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Principal Investigator: James Dahlman
Organization: GEORGIA INSTITUTE OF TECHNOLOGY
Fiscal Year: 2020
Award: $370,620
Funding agency: National Institute of General Medical Sciences
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.
Terms: <Acrylates><Affect><Amines><Animals><Apo-E><ApoE><Apolipoprotein E><Bar Codes><Bio-Informatics><Biocompatible Materials><Bioinformatics><Biomaterials><Body Tissues><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Cell Body><Cell Culture Techniques><Cell Line><CellLine><Cells><Chemical Structure><Chemicals><Chemistry><Cholesterol><Clinical><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Custom><DNA><DNA delivery><Data><Deoxyribonucleic Acid><Disease><Disorder><Drug Delivery><Drug Delivery Systems><Dyslipidemias><Endothelial Cells><Epoxides><Epoxy Compounds><Feasibility Studies><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic Models><Genetic defect><Goals><Heart><Hepatic Cells><Hepatic Parenchymal Cell><Hepatocyte><High Fat Diet><Human><Immune response><Immunological response><In Vitro><Individual><KO mice><Knock-out Mice><Knockout Mice><Kupffer Cells><Lead><Length><Lipid Trafficking><Lipids><Lipoproteins><Liver><Liver Cells><Lung><Lung Respiratory System><Measures><Mediating><Messenger RNA><Mice><Mice Mammals><Modern Man><Murine><Mus><Mutation><Nanostructures><Nanotechnology><Nucleic Acids><Null Mouse><Organism><Pathway interactions><Patients><Pb element><Physiologic><Physiological><Property><Protocol><Protocols documentation><Safety><Scientist><Short interfering RNA><Small Interfering RNA><Spleen><Spleen Reticuloendothelial System><Stellate Sinusoidal Macrophage><Strains Cell Lines><Structure><Testing><Therapeutic><Tissues><Toxic effect><Toxicities><Wild Type Mouse><Work><amine><barcode><bio-informatics pipeline><bioinformatics pipeline><biological material><cell culture><cell type><cholesterol trafficking><clinical effect><clinical relevance><clinically relevant><cultured cell line><deep sequencing><deliver DNA><design><designing><experiment><experimental research><experimental study><genome mutation><heavy metal Pb><heavy metal lead><hepatic body system><hepatic organ system><host response><immunoresponse><improved><in vitro testing><in vivo><insight><iterative design><lipid nanoparticle><lipid transport><liver macrophage><living system><mRNA><macrophage><mouse model><murine model><nano particle><nano particle delivery><nano tech><nano technology><nano-sized particle><nano-structures><nano-technological><nanoparticle><nanoparticle delivered><nanoparticle delivery><nanosized particle><nanotech><nanotechnological><open source><pathway><patient population><pulmonary><siRNA><tertiary amine><trafficking><trait><wildtype mouse>