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：多(胺-共酯)S靶向输送基因编辑剂治疗动物囊性纤维化 模型：SCGE疾病模型研究补充资料 参与SCGE奖、研究人员和机构：UG3/UH3HL147352，POLY(胺-共酯)S 基因编辑剂体内定向输送到骨髓和肺，耶鲁大学，MPI：Wm Saltzman 和格雷泽首相。CO-I：我·伊根和R·范 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 开发了一种合成一族聚(胺-共酯)(PACE)聚合物的可靠方法，该聚合物可以 组装成纳米颗粒(NPs)，有效地包裹核酸。我们已经证明了这些NP是 将基因编辑剂运送到肺部的有效载体，以及编辑的效率-以及 经过编辑的特定肺细胞群-可以通过选择最佳的PACE组合进行优化 通过选择给药方式，静脉给药(IV)或气管内给药(IN)。在这里我们 将测试这些优化的传递系统以进行基因编辑，以治疗囊性纤维化(CF)。为此，我们有 在室内有两种不同的、特征良好的CF动物模型：一种包含最常见的囊性纤维化 跨膜电导调节基因突变(F508del)被归类为II类突变和 另一个包含一个常见的突变，它引入了一个被认为是I类突变的终止密码子(W1282X)。 这些动物的基因编辑将通过水滴数字聚合酶链式反应(DdPCR)和深度测序以及 直接测量肺和肠道的电生理，使用我们开发的方法。

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.