Targeting RIPK3 oligomerization blocks necroptosis without inducing apoptosis

Receptor-interacting serine/threonine-protein kinase 3 (RIPK3) is a central protein in Necroptosis with great potential as a target for treating necroptosis-associated diseases, such as Crohn's disease. However, blockade of RIPK3 kinase activity leads to unexpected RIPK3-initiated Apoptosis. Herein, we found that PP2, a known Src Inhibitor, inhibits TNF-α-induced Necroptosis without initiating Apoptosis. Further investigation showed that PP2 acts as an inhibitor of not only Src but also RIPK3. PP2 does not disturb the integrity of the RIPK1-RIPK3-mixed lineage kinase domain-like pseudokinase (MLKL) necroptosome or the autophosphorylation of RIPK3 at T231/S232 but disrupts RIPK3 oligomerization, thereby impairing the phosphorylation and oligomerization of MLKL. These results demonstrate the essential role of RIPK3 oligomerization in Necroptosis and suggest a potential RIPK3 oligomerization-targeting strategy for therapeutic development.