Poly(amine-co-ester)s for Targeted Delivery In Vivo of Gene Editing Agents to Bone Marrow and Lung

用于将基因编辑剂体内靶向递送至骨髓和肺的聚(胺-共酯)

基本信息

批准号：
10706300
负责人：
PETER M GLAZER
金额：
$ 114.48万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-07 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10706300
关键词：
Amines Animal Model Animals Binding Biocompatible Materials Biodistribution Biology Biomedical Engineering Bone Marrow CRISPR/Cas technology Cells Characteristics Charge Clinic Clinical Trials Collaborations Cystic Fibrosis DNA DNA Repair DNA Sequence Alteration Development Disease Disease model Encapsulated Esters Exhibits Family Family suidae Formulation Frequencies Gene Delivery Genes Genetic Diseases Genome Genome engineering Genomic DNA Genomics Glycolates Goals Guide RNA Health Hematopoietic stem cells Hereditary Disease Human In Vitro Inherited Injections Interdisciplinary Study Intravenous Laboratories Libraries Lung Measures Mediating Messenger RNA Methods MicroRNAs Modification Mus Nanotechnology Nucleic Acids Nucleotide Excision Repair Oligonucleotides Pathway interactions Patients Peptide Nucleic Acids Phase Polymers Process Protocols documentation Public Health Reagent Research Research Personnel Safety Site Small Interfering RNA Somatic Cell Structure Surface Symptoms System Technology Testing Therapeutic Tissues Toxic effect Translations United States National Institutes of Health Validation Work base base editing beta Thalassemia burden of illness cell type clinical translation deep sequencing delivery vehicle design disease-causing mutation engineered nucleases genome editing homologous recombination improved in vivo innovation intravenous injection nanoparticle nanosized new technology nonhuman primate novel nuclease plasmid DNA programs repaired response scale up screening single-cell RNA sequencing success targeted delivery therapeutic gene therapeutic genome editing

项目摘要

Project Summary Hereditary disorders such as β-thalassemia and cystic fibrosis are attractive targets for genome engineering as these maladies are curable upon correction of the disease-causing mutation. New technologies can catalyze correction at the associated genomic site by homologous recombination (HR); for example, engineered nucleases including CRISPR/Cas9 systems have shown promise and entered clinical trials. Alternative non- nuclease-based triplex-forming peptide nucleic acids (PNAs) have also been successful in vivo. PNAs have no intrinsic nuclease activity and enable activation of endogenous DNA repair activity when bound adjacent to the target site and co-delivered with a donor DNA strand containing the corrected sequence. PNA-mediated gene editing occurs via nucleotide excision repair (NER) and HR pathways and exhibits low off-target effects. While these editing technologies have been successful thus far, important challenges remain before translation to the clinic. The development of safe and effective delivery vehicles that are able to efficiently encapsulate gene editing agents and target disease-relevant cells/tissues is necessary for the advancement of these therapeutics. The goal of this research is to further the translation of genome engineering technologies by developing biodegradable poly(amine-co-ester) (PACE) into polymeric vehicles that efficiently encapsulate and deliver gene editing agents to target cells in the bone marrow and the lung upon systemic intravenous (IV) administration. PACE is structurally diverse, allowing us to generate libraries of vehicles and identify compositions for targeting bone marrow or lung. In preliminary work, we have observed efficient encapsulation and delivery of gene editing agents using PACE. Further, specific PACE formulations have exhibited favorable biodistribution to the bone marrow and lung. The project will proceed in two phases: a development phase (UG3) and a demonstration phase (UH3). In the UG3 phase, a library of PACE polymers with unique characteristics will be synthesized and tested for their ability to encapsulate PNA- and CRISPR/Cas9-based editing reagents, deliver them to target cells, and promote efficient editing in vitro and in vivo. Cell-type targeting and editing will be quantified using an innovative high-throughput single cell RNA sequencing (scRNA-seq) screen. Candidate formulations targeting bone marrow and lung will be administered to murine disease models of β-thalassemia and cystic fibrosis, respectively, to confirm their editing capabilities and determine their ability to ameliorate disease symptoms. In the UH3 phase, candidate formulations and gene editing agents will be scaled up to accommodate large animal studies, primarily in pigs, but also in non- human primates in collaboration with other investigators in the SCGE program. These studies are designed to confirm cell-type targeting, using quantitative measures of gene editing, and disease improvement, enabling key steps towards clinical trials. This interdisciplinary research will yield a platform of targeted delivery vehicles, furthering the translation of gene editing therapeutics for diseases resulting from genetic mutations.

项目概要 β-地中海贫血和囊性纤维化等遗传性疾病是基因组工程的有吸引力的目标这些疾病在纠正致病突变后是可以治愈的，新技术可以起到催化作用。例如，通过同源重组（HR）在相关基因组位点进行校正；包括 CRISPR/Cas9 系统在内的核酸酶已显示出前景并已进入临床试验。基于核酸酶的三链体形成肽核酸（PNA）在体内也取得了成功。内在核酸酶活性，并在与邻近的核酸酶结合时能够激活内源性 DNA 修复活性靶位点并与含有 PNA 介导的基因的供体 DNA 链共同递送。编辑通过核苷酸切除修复 (NER) 和 HR 途径进行，并且表现出较低的脱靶效应。到目前为止，这些编辑技术已经取得了成功，但在转化为现实之前仍然存在重要挑战开发能够有效封装基因的安全有效的递送载体。编辑剂和目标疾病相关细胞/组织对于这些疾病的进展是必要的这项研究的目标是进一步转化基因组工程技术。通过将可生物降解的聚（胺-酯）（PACE）开发成聚合物载体，可以有效地封装基因编辑剂并将其递送至骨髓和肺中的靶细胞全身静脉 (IV) 给药在结构上是多样化的，使我们能够生成库。在初步工作中，我们观察到了载体并确定了靶向骨髓或肺的组合物。使用 PACE 有效封装和递送基因编辑剂此外，还采用特定的 PACE 配方。对骨髓和肺部有良好的生物分布该项目将分两个阶段进行：开发阶段（UG3）和示范阶段（UH3）。将合成具有独特特性的材料并测试其封装 PNA- 和基于CRISPR/Cas9的编辑试剂，将其递送至靶细胞，促进体外和体内的高效编辑体内细胞类型靶向和编辑将使用创新的高通量单细胞 RNA 进行量化。将进行针对骨髓和肺的候选制剂的测序（scRNA-seq）筛选。分别对β-地中海贫血和囊性纤维化的小鼠疾病模型进行研究，以确认它们的编辑能力并确定其在 UH3 阶段改善疾病症状的能力。基因编辑剂将扩大规模以适应大型动物研究，主要是猪，但也适用于非这些研究旨在与 SCGE 项目的其他研究人员合作进行人类灵长类动物的研究。使用基因编辑和疾病改善的定量测量来确认细胞类型靶向，从而使这项跨学科研究将产生一个有针对性的交付平台。载体，进一步促进基因编辑疗法治疗基因突变引起的疾病的转化。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（1）

Compartmentalized ocular lymphatic system mediates eye-brain immunity.

区室化的眼淋巴系统介导眼脑免疫。

DOI：
发表时间：
2024-04
期刊：
影响因子：
64.8
作者：
Yin, Xiangyun;Zhang, Sophia;Lee, Ju Hyun;Dong, Huiping;Mourgkos, George;Terwilliger, Gordon;Kraus, Aurora;Geraldo, Luiz Henrique;Poulet, Mathilde;Fischer, Suzanne;Zhou, Ting;Mohammed, Farrah Shalima;Zhou, Jiangbing;Wang, Yongfu;Malloy, Seth
通讯作者：
Malloy, Seth