Cell Reports: Nanoparticle-assisted CRISPR gene editing of vascular endothelium for the first time

Vascular endothelium is a single layer of endothelial cells lining the luminal surface of blood vessels, and the vascular endothelium plays a crucial role in vascular homeostasis and interstitial fluid homeostasis
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Endothelial dysfunction is at the root of many diseases, such as coronary artery disease, stroke, bronchopulmonary dysplasia and pulmonary hypertension, etc.

CRISPR-Cas9 is a powerful genome editing tool that has achieved impressive results in both basic research and clinical applications
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Adeno-associated virus (AAV) is the most important delivery vector for gene editing in vivo, but AAV also has some problems, such as inducing an immune response and making it difficult for secondary administration, and it is difficult to deliver gene sequences exceeding 4.

To date, efficient gene editing of the vascular endothelium has been difficult due to the lack of delivery systems targeting the vascular endothelium beyond the liver and eye
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On January 4, 2022, You-Yang Zhao's team at Northwestern University Feinberg School of Medicine published a paper entitled: Robust genome editing in adult vascular endothelium by nanoparticle delivery of CRISPR-Cas9 plasmid DNA in Cell Reports, a sub-journal of Cell.
research paper
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The research team developed a new type of nanoparticle, PEG-b-PLGA, to systematically deliver CRISPR-Cas9 gene editing to vascular endothelial cells, and successfully performed gene editing, which is the first time to edit vascular endothelial cells in vivo
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The research team developed PEG-b-PLGA nanoparticles configured from polyethyleneimine (PEI)
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The nanoparticles have excellent biodistribution for vascular delivery without specific liver aggregation

The team injected the nanoparticles, carrying the CRISPR-Cas9 gene-editing plasmid, into adult mice intravenously
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The delivery system induced efficient gene editing in endothelial cells of adult mouse vasculature, including lung, heart, aorta, and peripheral blood vessels, and the results showed that the protein expressed by the PI3KCG gene in the edited vascular endothelial cells decreased About 80%, and the resulting phenotype is similar to knockout mice

In addition, the gene editing plasmid delivered by the nanoparticle delivery system can induce the editing of at least two genes at the same time, and can also introduce foreign genes into vascular endothelial cells
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Professor Zhao, who led the study, said that the nanoparticle developed by the team is a powerful new vascular endothelial cell genome editing delivery system that can be used to treat a variety of diseases, including COVID-19
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Through this nanoparticle delivery system, exogenous genes can be introduced to inhibit vascular damage and/or promote vascular repair, and gene editing systems can be delivered to correct gene mutations and turn specific genes on or off

Professor Zhao said that the nanoparticles developed in this study can rapidly change gene expression in vascular endothelial cells by delivering plasmids
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It is an important breakthrough in the treatment of diseases caused by endothelial dysfunction and an important breakthrough in the field of cardiovascular research

Xianming Zhang, et al.