# Innate Immunity, Cholesterol, and NASH Pathogenesis

> **NIH NIH R01** · UNIVERSITY OF SOUTHERN CALIFORNIA · 2023 · $371,250

## Abstract

PROJECT SUMMARY /ABSTRACT
Non-alcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease, affecting at least
a quarter of the world’s population, and therefore highly relevant to the NIH mission. From steatosis to fibrosis,
hepatic inflammation drives disease progression, and this study will elucidate the central roles of cholesterol.
The lack of evidence-based, FDA-approved treatment options for NASH underscores the need for further
research into understanding disease pathogenesis and identifying potential targets. Our compelling preliminary
and recently-published findings demonstrate innate immune cells (neutrophils and macrophages) are central to
the pathogenesis and progression of NAFLD. The overarching hypothesis of this proposal is that dysregulation
of innate immunity and inflammation is a dominant, indispensable feature of NAFLD, largely mediated by
cholesterol. We will test this hypothesis through completion of three Specific Aims performed in parallel using
murine NAFLD models and samples from patients with varying severity of NASH. In SA1, the PI will
comprehensively characterize fresh peripheral neutrophils from patients with different stages of NASH. This will
include NETosis, how cholesterol-induced NETs license responses from macrophages and differential effect of
neutrophils on liver endothelial cells. Murine models will focus on targeting of NETs to attenuate hepatic injury
in NASH. We will use the FPC diet, rich in fructose, palmitate, trans-fat, and cholesterol (1.2% by wt), which
has been shown to induce hepatic fibrosis-associated parameters within 16 weeks. SA2 will use live mice fed
different FPC-based diets with isolation of hepatic tissue by ultrasound-guided fine-needle aspiration, followed
by single-cell RNA sequencing (scRNA-seq) to measure genome-wide expression of macrophages from
progressing or regressing diet-induced NASH. Our data demonstrate that withdrawal of high cholesterol in the
diet downregulates macrophage-specific transcription as early as 4 weeks. The proposed experimental plan will
provide unprecedented insights into how dietary cholesterol affects different hepatic macrophage populations.
We will also characterize the functional consequences of different hepatic macrophages on co-cultured hepatic
stellate cells (HSCs). Moreover, we will use targeted sequencing of promoter regions in macrophage genes in
order to define key adaptive changes in epigenetic regulation. Finally, in SA3, the PI will focus on integrating
these robust analyses and utilizing Cre-Lox system to generate mice with myeloid-specific targeted mutants of
genes. As we have found that osteopontin (OPN)-encoding SPP1 expression was strongly induced in
macrophages in NASH, we will test an OPN nanoparticle in prevention and reversal studies of diet-induced
NASH. Mice carrying the Col1a1-GFP transgene will be used to directly assess the effect of different therapeutic
strategies on activation, differentiation, an...

## Key facts

- **NIH application ID:** 10595671
- **Project number:** 5R01DK117004-04
- **Recipient organization:** UNIVERSITY OF SOUTHERN CALIFORNIA
- **Principal Investigator:** Lucy Mary Golden
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $371,250
- **Award type:** 5
- **Project period:** 2020-01-20 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10595671

## Citation

> US National Institutes of Health, RePORTER application 10595671, Innate Immunity, Cholesterol, and NASH Pathogenesis (5R01DK117004-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10595671. Licensed CC0.

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