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.

项目概要 科学家可以利用越来越多的高通量制造出数千种化学性质不同的纳米粒子 化学，但在体内测试多个纳米颗粒仍然很困难。这项工作的目标是 通过执行系统性研究，显着改善脂质纳米颗粒 (LNP) 提供核酸治疗的方式 高通量体内 LNP 研究。这一目标将通过使用尖端的 DNA 条形码纳米颗粒来实现； 可以在一只小鼠中测量由 300 种不同纳米粒子介导的递送器。 4,320 化学性质不同 纳米颗粒将在体外和体内进行测试，重点关注两个基本问题。首先，如何 纳米颗粒结构影响体内细胞靶向？纳米粒子的化学和物理特性影响递送 体外。然而，相同的 LNP 特征在多大程度上影响动物（体内）递送尚不清楚。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 体内和体外巨噬细胞、内皮细胞和肝细胞的体内递送。相同的数据将 用于 (ii) 量化体外药物递送预测体内药物递送的精度。二、如何 临床相关的生理变化是否会影响体内递送？ LNP 与脂蛋白类似， 是天然的含脂质纳米结构。脂蛋白被主动运输至内皮细胞、巨噬细胞、 和体内肝细胞。鉴于高胆固醇患者服用他汀类药物后脂蛋白运输发生变化， 与患有许多其他病症的患者一样，LNP 的转运也可能发生变化。排名前 600 位的体内 LNP 4,320 LNP 体内筛选将应用于异常脂质运输的遗传小鼠模型，以便 (iii) 研究胆固醇运输的基因改变如何影响体内传递。这项工作将使5 对纳米技术做出了重大贡献。首先，LNP化学特性影响递送的程度 直接在体内进行测试；在体外研究了纳米粒子结构和递送之间的关系。 其次，体外纳米粒子递送预测体内递送的精确度将被量化。这 可以提高临床纳米颗粒的发现效率。三、临床效果 将检查 LNP 输送的相关生理变化。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，并可能影响纳米颗粒的靶向/ 安全。第四，将证明在体内研究数千个 LNP 的可行性。五、开源 将建立并传播纳米颗粒条形码协议。这些结果将提供至关重要的 深入了解 LNP 化学特性和特定基因改变 LNP 递送的方式，为 LNP 的设计提供信息 为多种治疗应用提供核酸货物（例如 siRNA、mRNA、CRISPR-Cas9）。

项目成果

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

用于将雾化治疗 mRNA 递送至肺部的脂质纳米粒子的优化。

• 发表时间: 2021-09
• 影响因子: 28.1
• 作者: Lokugamage, Melissa P;Vanover, Daryll;Beyersdorf, Jared;Hatit, Marine Z C;Rotolo, Laura;Echeverri, Elisa Schrader;Peck, Hannah E;Ni, Huanzhen;Yoon, Jeong;Kim, YongTae;Santangelo, Philip J;Dahlman, James E
• 通讯作者: Dahlman, James E

Therapeutic RNA Delivery for COVID and Other Diseases.

新冠肺炎和其他疾病的治疗性 RNA 递送。

• 发表时间: 2021-08
• 影响因子: 10
• 作者: Dobrowolski, Curtis;Paunovska, Kalina;Hatit, Marine Z C;Lokugamage, Melissa P;Dahlman, James E
• 通讯作者: Dahlman, James E

Nanoparticle stereochemistry-dependent endocytic processing improves in vivo mRNA delivery.

纳米颗粒立体化学依赖性内吞加工可改善体内 mRNA 递送。

• 发表时间: 2023-04
• 影响因子: 21.8
• 作者: Hatit, Marine Z C;Dobrowolski, Curtis N;Lokugamage, Melissa P;Loughrey, David;Ni, Huanzhen;Zurla, Chiara;Da Silva Sanchez, Alejandro J;Radmand, Afsane;Huayamares, Sebastian G;Zenhausern, Ryan;Paunovska, Kalina;Peck, Hannah E;Kim, Jinwhan;Sato
• 通讯作者: Sato

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。

• 发表时间: 2022-12-28
• 影响因子: 10.8
• 作者: Paunovska, Kalina;Da Silva Sanchez, Alejandro J;Lokugamage, Melissa P;Loughrey, David;Echeverri, Elisa Schrader;Cristian, Ana;Hatit, Marine Z C;Santangelo, Philip J;Zhao, Kun;Dahlman, James E
• 通讯作者: Dahlman, James E

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

体内对肺靶向脂质纳米颗粒的转录反应。

• 发表时间: 2023-02-08
• 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;Da Silva Sanchez, Alejandro J;Paunovska, Kalina;Schrader Echeverri, Elis
• 通讯作者: Schrader Echeverri, Elis