Study On Chronic Diseases Caused By Nanoparticles Based On NLRP3 Inflammasome Activation And Its Application In Tumor Therapy

NLRP3 inflammasome is an intracellular multiprotein complex assembled in response to various stimuli,which controls caspase-1 activation and regulates the secretion of IL-1β and IL-18.Activation of NLRP3 inflammasome has been implicated in many chronic diseases,including pulmonary fibrosis,Alzheimer’s disease,atherosclerosis,obesity,cancer and so on.At present,a variety of nanomaterials have been found to activate NLRP3 inflammasome through lysosome rupture,reactive oxygen species production,potassium ion efflux and other pathways,which are closely related to its unique physical and chemical properties.However,there are a large number of nano-scale particles with complex physical and chemical properties in the environment and food,which may enter deeply lung or brain tissues of human body through breathing or swallowing,and cause a variety of chronic respiratory system diseases or brain diseases by activating NLRP3 inflammasome.At the same time,activation of the high-potency and potent NLRP3 inflammasome also acts as an immune adjuvant,binding to antigens and recruiting more CD8+and CD4+T cells to attack tumor cells.Therefore,the design of functional nanomaterials that can efficiently activate NLRP3 inflammasome may form a promising cancer vaccine for the treatment of cancer.In this paper,the chronic fibrosis effect of atmospheric fine particles(PM2.5)and rare earth biotransformed particles was studied by focusing on the biological effect of NLRP3 inflammasome activation,and functional nanomaterials were designed and constructed to activate NLRP3 inflammasome to play the role of cancer vaccine and treat tumors.The main contents are as follows:(1)To clarify the composition of PM2.5 and evaluate its ability to activate NLRP3 inflammasome and induce pulmonary fibrosis at the cellular and animal levels.The results show that the main chemical components of PM2.5 are water-soluble ions,carbon species,metal elements and so on.PM2.5 can activate NLRP3 inflammasome through lysosome rupture,K+efflux and ROS production.Oral inhalation of PM2.5 in the lungs can trigger the activation of NLRP3 inflammasome,leading to pulmonary fibrosis.In order to further explore the relationship between its properties and activity,the physical and chemical properties of PM2.5 are changed by metal chelation(MetalsPM2.5),organic extraction(PAHsPM2.5),ROS consumption(ROSPM2.5),charge neutralization(ChargePM2.5)and particles dispersion(SizePM2.5).MetalsPM2.5,PAHsPM2.5 and ROSPM2.5 effectively inhibit IL-1β release by reducing mitochondrial ROS production,while ChargePM2.5 further enhances lysosomal rupture and cathepsin B release to increase IL-1β release.Additionally,PAHsPM2.5 also reduced K+efflux to decrease IL-1β release.(2)It has been established that PM2.5 can promote the initiation of Alzheimer’s disease(AD)by activating NLRP3 inflammasome and releasing ASC specks.It was found that PM2.5 can activate NLRP3 inflammasome of neuromicroglia(BV2)through a variety of ways,and release ASC specks,which can interact with Aβ,accelerate Aβ aggregation,and promote neuronal apoptosis.After APP/PS1 mice were intranasally injected with PM2.5 for 13 weeks,ASC specks release and Aβ aggregation of BV2 were observed in the hippocampus of mice.In addition,high concentrations of PM2.5 led to a significant reduction in the number of neurons and spatial memory deficits in APP/PS1 mice.These results indicate that PM2.5 has a strong correlation with AD.(3)Soluble La(Ⅲ)species in rare earth microfertilizer can be transformed into insoluble La(Ⅲ)particles under physiological conditions,which can trigger chronic fibrosis of liver and kidney by activating NLRP3 inflammasome.It was found that the insoluble La(Ⅲ)particles formed under physiological conditions were in nanometer or micron size,and the main components were La2(CO3)3 and LaPO4.The main components of these particles undergo biotransformation after contact with cells.Insoluble La(Ⅲ)particles can adhere to the extracellular membrane or be endocytosed into cells and activate NLRP3 inflammasome through K+efflux and lysosomal rupture mechanisms.This is mainly because these insoluble La(Ⅲ)particles located outside the cell membrane can reduce the unsaturated fatty acids on the cell membrane,destroy the stability of the membrane,and lead to K+efflux.And those particles that internalize into the cell deplete the phospholipids in the lysosome membrane,causing the lysosome to break down.In mice drinking soluble La(Ⅲ)for 90 days to simulate tea drinking in healthy adults,collagen deposition in liver and kidney was found,leading to chronic fibrosis,which may be closely related to the formation of insoluble La(Ⅲ)particles.(4)Acidic and GSH-responsive,variably sized nanoparticles(CSMP NPs)were designed and prepared to provide DCs cells with nanovaccines by activating NLRP3 inflammasome and killing tumor cells as antigens for tumor immunotherapy.CSMP NPs consists of copper-manganese nanoclusters loaded with POM.In acidic DCs lysosomes,CSMP NPs can aggregate into micron structures that disrupt lysosome integrity,leading to the release of cathepsin B,which activates the NLRP3 inflammasome and releases large amounts of IL-1β,thus acting as an excellent adjuvant.At the same time,CSMP NPs is degraded gradually under the condition of high GSH in tumor cells,so as to penetrate deeply into tumor tissues,exert the effect of CDT and PTT,kill tumor cells,provide antigens for DCs,and thus form cancer vaccines in situ.Cancer vaccine formation encourages DCs to mature and migrate to lymph nodes for antigen presentation,recruiting more CD8+and CD4+T cells for robust immunotherapy.(5)A novel CCMZ nanosystem with enhanced chemodynamic therapy(CDT)and immunotherapy stimulation was designed and prepared for tumor therapy.PEGylated CuMoOx loaded with zinc porphyrins(ZP)produced CCMZ NPs,which enhanced CDT therapeutic effect by inhibiting HO-1 activity and depleting GSH.More importantly,the GSH responsiveness of CCMZ NPs makes it easy to penetrate deeply into tumor tissues,addressing the problem of poor drug penetration.At high levels of GSH,CCMZ degradation can achieve deep tumor penetration,resulting in CuMoOx shell degradation.At the same time,GSH itself is depleted,transforming the reducing environment into a continuous oxidation environment,resulting in the degradation of internal Cu NPs.The ZP released by CCMZ NPs can inhibit HO-1 activity and provide a good microenvironment for CDT.The release of Cu and Mo ions can convert H2O2 into ·OH and kill tumor cells more effectively.In addition,CCMZ NPs also plays an immune-vaccine-like role,which can recruit different immune cells for anti-tumor immunotherapy.In vitro and in vivo studies have confirmed the CDT enhancement properties of CCMZ NPs,providing a new strategy for improving the efficacy of CDT.