{
  "abstract": "Background: Macrophages must coordinate immune activation with maintenance of cellular cholesterol homeostasis, yet the transcriptional mechanisms integrating these processes remain incompletely understood. Activating transcription factor 3 (ATF3) suppresses inflammatory and interferon responses and has been implicated in macrophage lipid metabolism, suggesting a role in coordinating immune and sterol-responsive transcriptional programs. Methods: Wild-type and ATF3-deficient bone marrow-derived macrophages were analyzed by bulk RNA sequencing following Toll-like receptor stimulation and under conditions of sterol loading or depletion. Cholesterol content, cytokine production, and sterol-responsive gene expression were quantified. Human metabolic dysfunction-associated steatotic liver disease (MASLD) datasets and Kupffer cell-specific ATF3 knockout mice were used to assess disease relevance in vivo. Results: ATF3 deficiency enhanced inflammatory and interferon-responsive transcriptional programs while suppressing a sterol-responsive gene module enriched for SREBP2 target genes. This phenotype was conserved across diverse Toll-like receptor stimuli and accompanied by persistent Ch25h upregulation. Despite exhibiting elevated cellular cholesterol, ATF3-deficient macrophages retained transcriptional responsiveness to sterol loading and depletion while demonstrating reduced induction of SREBP2-dependent genes across conditions. IFNAR blockade reduced interferon-responsive gene expression but did not normalize Ch25h or sterol-responsive gene expression. In contrast, CRISPR-mediated reduction of Ch25h partially restored sterol-responsive gene expression in inflammatory ATF3-deficient macrophages. In human MASLD, hepatic ATF3 expression positively correlated with disease severity. In mice, ATF3 was preferentially induced in Kupffer cells during metabolic stress, and Kupffer cell-specific deletion of ATF3 increased liver injury and Kupffer cell lipid accumulation. Conclusions: ATF3 supports sterol-responsive transcriptional programs while restraining inflammatory activation in macrophages. ATF3 loss results in elevated cellular cholesterol, impaired SREBP2-dependent gene expression, and persistent derepression of Ch25h, which contributes to but does not fully explain the sterol phenotype. These findings identify ATF3 as a coordinator of macrophage adaptation to inflammatory and metabolic stress and support a protective role for Kupffer cell ATF3 during metabolic liver disease.",
  "abstract_shown": true,
  "affiliations": [
    "Cincinnati Children's Hospital Medical Center"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 9,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "T. D. Troutman"
    },
    {
      "name": "A. M. Pearson"
    },
    {
      "name": "Y. Barudi"
    },
    {
      "name": "J. Kang"
    },
    {
      "name": "L. Williams"
    },
    {
      "name": "C. N. Vallez"
    },
    {
      "name": "C. J. Ulanowicz"
    },
    {
      "name": "N. S. Gokhale"
    },
    {
      "name": "S. Divanovic"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/e5ghxaehzp87s.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.11.03.764124",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.11.03.764124v1.full",
  "id": "e5ghxaehzp87s",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.11.03.764124v1",
  "language": null,
  "licence": "CC-BY-NC-ND",
  "licence_label": "CC BY-NC-ND",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "immunology"
    ]
  ],
  "openalex": null,
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.11.03.764124v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-11-03",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/e5ghxaehzp87s.ris",
  "sources": [
    {
      "first_seen": "2026-11-09T12:21:46Z",
      "native_id": "10.64898/2026.11.03.764124",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.11.03.764124v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:immunology",
      "subfield": "immunology"
    }
  ],
  "title": "ATF3 Preserves Macrophage Cholesterol Homeostasis during Metabolic Stress",
  "title_text": "ATF3 Preserves Macrophage Cholesterol Homeostasis during Metabolic Stress",
  "type": "preprint",
  "updated_at": "2026-11-09T12:21:46Z",
  "url": "https://research.draconitic.com/paper/e5ghxaehzp87s",
  "venue": "bioRxiv",
  "versions": []
}