表征AAT-KO-PiZ小鼠。 现有的AAT-KO/PiZ交叉鼠标。除了作为一个更相关的模型来测试基因 AAT-KO-PiZ小鼠也将作为进一步研究基因组的模型。 编辑项目2中的Z-AAT等位基因（Xue）。当前的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。两种小鼠模型都允许 我们进一步阐明了PiZ突变在没有野生型小鼠基因的情况下所起的作用。 AAT，这是以前没有在小鼠中建模的东西。在核心B中，我们还将产生 人源化肝NSG-PiZ小鼠以选择靶向人肝细胞的AAV血清型， 项目3（王）。 除了生产用于项目1、2和3的小鼠和雪貂模型外，我们还将 我还与项目1和2一起工作，以表征肺表型对基因的反应， 治疗和基因组编辑。核心将提供新的专门知识，在啮齿动物肺功能 支气管肺泡灌洗试验和促进肺部计算机断层扫描， 与Bankier博士和Tufts Cummings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.