Human genetics guides the discovery of CARD9 inhibitors with anti-inflammatory activity

Jason S Rush¹, Joshua D Wertheimer², Steven D Goldberg³, Donald Raymond¹, Mateusz Szuchnicki¹, Andrew J Baltus², Jeff Branson², Christopher F Stratton², Aaron N Patrick², Ruth Steele², Suraj Adhikary², Amanda Del Rosario², Annie Liu², Noah J Gomersall², Michael Chung³, Matthew J Ranaghan¹, Xiebin Gu⁴, Marta Brandt⁵, Zhifang Cao⁵, Adrian Bebenek⁶, Blayne A Oliver⁴, Kasper Hoebe², Lawrence M Szewczuk², Jennifer D Venable³, Daniel B Graham⁷, Jennifer Towne³, Ramnik J Xavier⁸

Affiliations

¹ Center for the Development of Therapeutics, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
² Johnson & Johnson Innovative Medicine, Spring House, PA 19477, USA.
³ Johnson & Johnson Innovative Medicine, San Diego, CA 92121, USA.
⁴ Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Molecular Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
⁵ Center for the Development of Therapeutics, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Molecular Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
⁶ Center for the Development of Therapeutics, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA; Chemical Biology PhD Program, Harvard Medical School, Boston, MA 02115, USA.
⁷ Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Molecular Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
⁸ Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Molecular Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: rxavier@broadinstitute.org.

PMID: 41547356
PMCID: PMC12817910 (available on 2027-01-16)
DOI: 10.1016/j.cell.2025.12.013

Human genetics guides the discovery of CARD9 inhibitors with anti-inflammatory activity

Jason S Rush et al. Cell. 2026.

. 2026 Jan 16:S0092-8674(25)01426-6.

doi: 10.1016/j.cell.2025.12.013. Online ahead of print.

Authors

Affiliations

¹ Center for the Development of Therapeutics, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
² Johnson & Johnson Innovative Medicine, Spring House, PA 19477, USA.
³ Johnson & Johnson Innovative Medicine, San Diego, CA 92121, USA.
⁴ Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Molecular Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
⁵ Center for the Development of Therapeutics, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Molecular Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
⁶ Center for the Development of Therapeutics, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA; Chemical Biology PhD Program, Harvard Medical School, Boston, MA 02115, USA.
⁷ Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Molecular Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
⁸ Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Molecular Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: rxavier@broadinstitute.org.

PMID: 41547356
PMCID: PMC12817910 (available on 2027-01-16)
DOI: 10.1016/j.cell.2025.12.013

Abstract

Human genetic association studies highlight key genes involved in disease pathology, yet targets identified by these analyses often fall outside the traditional definitions of druggability. A rare truncated variant of the scaffold protein CARD9 is linked with protection from Crohn's disease, prompting us to pursue the development of inhibitors that might similarly modulate innate inflammatory responses. Using a phased approach, we first identified a ligandable site on CARD9 using a structurally diverse DNA-encoded library and defined this site in detail through X-ray crystallography. Building upon this, a subsequent ligand displacement screen identified additional molecules that uniquely engage CARD9 and prevent its assembly into scaffolds needed to nucleate a signalosome for downstream nuclear factor κB (NF-κB) induction. These inhibitors suppressed inflammatory cytokine production in dendritic cells and a humanized CARD9 mouse model. Collectively, this study illustrates a strategy for leveraging protective human genetic variants and chemical biology to tackle challenging targets for dampening inflammation.

Keywords: Crohn's disease; binder first; coiled coil; drug discovery; fungal immunity; immune-mediated disease; innate immunity; signalosome; small molecule inhibitor.

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Conflict of interest statement

Declaration of interests R.J.X. is board director at MoonLake Immunotherapeutics, co-founder of Convergence Bio, consultant to Nestlé, and a member of the scientific advisory board at Magnet BioMedicine; these organizations had no roles in this study.

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Human genetics guides the discovery of CARD9 inhibitors with anti-inflammatory activity

Affiliations

Human genetics guides the discovery of CARD9 inhibitors with anti-inflammatory activity

Authors

Affiliations

Abstract

Conflict of interest statement

References

Grants and funding

LinkOut - more resources

Full Text Sources