# Project 2: Cholesterol Regulation of Inflammatory Macrophages in Atherosclerosis

> **NIH NIH P01** · LA JOLLA INSTITUTE FOR IMMUNOLOGY · 2022 · $42,993

## Abstract

Project 2 Summary
Atherosclerosis is the primary cause of cardiovascular disease (CVD), which manifests in myocardial infarction
and stroke, a major source of mortality and morbidity in the US. Hypercholesterolemia and chronic
inflammation are leading causative factors in the development of atherosclerosis. Many inflammatory receptors
and other proteins involved in the inflammatory response localize and become activated in cholesterol-rich
membrane microdomains, often designated as lipid rafts, which provide a solid platform for protein assembly
within a liquid membrane. Here, we propose the concept of inflammarafts, enlarged lipid rafts hosting
assembled inflammatory signaling complexes, to help formalize the lipid raft-centric view of inflammation and
its control by cholesterol metabolism. Cholesterol depletion from inflammarafts in activated cells disrupts
inflammatory signaling, and the approaches that allow for targeting cholesterol removal selectively to
inflammatory cells may be used for atheroprotection. In the context of atherosclerosis, monocytes and
macrophages develop long-term adaptation, or trained immunity, which results in sustained inflammatory
phenotypes and includes epigenetic memory, as well as transcriptional and metabolic alterations leading to a
chronic inflammatory state. In this application, we will test the hypotheses that inflammarafts serve as a
signaling platform that mediates cell reprogramming and that trained immune cells sustain lipid rafts to
maintain the inflammatory response. We propose that this positive feedback mechanism between
inflammarafts and metabolic, gene expression and epigenetic alterations plays a major role in the development
of atherosclerosis. Specifically, to test the hypothesis that inflammarafts serve as gatekeepers and effectors of
trained immunity in atherosclerosis, we will use inflammaraft assays to image and quantify lipid rafts,
accessible cholesterol, membrane order, TLR4 dimerization and the assembly of other receptor complexes.
RNA-seq, ATAC-seq, ChIPseq and metabolomics will be used to characterize trained immunity. To modulate
ligand-dependent inflammaraft assembly, we will neutralize oxidized lipid DAMPs, abundant in plaques, in
transgenic mice that overexpress oxidation-specific antibodies. APOA1, AAV-AIBP (apoA-I binding protein)
and cyclodextrin interventions will be used for systemic and targeted depletion of cholesterol. Conversely,
cholesterol and inflammarafts will be increased in inducible, macrophage-specific ABCA1/ABCG1 knockout
mice. Statins/mevalonate and desmosterol mimetics will be used to modulate macrophage reprogramming.
Importantly, we will evaluate the diagnostic and prognostic power of inflammaraft phenotyping of blood
leukocytes in CVD. By using the resources of PPG’s clinical and single cell protein and RNA sequencing cores,
we will test the hypotheses that the abundance of inflammarafts and raft-dependent receptor assemblies in
peripheral blood monocytes and...

## Key facts

- **NIH application ID:** 10334095
- **Project number:** 2P01HL136275-06
- **Recipient organization:** LA JOLLA INSTITUTE FOR IMMUNOLOGY
- **Principal Investigator:** Yury Miller
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $42,993
- **Award type:** 2
- **Project period:** 2017-08-01 → 2022-06-02

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10334095

## Citation

> US National Institutes of Health, RePORTER application 10334095, Project 2: Cholesterol Regulation of Inflammatory Macrophages in Atherosclerosis (2P01HL136275-06). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10334095. Licensed CC0.

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