SCGE Disease Models Studies Supplement: Evaluation of prime editing for the amelioration of alpha-1-antitrypsin deficiency in murine and porcine models.

参与奖项、赞助商和机构 4UH 3 TR 002668 -04，“通过化学方法增强体细胞组织中的CRISPR基因编辑” 修改指南和捐助者”，埃里克Sontheimer博士和斯科特沃尔夫，大学 马萨诸塞州陈医学院 1U 24 OD 026641 -01，“用于检测体细胞基因组的猪报告基因模型的开发 编辑工具。"，丹尼尔卡尔森和杰瑞德坎贝尔博士，医学 摘要 α-1抗胰蛋白酶缺乏症（A1 AT）是一种遗传性疾病， SERPINA 1（丝氨酸蛋白酶抑制剂A1），其特征是低血清和低肺 AAT水平。最常见的突变是“Z”（Glu 342 Lys）等位基因，其在哺乳动物中降低AAT。 由于肝脏中的滞留导致肺中蛋白酶过度活跃而引起的循环，以及 突变错误折叠的AAT蛋白在肝细胞中的积累。这导致组织破坏 在许多A1 AT患者中，肺（功能丧失）和肝脏疾病（毒性功能获得）。 超过95%的严重A1 AT患者是Z等位基因纯合子（PI*ZZ）。 理想的治疗方法是修复PiZ等位基因。不幸的是，A1 AT基因治疗，即使 一种靶病变（肺或肝）尚未有效转化为患者， 在A1 AT的早期小鼠模型中有希望的数据。我们的团队汇集了A1 AT基因的专业知识 治疗、临床前动物模型、RNA递送和引物编辑技术。我们建议 开发优化的引物编辑试剂以修复PiZ等位基因。有了这些试剂， 在两种独特的A1 AT小鼠模型中进行剂量递增研究，以测量基因修复 结果以及对肺和肝表型的后续影响。小鼠的最佳剂量 将在我们的PiZ等位基因的新型人源化猪模型中进行评估，主要目标是 评估安全性和目标锁定情况我们相信这种逐步的治疗模式 在两种动物模型中的评价将提高在随后的研究中成功结果的可能性。 临床研究。

PARTICIPATING AWARDS, INVESTIGATORS & INSTITUTIONS 4UH3TR002668-04, “Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors”, Drs. Erik Sontheimer and Scot Wolfe, University of Massachusetts Chan Medical School 1U24OD026641-01, “Development of Swine Reporter Models for Testing Somatic Cell Genome Editing Tools.”, Drs. Daniel Carlson and Jarryd Campbell, Recombinetics ABSTRACT Alpha-1 antitrypsin deficiency (A1AT) is a hereditary disorder caused by point mutations in SERPINA1 (SERine Proteinase INhibitor A1) that is characterized by low serum and low lung levels of AAT. The most common mutation is the “Z” (Glu342Lys) allele that reduces AAT in circulation due to retention in the liver leading to overactive proteases in the lung, and accumulation of the mutant misfolded AAT protein in hepatocytes. This leads to tissue destruction in the lungs (loss-of-function), and liver disease (gain-of-toxic-function) in many A1AT patients. More than 95% of people with severe A1AT are homozygous for the Z allele (PI*ZZ). The ideal therapeutic would repair the PiZ allele. Unfortunately, A1AT gene therapy to treat even one target pathology (lung or liver) has not yet been effectively translated to patients despite promising data in early mouse models of A1AT. Our team brings together expertise in A1AT gene therapy, preclinical animal models, RNA delivery, and prime editing technology. We propose to develop optimized prime editing reagents to repair the PiZ allele. With these reagents, we will perform dosage escalation studies in two unique mouse models of A1AT to measure gene repair outcomes and the subsequent effect on lung and liver phenotypes. The optimal dose from mice will be evaluated in our novel, humanized swine model of the PiZ allele with the primary goal of evaluating safety and target engagement. We believe this stepwise paradigm for therapeutic evaluation in two animal models will improve the likelihood of a successful outcome in subsequent clinical studies.