# Identification and characterization of small open reading frames translated during inflammation

> **NIH NIH F31** · UNIVERSITY OF CALIFORNIA SANTA CRUZ · 2024 · $44,836

## Abstract

PROJECT SUMMARY
Next generation sequencing technologies have greatly expanded the size of the known transcriptome. Many
newly discovered transcripts are classified as long noncoding RNAs (lncRNAs) which are assumed to influence
phenotype through sequence and structure and not via translated protein products, despite the vast majority of
them harboring short open reading frames (sORFs). Recent advances have demonstrated that the noncoding
designation is incorrect in many cases and that sORF-encoded peptides (SEPs also called micropeptides)
translated from these transcripts are important contributors to diverse biological processes including
inflammation and cell viability. An appropriate inflammatory response is critical for host defense against
pathogens, but chronic inflammation is associated with many diseases. Macrophages play a significant role in
both initiating and resolving inflammation and understanding their part in this process is scientifically and
practically important. One long studied - yet not fully understood - model of macrophage proinflammatory
polarization involves lipopolysaccharide (LPS) activation of toll-like receptor 4 (TLR4). Following detection of
LPS, a signaling cascade initiates leading to the translocation of transcription factor NFkB to the nucleus. This
is followed by increased expression of established inflammatory cytokine and interferon genes. However, this
also results in changes in expression of many unstudied lncRNAs. In addition to changes in transcription,
changes in translation also follow inflammatory stimulation and these alterations have been observed to
increase translation of “noncoding” regions in some cases. Indeed our lab and others have observed dramatic
changes in associations between lncRNAs and polysomes following LPS stimulation in mouse macrophages.
Therefore, the central hypothesis of this proposal is that translation of lncRNAs produce SEPs that
play important roles in the TLR4-NFkB inflammatory response and in macrophage viability. To test this
hypothesis, I present a strategy for screening lncRNA sORFs with evidence of coding potential in mouse
macrophages. The screen will make use of macrophage cell lines with a NFkB-GFP reporter and CRISPR-
Cas9 casssette. Secondly, I propose a biomolecular pipeline for mechanistically characterizing a selection of
novel SEPs. This work has the potential to identify many novel SEPs that are important for regulating the
inflammatory response. This would further our understanding of a model inflammatory pathway and could help
identify novel peptides with therapeutic potential or as therapeutic targets.

## Key facts

- **NIH application ID:** 10915440
- **Project number:** 5F31AI179201-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA SANTA CRUZ
- **Principal Investigator:** Eric Malekos
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $44,836
- **Award type:** 5
- **Project period:** 2023-08-01 → 2026-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10915440

## Citation

> US National Institutes of Health, RePORTER application 10915440, Identification and characterization of small open reading frames translated during inflammation (5F31AI179201-02). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10915440. Licensed CC0.

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