Category: Innate immunity
Cells of the immune system play a key role in the resolution of tissue injuries, inflammation and infections and in regulation of tissue homeostasis. Here we aimed to profile natural product-inspired drug discovery platform, developed by Janssen Incubator, using natural product scaffold-based chemical entities, to aid in identifying new chemical starting points with a disease-relevant mechanisms of action. We used 13 human primary cell-based assays, amongst which several assays target the immune system, to screen approximately 1,600 compounds.
Inflammatory assays utilized for screening include neutrophil migration, activation of B-cells, T-cells, macrophages, dendritic cells (DC) and synovial fibroblasts. Immune cells were isolated from healthy donor buffy coats and synovial fibroblasts were isolated from rheumatoid arthritis patients. Neutrophils were subjected to IL-8-mediated migration in a transwell system. Isolated B-cells were stimulated with CD40L and IL-4 to secrete IL-6. PBMCs were stimulated with anti-CD3/CD28 antibodies to induce IL-2 secretion by T-cells. For macrophage activation, PBMCs were stimulated with LPS to induce TNF-α secretion. Monocytes-derived DCs were stimulated with CD40L+enhancer to produce IL12p40. Synovial fibroblasts were stimulated with platelet-derived micro-particles to induce IL-6 secretion.
Assays were successfully validated using reference compounds and subsequently subjected to screening of the 1,600 compounds resulting in multiple new starting points for a hit-to-lead program. These screens also highlighted which assays could help identify unique, disease-relevant mechanisms of action.
Jamil Aarbiou– Group Leader, Charles River Laboratories
Krista Ouwehand– Group Leader, Charles River Laboratories
Julie Vann– Research Leader, Charles River Laboratories
David Fischer– Executive Director, Charles River Laboratories
Blandine Mille-Baker– Group Leader, Charles River Laboratories
Jeroen DeGroot– Senior Director, Charles River Laboratories