Bruton's Tyrosine Kinase Inhibition Attenuates the Cardiac Dysfunction Caused by Cecal Ligation and Puncture in Mice

Abstract

Sepsis is one of the most prevalent diseases in the world. The development of cardiac dysfunction in sepsis results in an increase of mortality. It is known that Bruton's tyrosine kinase (BTK) plays a role in toll-like receptor signaling and NLRP3 inflammasome activation, two key components in the pathophysiology of sepsis and sepsis-associated cardiac dysfunction. In this study we investigated whether pharmacological inhibition of BTK (ibrutinib 30 mg/kg and acalabrutinib 3 mg/kg) attenuates sepsis associated cardiac dysfunction in mice. 10-week old male C57BL/6 mice underwent CLP or sham surgery. One hour after surgery mice received either vehicle (5% DMSO + 30% cyclodextrin i.v.), ibrutinib (30 mg/kg i.v.), or acalabrutinib (3 mg/kg i.v.). Mice also received antibiotics and an analgesic at 6 and 18 h. After 24 h, cardiac function was assessed by echocardiography in vivo. Cardiac tissue underwent western blot analysis to determine the activation of BTK, NLRP3 inflammasome and NF-κB pathway. Serum analysis of 33 cytokines was conducted by a multiplex assay. When compared to sham-operated animals, mice subjected to CLP demonstrated a significant reduction in ejection fraction (EF), fractional shortening (FS), and fractional area change (FAC). The cardiac tissue from CLP mice showed significant increases of BTK, NF-κB, and NLRP3 inflammasome activation. CLP animals resulted in a significant increase of serum cytokines and chemokines (TNF-α, IL-6, IFN-γ, KC, eotaxin-1, eotaxin-2, IL-10, IL-4, CXCL10, and CXCL11). Delayed administration of ibrutinib and acalabrutinib attenuated the decline of EF, FS, and FAC caused by CLP and also reduced the activation of BTK, NF-κB, and NLRP3 inflammasome. Both ibrutinib and acalabrutinib significantly suppressed the release of cytokines and chemokines. Our study revealed that delayed intravenous administration of ibrutinib or acalabrutinib attenuated the cardiac dysfunction associated with sepsis by inhibiting BTK, reducing NF-κB activation and the activation of the inflammasome. Cytokines associated with sepsis were significantly reduced by both BTK inhibitors. Acalabrutinib is found to be more potent than ibrutinib and could potentially prove to be a novel therapeutic in sepsis. Thus, the FDA-approved BTK inhibitors ibrutinib and acalabrutinib may be repurposed for the use in sepsis.

X-Linked Immunodeficient Mice With No Functional Bruton's Tyrosine Kinase Are Protected From Sepsis-Induced Multiple Organ Failure

“…They were subjected to a 12 h light and dark cycle with a temperature maintained at 19–23°C. Group sizes for each experiment were calculated following power calculations based on previous studies ( 14 ).…”

“…Cecal ligation and puncture (CLP) was performed in 10 week-old male CBA (wild type) or Xid mice as previously described ( 14 , 22 ). Mice were randomly assigned to undergo CLP or sham-operated surgery, the surgeon was blinded to the genotype of the mouse.…”

“…We have recently shown that the BTK inhibitors ibrutinib (first generation) and acalabrutinib (more selective, second generation) attenuate the systemic inflammation (“cytokine storm”) and the multiple organ failure caused by sepsis in mice ( 14 ). Ibrutinib is already approved for the use in chronic lymphatic leukemia, mantle cell lymphoma, Waldenstrom macroglobulinemia, and graft vs. host disease ( 15 ) and acalabrutinib in mantle cell lymphoma ( 16 ).…”

“…Although we have proposed that the inhibition of BTK is the key driver of the observed beneficial effects of BTK inhibitors in sepsis, it is possible that some of the well-known off-target effects of these compounds account for or, at least, contribute to the beneficial effects observed ( 14 ). For instance, we identified that both ibrutinib and acalabrutinib strongly inhibit five different kinases: BTK, Bmx, ErbB4, RIPK2, and TEC.…”

The hidden role of NLRP3 inflammasome in obesity‐related COVID‐19 exacerbations: Lessons for drug repurposing

“…NLRP3 is expressed in several cell types, as innate immunity, endothelial, haematopoietic, lung epithelial, kidney, and cardiac cells (Ratajczak & Kucia, 2020). We and others have previously demonstrated that NLRP3 inflammasome over-activation contributes to the pathogenesis of cardio-metabolic disorders (Mastrocola, Collino, et al, 2016;O'Riordan et al, 2019;Zuurbier et al, 2019), whereas NLRP3 deficiency results in reduced systemic inflammation, along with a decreased immune cell activation and improved insulin resistance (Grant & Dixit, 2013;Vandanmagsar et al, 2011). The NLRP3 inflammasome is also a crucial player in immune defence of the host against many pathogens, including viruses (Z.…”