Understanding the Relationship LNP Structure, Cholesterol Trafficking, and InVivo Delivery

了解 LNP 结构、胆固醇运输和体内递送之间的关系

• 批准号: 10624289
• 负责人: James Dahlman
• 金额: $ 39.84万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10624289
• 关键词: Acrylates; Affect; Amines; Animals; Apolipoprotein E; Bar Codes; Biocompatible Materials; Bioinformatics; CRISPR/Cas technology; Cell Culture Techniques; Cell Line; Cells; Chemical Structure; Chemicals; Chemistry; Cholesterol; Clinical; Custom; DNA; DNA delivery; Data; Disease; Drug Delivery Systems; Dyslipidemias; Endothelial Cells; Epoxy Compounds; Feasibility Studies; Genes; Genetic; Genetic Models; Goals; Heart; Hepatocyte; High Fat Diet; Human; Immune response; In Vitro; Individual; Knockout Mice; Kupffer Cells; Lead; Length; Lipids; Lipoproteins; Liver; Lung; Macrophage; Measures; Mediating; Messenger RNA; Mus; Mutation; Nanostructures; Nanotechnology; Nucleic Acids; Organism; Pathway interactions; Patients; Physiological; Property; Protocols documentation; Safety; Scientist; Small Interfering RNA; Spleen; Structure; Testing; Therapeutic; Tissues; Toxic effect; Wild Type Mouse; Work; bioinformatics pipeline; cell type; cholesterol trafficking; clinical effect; clinically relevant; deep sequencing; design; experimental study; improved; in vitro testing; in vivo; insight; iterative design; lipid nanoparticle; lipid transport; mouse model; nanoparticle; nanoparticle delivery; nucleic acid-based therapeutics; open source; patient population; tertiary amine; trafficking; trait

Project Summary Scientists can create thousands of chemically distinct nanoparticles using a growing number of high throughput chemistries, but it is still difficult to test more than a few nanoparticles in vivo. The goal of this work is to substantially improve how lipid nanoparticles (LNPs) deliver nucleic acid therapies by performing a systematic high throughput in vivo LNP study. This goal will be achieved using cutting edge DNA barcoded nanoparticles; deliverer mediated by 300 different nanoparticles can be measured in a single mouse. 4,320 chemically distinct nanoparticles will be tested in vitro and in vivo, focusing on 2 fundamental questions. First, how does nanoparticle structure affect cell targeting in vivo? Nanoparticle chemical and physical traits affect delivery in vitro. However, the extent to which the same LNP traits influence delivery in animals (in vivo) is unclear. A recently developed bioinformatics pipeline will be used to (i) systematically analyze how LNP structure affects in in vivo delivery in macrophages, endothelial cells, and hepatocytes, both in vitro and in vivo. The same data will be used to (ii) quantify the precision with which in vitro drug delivery predicts in vivo drug delivery. Second, how do clinically relevant physiological changes affect delivery in vivo? LNPs are similar to lipoproteins, which are natural lipid-containing nanostructures. Lipoproteins are actively trafficked to endothelial cells, macrophages, and hepatocytes in vivo. Given that lipoprotein trafficking changes in patients with high cholesterol, taking statins, and patients with many other conditions, LNP transport may also change. The top 600 in vivo LNPs from the 4,320 LNP in vivo screen will be administered to genetic mouse models of aberrant lipid transport in order to (iii) investigate how genetic alterations in cholesterol trafficking affect in vivo delivery. This work will make 5 significant contributions to nanotechnology. First, the extent to which LNP chemical traits influence delivery directly in vivo will be tested; relationships between nanoparticle structure and delivery are studied in vitro. Second, the precision with which in vitro nanoparticle delivery predicts in vivo delivery will be quantified. This could increase the efficiency with which clinical nanoparticles are discovered. Third, the effect of clinically relevant physiological changes on LNP delivery will be examined. Nanoparticles can interact with cholesterol trafficking pathways; these interactions are likely to change with disease and can affect nanoparticle targeting / safety. Fourth, the feasibility of studying thousands of LNPs in vivo will be demonstrated. Fifth, open source protocols for nanoparticle barcoding will be established and disseminated. These results will provide crucial insight into the ways LNP chemical traits and specific genes alter LNP delivery, informing the design of LNPs that deliver nucleic acid cargos (e.g., siRNA, mRNA, CRISPR-Cas9) for numerous therapeutic applications.

项目摘要 科学家可以使用越来越多的高吞吐量来创建数千种化学上不同的纳米颗粒 化学，但是在体内测试多个纳米颗粒仍然很难。这项工作的目的是 实质上改善了脂质纳米颗粒（LNP）如何通过系统进行核酸疗法 高通量体内LNP研究。使用尖端DNA条形码纳米颗粒将实现此目标； 可以在单个小鼠中测量由300种不同纳米颗粒介导的递送子。 4,320化学不同 纳米颗粒将在体外和体内进行测试，重点关注2个基本问题。首先，如何 纳米颗粒结构会影响体内细胞靶向细胞的靶向？纳米颗粒化学和物理特征会影响递送 体外。但是，相同的LNP特征影响动物（体内）的递送的程度尚不清楚。一个 最近开发的生物信息学管道将用于（i）系统地分析LNP结构如何影响 体内体内的体内递送，内皮细胞和肝细胞，包括体外和体内。相同的数据将 用于（ii）量化体外药物递送在体内药物递送的精度。第二，怎么 临床上相关的生理变化会影响体内的分娩吗？ LNP与脂蛋白相似，脂蛋白 是天然含脂质的纳米结构。脂蛋白被积极运输到内皮细胞，巨噬细胞，巨噬细胞， 和体内的肝细胞。鉴于高胆固醇患者的脂蛋白运输变化，服用他汀类药物， 和有许多其他情况的患者，LNP运输也可能发生变化。来自 为了（iii） 研究胆固醇运输的遗传改变如何影响体内递送。这项工作将使5 对纳米技术的重要贡献。首先，LNP化学特征影响交付的程度 直接在体内进行测试；在体外研究了纳米颗粒结构与递送之间的关系。 其次，将量化体外纳米颗粒递送的精度将在体内递送。这 可以提高发现临床纳米颗粒的效率。第三，临床上的影响 将检查有关LNP递送的相关生理变化。纳米颗粒可以与胆固醇相互作用 贩运途径；这些相互作用可能会随着疾病而改变，并且可能影响纳米颗粒的靶向 / 安全。第四，将证明研究数千个LNP的可行性。第五，开源 将建立和传播纳米颗粒条形码的协议。这些结果将提供至关重要的 深入了解LNP化学性状和特定基因改变LNP的递送的方式，从而告知LNP的设计 为众多治疗应用提供核酸碳（例如siRNA，mRNA，CRISPR-CAS9）。

项目成果

The Transcriptional Response to Lung-Targeting Lipid Nanoparticles in Vivo.

• DOI: 10.1021/acs.nanolett.2c04479
• 发表时间: 2023-02-08
• 期刊: NANO LETTERS
• 影响因子: 10.8
• 作者: Radmand, Afsane;Lokugamage, Melissa P.;Kim, Hyejin;Dobrowolski, Curtis;Zenhausern, Ryan;Loughrey, David;Huayamares, Sebastian G.;Hatit, Marine Z. C.;Ni, Huanzhen;Del Cid, Ada;Sanchez, Alejandro J. Da Silva;Paunovska, Kalina;Echeverri, Elisa Schrader;Shajii, Aram;Peck, Hannah;Santangelo, Philip J.;Dahlman, James E.
• 通讯作者: Dahlman, James E.

Optimization of lipid nanoparticles for the delivery of nebulized therapeutic mRNA to the lungs.

• DOI: 10.1038/s41551-021-00786-x
• 发表时间: 2021-09
• 期刊: Nature biomedical engineering
• 影响因子: 28.1

Therapeutic RNA Delivery for COVID and Other Diseases.

• DOI: 10.1002/adhm.202002022
• 发表时间: 2021-08
• 期刊: Advanced healthcare materials
• 影响因子: 10
• 作者: Dobrowolski C;Paunovska K;Hatit MZC;Lokugamage MP;Dahlman JE
• 通讯作者: Dahlman JE

Substituting racemic ionizable lipids with stereopure ionizable lipids can increase mRNA delivery.

用立体纯可电离脂质替代外消旋可电离脂质可以增加 mRNA 递送。

• DOI: 10.1016/j.jconrel.2022.11.037
• 发表时间: 2023
• 期刊: Journal of controlled release : official journal of the Controlled Release Society
• 作者: DaSilvaSanchez,AlejandroJ;Zhao,Kun;Huayamares,SebastianG;Hatit,MarineZC;Lokugamage,MelissaP;Loughrey,David;Dobrowolski,Curtis;Wang,Shuaishuai;Kim,Hyejin;Paunovska,Kalina;Kuzminich,Yanina;Dahlman,JamesE
• 通讯作者: Dahlman,JamesE

The Extent to Which Lipid Nanoparticles Require Apolipoprotein E and Low-Density Lipoprotein Receptor for Delivery Changes with Ionizable Lipid Structure.

脂质纳米颗粒需要载脂蛋白 E 和低密度脂蛋白受体进行递送的程度随可电离脂质结构的变化而变化。

• DOI: 10.1021/acs.nanolett.2c03741
• 发表时间: 2022
• 期刊: Nano letters
• 影响因子: 10.8
• 作者: Paunovska,Kalina;DaSilvaSanchez,AlejandroJ;Lokugamage,MelissaP;Loughrey,David;Echeverri,ElisaSchrader;Cristian,Ana;Hatit,MarineZC;Santangelo,PhilipJ;Zhao,Kun;Dahlman,JamesE
• 通讯作者: Dahlman,JamesE