Poly(amine-co-ester)s for targeted delivery of gene editing agents to treat cystic fibrosis in animal models: SCGE Disease Models Studies Supplement

项目名称：用于靶向递送基因编辑剂以治疗动物囊性纤维化的聚（胺-共-酯） 模型：SCGE疾病模型研究补充 参与SCGE奖项的研究者和机构：UG3/UH3 HL 147352，聚（胺-共-酯），用于 基因编辑剂体内靶向递送到骨髓和肺，耶鲁大学，MPI：WM Saltzman 和格雷泽首相Co-I：ME埃根和R Fan 翻译后摘要：在过去的几年里，在NIH体细胞基因编辑联盟的支持下，我们已经 开发了用于合成家族聚（胺-共-酯）（PACE）聚合物的稳健方法， 组装成纳米颗粒（NP），其有效地捕获核酸。我们已经证明，这些NP是 用于将基因编辑剂递送到肺的有效载体，以及编辑的效率以及 编辑的特定肺细胞群可以通过选择PACE的最佳组合来优化 变体，以及通过选择静脉内（IV）或腹膜内（IN）给药模式。这里我们 将测试这些优化的基因编辑递送系统，以治疗囊性纤维化（CF）。为此，我们 在室内两种不同的，良好表征的CF动物模型：一种包含最常见的囊性纤维化 跨膜传导调节因子（CFTR）基因突变（F508del），分类为II类突变， 另一个包含引入终止密码子（W1282X）的常见突变，其被认为是I类突变。 这些动物中的基因编辑将通过液滴数字PCR（ddPCR）和深度测序来检测，以及 使用我们开发的方法直接测量肺和肠道的电生理学。

Project title: Poly(amine-co-ester)s for targeted delivery of gene editing agents to treat cystic fibrosis in animal models: SCGE Disease Models Studies Supplement Participating SCGE award, investigators, and institutions: UG3/UH3 HL147352, Poly(amine-co-ester)s for targeted delivery in vivo of gene editing agents to bone marrow and lung, Yale University, MPI: WM Saltzman and PM Glazer. Co-I: ME Egan and R Fan Abstract: Over the past years, with support from the NIH Somatic Cell Gene Editing consortium, we have developed robust methods for synthesis of a family poly(amine-co-ester) (PACE) polymers that can be assembled into nanoparticles (NPs) that efficiently entrap nucleic acids. We have shown that these NPs are effective vectors for delivery of gene editing agents to the lung, and that the efficiency of editing—as well as the specific lung cell populations that are edited—can be optimized by the selection of optimal combinations of PACE variants, and by selection of the mode of administration, either intravenous (IV) or intratracheal (IN). Here we will test these optimized delivery systems for gene editing to treat cystic fibrosis (CF). For that purpose, we have in house two different, well-characterized animal models of CF: one contains the most common cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation (F508del) classified as a Class II mutation and the other contains a common mutation that introduces a stop codon (W1282X) considered a Class I mutation. Gene editing in these animals will be detected by droplet digital PCR (ddPCR) and deep sequencing, as well as direct measurement of electrophysiology of the lung and the gut, using approaches we have developed.