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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

基本信息

批准号：
10172933
负责人：
James Dahlman
金额：
$ 37.18万
依托单位：
GEORGIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
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/10172933
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 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 macrophage mouse model nanoparticle nanoparticle delivery open source patient population tertiary amine trafficking trait

项目摘要

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.

项目总结