Alendronate Augments Lipid A–Induced IL-1α Release via Activation of ASC but Not Caspase-11

• Published in: Inflammation 2021-10, Vol.44 (5), p.2132-2141
• Main Authors: Tamai, Riyoko, Mashima, Izumi, Kiyoura, Yusuke
• Format: Article
• Language: English
• Subjects: Alendronate - administration & dosage; Alendronic acid; Animals; Apoptosis; Biomedical and Life Sciences; Biomedicine; Bisphosphonates; CARD Signaling Adaptor Proteins - metabolism; Caspase-11; Caspase-8; Caspases, Initiator; Cell culture; Cell death; Cell Line; Cell Survival - drug effects; Cell Survival - physiology; Cytosol; Dose-Response Relationship, Drug; Drug Synergism; Immunology; Inflammation; Interleukin-1alpha - metabolism; Internal Medicine; Kinases; Lipid A; Lipid A - administration & dosage; Lipids; Macrophages - drug effects; Macrophages - metabolism; MAP kinase; Necroptosis; Original Article; Pathology; Pharmacology/Toxicology; Protein kinase; Pyrin protein; Rheumatology
• ISSN: 0360-3997; 1573-2576
• DOI: 10.1007/s10753-021-01489-w

Nitrogen-containing bisphosphonates (NBPs), such as alendronate (ALN), are anti-bone-resorptive drugs that have inflammatory side effects. We previously reported that ALN augmented lipid A–induced interleukin (IL)-1β production and NOD-like receptor pyrin domain-containing-3 (NLRP3)/apoptosis-associated speck-like protein containing a CARD (ASC)-dependent cell death. The present study aimed to examine whether ALN augments lipid A–induced IL-1α release and necroptosis, which is induced by the activation of receptor-interacting protein kinase (RIPK) 3. Treatment of J774.1 cells with ALN augmented lipid A–induced IL-1α release, which was not inhibited by Ac-IETD-CHO, a caspase-8 inhibitor. ALN also activated mixed lineage kinase domain-like (MLKL), a key mediator of the necroptosis pathway, and upregulated the expression of caspase-11, a lipid A receptor. GSK′872, a RIPK3 inhibitor, suppressed the ALN-upregulated expression of caspase-11 and augmented lipid A–induced caspase-8 activation. Moreover, ALN induced the release of NLRP3 and ASC into culture supernatants. GSK′872, but not Ac-IETD-CHO, reduced the ALN-induced release of NLRP3, but not ASC, into culture supernatants, and reduced ALN-induced cell death, but not ALN-induced LDH release. Antibodies against NLRP3 and ASC upregulated caspase-11 expression in the cytosol by inhibiting ALN-induced cell death. However, pretreating cells with an antibody against ASC, but not NLRP3, before ALN addition also inhibited lipid A–induced IL-1α release. Pretreating cells with an antibody against caspase-11 before the addition of ALN or lipid A did not downregulate lipid A–induced production of IL-1α. Taken together, our findings suggest that ALN augments lipid A–induced IL-1α release via activation of ASC, but not caspase-11.