Guselkumab binding to CD64 + IL-23-producing myeloid cells enhances potency for neutralizing IL-23 signaling

Kacey L. Sachen ^1* Deepa Hammaker ¹ Indra Sarabia ¹ Brian Stoveken ² John Hartman ² Kristin L Leppard ² Nicholas A Manieri ² Phuc Bao ¹ Carrie Greving ¹ Eilyn R Lacy ² Matthew DuPrie ² Joshua Wertheimer ² Janise D Deming ¹ Joseph Brown ¹ Amy Hart ² He (Hurley) Li ² Tom C. Freeman ² Brice Keyes ¹ Kristen Kohler ¹ Ian White ² Nathan Karpowich ² Ruth Steele ² M Merle Elloso ³ Steven Fakharzadeh ³ Kavitha Goyal ³ Frédéric Lavie ⁴ Maria T Abreu ⁵ Matthieu Allez ⁶ Raja Atreya ⁷ Robert Bissonnette ⁸ Kilian Eyerich ^10,9 James G Krueger ¹¹ Dennis McGonagle ¹² Iain McInnes ¹³ Christopher Ritchlin ¹⁴ Anne M Fourie ¹

• ¹ Janssen Research & Development, LLC, San Diego, California, United States
• ² Janssen Research & Development, LLC, Spring House, PA, United States
• ³ Janssen Scientific Affairs, LLC, Immunology, Horsham, PA, United States
• ⁴ The Janssen Pharmaceutical Companies of Johnson & Johnson, Paris, France
• ⁵ University of Miami, Leonard M. Miller School of Medicine, Miami, FL, United States
• ⁶ Hôpital Saint-Louis, Université Paris Cité, Paris, France
• ⁷ Department of Medicine 1, Erlangen University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
• ⁸ Innovaderm Research Inc, Montréal, Québec, Canada
• ⁹ Medical Center, University of Freiburg, Freiburg, Germany
• ¹⁰ Karolinska Institute, Department of Medicine – Division of Dermatology and Venereology, Stockholm, Sweden
• ¹¹ The Rockefeller University, Laboratory for Investigative Dermatology, New York, NY, United States
• ¹² University of Leeds, Leeds Biomedical Research Centre, Leeds, United Kingdom
• ¹³ University of Glasgow, College of Medical, Veterinary, and Life Sciences, Glasgow, United Kingdom
• ¹⁴ University of Rochester, Center for Musculoskeletal Research, Allergy, Immunology, and Rheumatology Division, Rochester, NY, United States

IL-23 is implicated in the pathogenesis of immune-mediated inflammatory diseases, and myeloid cells that express Fc gamma receptor 1 (FcγRI or CD64) on their surface have been recently identified as a primary source of IL-23 in inflamed tissue. Our complementary analyses of transcriptomic datasets from psoriasis and IBD showed increased expression of CD64 and IL-23 transcripts in inflamed tissue, and greater abundance of cell types with co-expression of CD64 and IL-23. These findings led us to explore potential implications of CD64 binding on the function of IL-23-targeting monoclonal antibodies (mAbs). Guselkumab and risankizumab are mAbs that target the IL-23p19 subunit. Guselkumab has a native Fc domain while risankizumab contains mutations that diminish binding to FcRs. In flow cytometry assays, guselkumab, but not risankizumab, showed Fcmediated binding to CD64 on IFNγ-primed monocytes. Guselkumab bound CD64 on IL-23producing inflammatory monocytes and simultaneously captured IL-23 secreted from these cells. Guselkumab binding to CD64 did not induce cytokine production. In live-cell confocal imaging of CD64 + macrophages, guselkumab, but not risankizumab, mediated IL-23 internalization to low-pH intracellular compartments. Guselkumab and risankizumab demonstrated similar potency for inhibition of IL-23 signaling in cellular assays with exogenous addition of IL-23. However, in a coculture of IL-23-producing CD64 + THP-1 cells with an IL-23-responsive reporter cell line, guselkumab demonstrated Fc-dependent enhanced potency compared to risankizumab for inhibiting IL-23 signaling. These in vitro data highlight the potential for guselkumab binding to CD64 in inflamed tissue to contribute to the potent neutralization of IL-23 at its cellular source.