Efficient in Vivo RNP-based Gene Editing in the Sensory Organ Inner Ear Using Bioreducible Lipid Nanoparticles

使用生物可还原脂质纳米颗粒对感觉器官内耳进行基于 RNP 的高效体内基因编辑

基本信息

批准号：
10470326
负责人：
Zheng-Yi Chen
金额：
$ 163.97万
依托单位：
MASSACHUSETTS EYE AND EAR INFIRMARY
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：
2018-09-18 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10470326
关键词：
Address Affect Animal Model Animals Biological Biological Assay Cations Cells Characteristics Clinic Clinical Defect Detection Disease Environment Epithelial Evaluation Event Family suidae Formulation Freeze Drying Gene Delivery Gene Mutation Genes Hair Cells Hearing Human In Vitro Individual Investigation Labyrinth Libraries Link Lipids Liposomes Mediating Methods Microfluidics Modality Modeling Mus Neurons Newborn Infant Nonhomologous DNA End Joining Organ Organ Model Phase Process Production Proteins Resolution Ribonucleoproteins Sensory Sensory Hair Somatic Cell Specificity Structure System Technology Testing Tissues Toxic effect Translations Work base base editing biomaterial compatibility cell type clinical application clinical development combinatorial design disulfide bond effective therapy genome editing hereditary hearing loss human disease human model human tissue improved in vivo innovation large scale production lipid nanoparticle male mouse model nanoparticle nanoparticle delivery novel porcine model prevent screening therapeutic genome editing therapy development

项目摘要

Project Summary and Relevance Application of genome editing based therapy requires efficient delivery of editing agents into disease-relevant tissues and cells. Identification of novel delivery materials targeting somatic cells will greatly facilitate the advance of editing based therapy strategies to clinic. We propose to screen a large library of novel lipid nanoparticles for RNP (ribonucleoprotein) delivery of editing agents into the mammalian sensory organ inner ear. Inner ear is an ideal sensory organ to develop new delivery strategies. It consists of multiple differentiated somatic cell types without effective delivery options. Gene mutations in the major inner ear cell types have been associated with genetic hearing loss, which affects one in 500 newborns and currently has no effective therapies. Lipid-based nanoparticle carriers have emerged as one of the most promising materials for delivery and have been successfully used in clinical applications. We have developed a combinatorial library approach to synthesize degradable lipid-like nanoparticles under reductive intracellular environments, and capable of delivering biomolecules with high efficiency and low toxicity. The new bioreducible lipid nanoparticles (bLNPs) have been used to deliver genome editing agents with high efficiency and low toxicity in vivo. We have delivered genome editing RNP by cationic liposomes into mammalian inner ear in vivo, and rescued hearing in mouse models of human genetic hearing loss. To develop editing based therapies to treat diverse forms of genetic hearing loss, it is essential to develop a delivery strategy to target multiple inner ear cell types simultaneously. The mammalian inner has a complicated structure with multiple cell types in small numbers, making it particularly challenging to screen a delivery technology by conventional high-throughput strategies. The lack of a method to detect editing at the level of the individual cell type further hinders our ability to apply this technology in wildtype large animal models that are essential for development of this therapy for clinical application. By combining our strategies to screen nanoparticles for delivery of editing materials to X-linked genes in the male mouse inner ears in vivo, we will overcome these hurdles for effective delivery and editing in diverse inner ear cell types. The study of the human inner ear tissues ex vivo will provide evidence of the relevance of nanoparticle delivery in human disease-relevant tissues. Expansion of our work to large animal models will be a major step towards clinical application of this technology. Our approach with nanoparticles can be applied to the study in other organs requiring somatic cell type editing and in wildtype large animals.

项目摘要和相关性基于基因组编辑的治疗的应用需要有效地将编辑剂递送到疾病相关的组织和细胞。鉴定针对体细胞的新型递送材料将极大地促进基于编辑的治疗策略提前到诊所。我们建议筛选一个大型新型脂质库 RNP的纳米颗粒（核糖核蛋白）递送编辑剂到哺乳动物感觉器官内部耳朵。内耳是制定新交付策略的理想感官器官。它由多个差异化组成体细胞类型没有有效的交付选项。主要内耳细胞类型中的基因突变已经与遗传听力损失相关，这影响了500名新生儿，目前没有有效疗法。基于脂质的纳米颗粒载体已成为交付最有前途的材料之一，并具有成功用于临床应用。我们已经开发了一种组合库的方法在还原性细胞内环境下合成可降解的脂质的纳米颗粒，并且能够提供高效率和低毒性的生物分子。新的生物治疗脂质纳米颗粒（BLNP）已用于在体内提供高效率和低毒性的基因组编辑剂。我们已通过阳离子脂质体将基因组编辑RNP输送到体内的哺乳动物内耳中，并被救出在人类遗传听力损失的小鼠模型中听力。开发基于编辑的疗法以治疗多样化遗传听力损失的形式，必须制定递送策略以靶向多种内耳细胞类型同时地。哺乳动物内部具有复杂的结构，有多种细胞类型的数量很少，通过传统的高通量策略筛选交付技术的筛查特别具有挑战性。缺乏检测在单个单元类型级别的编辑的方法进一步阻碍了我们申请的能力野生型大型动物模型中的这项技术对于开发临床疗法至关重要应用。通过结合我们筛选纳米颗粒以将编辑材料交付到X连锁的策略雄性小鼠内耳中的基因在体内，我们将克服这些障碍，以有效分娩和编辑多种内耳细胞类型。人体内耳组织的研究将提供证据纳米粒子在与人类疾病相关的组织中的相关性。将我们的工作扩展到大动物模型将是该技术临床应用的主要一步。我们使用纳米颗粒的方法可以在需要躯体细胞类型编辑和野生型大动物的其他器官中应用于研究。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Clinical progress in genome-editing technology and in vivo delivery techniques.

DOI：
10.1016/j.tig.2022.12.001
发表时间：
2023-01
期刊：
Trends in genetics : TIG
影响因子：
0
作者：
Jennifer Khirallah;Maximilan Eimbinder;Yamin Li;Qiaobing Xu
通讯作者：
Jennifer Khirallah;Maximilan Eimbinder;Yamin Li;Qiaobing Xu

Lung-selective mRNA delivery of synthetic lipid nanoparticles for the treatment of pulmonary lymphangioleiomyomatosis.

DOI：
10.1073/pnas.2116271119
发表时间：
2022-02-22
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Qiu M;Tang Y;Chen J;Muriph R;Ye Z;Huang C;Evans J;Henske EP;Xu Q
通讯作者：
Xu Q

Protein and mRNA Delivery Enabled by Cholesteryl-Based Biodegradable Lipidoid Nanoparticles.