Directly targeting ASC by lonidamine alleviates inflammasome-driven diseases

by Selleckchem | Mar 26 2023

Background: Dysregulated activation of the inflammasome is involved in various human diseases including acute cerebral ischemia, multiple sclerosis and sepsis. Though many inflammasome inhibitors targeting NOD-like receptor protein 3 (NLRP3) have been designed and developed, none of the inhibitors are clinically available. Growing evidence suggests that targeting apoptosis-associated speck-like protein containing a CARD (ASC), the oligomerization of which is the key event for the assembly of inflammasome, may be another promising therapeutic strategy. Lonidamine (LND), a small-molecule inhibitor of glycolysis used as an antineoplastic drug, has been evidenced to have anti-inflammation effects. However, its anti-inflammatory mechanism is still largely unknown.

Methods: Middle cerebral artery occlusion (MCAO), experimental autoimmune encephalomyelitis (EAE) and LPS-induced sepsis mice models were constructed to investigate the therapeutic and anti-inflammasome effects of LND. The inhibition of inflammasome activation and ASC oligomerization by LND was evaluated using western blot (WB), immunofluorescence (IF), quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA) in murine bone marrow-derived macrophages (BMDMs). Direct binding of LND with ASC was assessed using molecular mock docking, surface plasmon resonance (SPR), and drug affinity responsive target stability (DARTS).

Results: Here, we find that LND strongly attenuates the inflammatory injury in experimental models of inflammasome-associated diseases including autoimmune disease-multiple sclerosis (MS), ischemic stroke and sepsis. Moreover, LND blocks diverse types of inflammasome activation independent of its known targets including hexokinase 2 (HK2). We further reveal that LND directly binds to the inflammasome ligand ASC and inhibits its oligomerization.

Conclusions: Taken together, our results identify LND as a broad-spectrum inflammasome inhibitor by directly targeting ASC, providing a novel candidate drug for the treatment of inflammasome-driven diseases in clinic.

Comments：

In this study, the authors investigated the potential of lonidamine (LND), a small-molecule inhibitor of glycolysis used as an antineoplastic drug, as a therapeutic agent for inflammasome-associated diseases. They found that LND strongly attenuates the inflammatory injury in experimental models of inflammasome-associated diseases including autoimmune disease-multiple sclerosis (MS), ischemic stroke and sepsis. Furthermore, LND blocks diverse types of inflammasome activation independent of its known targets including hexokinase 2 (HK2).

The authors further revealed that LND directly binds to the inflammasome ligand apoptosis-associated speck-like protein containing a CARD (ASC) and inhibits its oligomerization, which is the key event for the assembly of inflammasome. These findings suggest that LND is a broad-spectrum inflammasome inhibitor by directly targeting ASC, providing a novel candidate drug for the treatment of inflammasome-driven diseases in clinic.

The authors used several experimental approaches including middle cerebral artery occlusion (MCAO), experimental autoimmune encephalomyelitis (EAE), and LPS-induced sepsis mice models to investigate the therapeutic and anti-inflammasome effects of LND. They also evaluated the inhibition of inflammasome activation and ASC oligomerization by LND using western blot (WB), immunofluorescence (IF), quantitative polymerase chain reaction (qPCR), and enzyme-linked immunosorbent assay (ELISA) in murine bone marrow-derived macrophages (BMDMs). Finally, they assessed the direct binding of LND with ASC using molecular mock docking, surface plasmon resonance (SPR), and drug affinity responsive target stability (DARTS).

Overall, this study provides important insights into the mechanism of action of LND as an inflammasome inhibitor and suggests that it may have potential as a therapeutic agent for inflammasome-associated diseases. However, further studies are needed to evaluate the safety and efficacy of LND in clinical settings.