# Endosomal escape of lipid-based nanoparticles comprising Gaussian curvature lipids

> **NIH NIH R01** · UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN · 2023 · $167,855

## Abstract

PROJECT SUMMARY
We propose the acquisition of a new image-enhanced flow cytometer (Attune CytPix) to support the research of
1R01GM143723-01A1: Endosomal escape of lipid-based nanoparticles comprising Gaussian curvature lipids.
RNA-based therapies offer significant potential for treating a variety of diseases that have a major impact on
human health. These include chronic infections, genetic disorders, specific cancers, and the current COVID-19
pandemic. Non-viral lipid-based nanoparticles (LNPs) are the primary RNA delivery vehicles approved by the
FDA and are also being evaluated in numerous clinical trials. LNPs are composed of standard phospholipids,
cholesterol, and ionizable lipids (ILs) that become protonated in acidic conditions. Similar to enveloped viruses,
LNPs exploit the endocytic pathway to gain entry into cells. The success of RNA delivery depends on the ability
of LNPs to fuse with the endosomal membrane and escape the endosome. However, the mechanisms that
govern LNP-endosome fusion remain largely unknown.
The central goal of 1R01GM143723-01A1 is to test the hypothesis that the inclusion of a new class of structural
lipids with single chains and small headgroups like glycerol monooleate onto state-of-the art LNP formulations
allows us to prescribe well-defined internal nanostructures of LNPs directly impacting their ability to fuse with
endosomal membranes and releasing RNA cargo into the cytosol.
Acquiring the new Attune CytPix equipment is vital to the progression of our project for two main reasons. Firstly,
the only flow cytometry instrument in close proximity to our primary research location is currently non-operational
due to frequent breakdowns and high user demand, resulting in prolonged periods of downtime. Secondly, the
Attune CytPix offers simultaneous high throughput flow cytometry and high resolution brightfield imaging through
acoustic focusing, which is not available with our current equipment. This feature is particularly important for
quantifying LNP cargo loading and delivery efficiency. The high-speed brightfield camera records individual
events as they pass through the flow cell, and the Attune Cytometric Software ensures that analyzed events
originate from single cells and particles rather than doublets, clumps, or debris. This capability is critical in cell
and gene therapy research, as well as other flow cytometry experiments that aim to understand the morphology
of each cell population.
Through our research, we aim to uncover novel physical insights into the endosomal escape of LNPs and
determine the optimal membrane properties of LNPs to enhance fusion in living systems. This will lead to the
development of RNA delivery vehicles that are significantly more effective in delivering their cargo.

## Key facts

- **NIH application ID:** 10798629
- **Project number:** 3R01GM143723-02S1
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
- **Principal Investigator:** Cecilia Maria Leal
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $167,855
- **Award type:** 3
- **Project period:** 2022-06-10 → 2026-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10798629

## Citation

> US National Institutes of Health, RePORTER application 10798629, Endosomal escape of lipid-based nanoparticles comprising Gaussian curvature lipids (3R01GM143723-02S1). Retrieved via AI Analytics 2026-05-30 from https://api.ai-analytics.org/grant/nih/10798629. Licensed CC0.

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