Optimized protocols for studying the NLRP3 inflammasome and assessment of potential targets of CP-453,773 in undifferentiated THP1 cells

Guzova, Julia A.; Primiano, Michael J.; Jiao, Aiping; Stock, Jeffrey L.; Lee, Chiachin; Winkler, Aaron; Hall, J. Perry

doi:10.1016/j.jim.2019.02.002

Cited by 11 publications

(11 citation statements)

References 49 publications

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“…MCC950 dose-dependently downregulated IL-1b but did not block K + efflux, Ca 2+ flux, or ligand-independent direct NLRP3 and apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC) interactions. MCC950 also did not affect mitochondrial transporters ATP-binding cassette b7 and ATP-binding cassette b10 (Guzova et al, 2019). Further work by Pfizer (Primiano et al, 2016) showed efficacy in pulmonary models of inflammation, with only very weak off-target activity identified through commercially available screening panels.…”

Section: A Nod-like Receptor Family Pyrin Domain Containing 3 Inhibitorsmentioning

confidence: 94%

Pharmacological Inhibition of the Nod-Like Receptor Family Pyrin Domain Containing 3 Inflammasome with MCC950

2021

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Section: A Nod-like Receptor Family Pyrin Domain Containing 3 Inhibitorsmentioning

confidence: 94%

Pharmacological Inhibition of the Nod-Like Receptor Family Pyrin Domain Containing 3 Inflammasome with MCC950

2021

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“…MCC950, also known as CP-456773 or CRID3 ( Guzova et al., 2019 ; Kuwar et al., 2019 ), is widely known as the most potent and specific NLRP3 inhibitor ( Dai et al., 2021 ). It is often used as a positive control to compare the efficacy of novel NLRP3 inhibitors; for instance, the discovery of nitrostitu-quinazolin-4 (3H)-one 2 K to inhibit the activation of NLRP3 inflammatory body by occupying the ATP-binding site of NLRP3 protein ( Abdullaha et al., 2019 ).…”

Section: Specific Inhibitors Of Nlrp3 Inflammasomementioning

confidence: 99%

“…In recent years, it has been observed that MCC950 inhibits ASC oligomerization and inhibits the cleavage of caspase-1 (p20) (Chow et al, 2020;Dai et al, 2021). Guzova et al (2019) proposed that mitochondrial transporters ABCb7 and ABCb10 are the pharmacological targets of CP-453773; although this hypothesis has been proven incorrect, they found that undifferentiated THP1 cells can serve as a reliable and simplified model for investigating NLRP3 inflammasomes (Guzova et al, 2019). MCC950 was found to increase serum liver enzyme levels in a phase II clinical trial in rheumatoid arthritis; hence, it was not further developed .…”

Section: Inhibitors Of the Activation Stepmentioning

confidence: 99%

The Role of NLRP3 Inflammasome in Cerebrovascular Diseases Pathology and Possible Therapeutic Targets

et al. 2021

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Cerebrovascular diseases are pathological conditions involving impaired blood flow in the brain, primarily including ischaemic stroke, intracranial haemorrhage, and subarachnoid haemorrhage. The nucleotide-binding and oligomerisation (NOD) domain-like receptor (NLR) family pyrin domain (PYD)-containing 3 (NLRP3) inflammasome is a protein complex and a vital component of the immune system. Emerging evidence has indicated that the NLRP3 inflammasome plays an important role in cerebrovascular diseases. The function of the NLRP3 inflammasome in the pathogenesis of cerebrovascular diseases remains an interesting field of research. In this review, we first summarised the pathological mechanism of cerebrovascular diseases and the pathological mechanism of the NLRP3 inflammasome in aggravating atherosclerosis and cerebrovascular diseases. Second, we outlined signalling pathways through which the NLRP3 inflammasome participates in aggravating or mitigating cerebrovascular diseases. Reactive oxygen species (ROS)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), ROS/thioredoxin-interacting protein (TXNIP) and purinergic receptor-7 (P2X7R) signalling pathways can activate the NLRP3 inflammasome; activation of the NLRP3 inflammasome can aggravate cerebrovascular diseases by mediating apoptosis and pyroptosis. Autophagy/mitochondrial autophagy, nuclear factor E2-related factor-2 (Nrf2), interferon (IFN)-β, sirtuin (SIRT), and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) reportedly alleviate cerebrovascular diseases by inhibiting NLRP3 inflammasome activation. Finally, we explored specific inhibitors of the NLRP3 inflammasome based on the two-step activation of the NLRP3 inflammasome, which can be developed as new drugs to treat cerebrovascular diseases.

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“…In vitro assays using LPS, ATP and other activators are widely used to investigate stimulation of the NLRP3 inflammasome [6,[23][24][25][26]. These in vitro studies showed that NLRP3 inflammasome stimulation leads to the release of mature IL-1β [6,23,24].…”

Section: In Vitro Nlrp3 Inflammasome Stimulationmentioning

confidence: 99%

“…Since NLRP3 protein is an NLRP3 inflammasome component, this may indicate that EV NLRP3 protein levels reflect the status of the NLRP3 inflammasome in the parent cells. Stimulated NLRP3 inflammasome increases mature IL-1β levels and colchicine attenuates this release [26,27]. IL-1β is also released by the activation of other inflammasomes and cleavage of intracellular pro-IL-1β can be independent of inflammasomes [15].…”

Section: In Vitro Nlrp3 Inflammasome Stimulationmentioning

confidence: 99%

Colchicine reduces extracellular vesicle NLRP3 inflammasome protein levels in chronic coronary disease: A LoDoCo2 biomarker substudy

et al. 2021

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Background and aims: Colchicine reduces the risk of cardiovascular events in patients with coronary disease. Colchicine has broad anti-inflammatory effects and part of the atheroprotective effects have been suggested to be the result of NLRP3 inflammasome inhibition. We studied the effect of colchicine on extracellular vesicle (EV) NLRP3 protein levels and inflammatory markers, high sensitivity-CRP (hs-CRP) and interleukin (IL)-6, in patients with chronic coronary disease. Methods: In vitro, the NLRP3 inflammasome was stimulated in PMA-differentiated-and undifferentiated THP-1 cells. In vivo, measurements were performed in serum obtained from 278 participants of the LoDoCo2 trial, one year after randomization to colchicine 0.5 mg once daily or placebo. EVs were isolated using precipitation. NLRP3 protein presence in EVs was confirmed using iodixanol density gradient centrifugation. Levels of NLRP3 protein, hs-CRP and IL-6 were measured using ELISA. Results: In vitro, NLRP3 inflammasome stimulation showed an increase of EV NLRP3 protein levels. EV NLRP3 protein levels were lower in patients treated with colchicine (median 1.38 ng/mL), compared to placebo (median 1.58 ng/mL) (p = 0.025). No difference was observed in serum NLRP3 protein levels. Serum hs-CRP levels were lower in patients treated with colchicine (median 0.80 mg/L) compared to placebo (median 1.34 mg/L) (p < 0.005). IL-6 levels were lower in patients treated with colchicine (median 2.07 ng/L) compared to placebo (median 2.59 ng/L), although this was not statistically significant (p = 0.076). Conclusions: Colchicine leads to a reduction of EV NLRP3 protein levels. This indicates that inhibitory effects on the NLRP3 inflammasome might contribute to the atheroprotective effects of colchicine in coronary disease.

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Optimized protocols for studying the NLRP3 inflammasome and assessment of potential targets of CP-453,773 in undifferentiated THP1 cells

Cited by 11 publications

References 49 publications

Pharmacological Inhibition of the Nod-Like Receptor Family Pyrin Domain Containing 3 Inflammasome with MCC950

Pharmacological Inhibition of the Nod-Like Receptor Family Pyrin Domain Containing 3 Inflammasome with MCC950

The Role of NLRP3 Inflammasome in Cerebrovascular Diseases Pathology and Possible Therapeutic Targets

Colchicine reduces extracellular vesicle NLRP3 inflammasome protein levels in chronic coronary disease: A LoDoCo2 biomarker substudy

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