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Co-delivery of nigericin and decitabine using hexahistidine-metal nanocarriers for Pyroptosis-induced immunotherapeutics

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Acta Pharmaceutica Sinica B 2022年第12期12卷 4458-4471页

作者： Qiang Niu Yu Liu Yujing Zheng Ziwei Tang Yuna Qian Ruogu Qi Jianliang Shen Ping Zhao School of Chemistry and Chemical Engineering Guangdong Pharmaceutical UniversityGuangzhou 510006China State Key Laboratory of Ophthalmology Optometry and Vision ScienceSchool of Ophthalmology and OptometrySchool of Biomedical EngineeringWenzhou Medical UniversityWenzhou 325027China Wenzhou Institute University of Chinese Academy of SciencesWenzhou 325001China School of Medicine&Holistic Integrative Medicine Nanjing University of Chinese MedicineNanjing 210023China

Pyroptosis provides a new window for relieving the tumor immunosuppressive microenvironment(TIM)and promoting systemic immune responses for tumor treatments.However,gasdermin D(GSDMD),a key protein in the Pyroptosis process mediated by caspase-1,is low expressed in the majority of tumor cells and small-molecule inhibitors of DNA methylation suffer from nonspecific or single-function defects.To address these issues,hexahistidine(His6)-metal assembly(HmA)was employed as the drug delivery vector to load nigericin(Nig)and decitabine(DAC)affording a dual-drug delivery system(Nig^(+)DAC)@HmA.The(Nig^(+)DAC)@HmA nanoparticles are efficiently internalized by cells through endocytosis,easily escape from the lysosome,and are highly distributed in the tumor sites.DAC up-regulates the expression of GSDMD which is then cleaved by the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome and caspase-1 protein activated by Nig.Effective cancer cell Pyroptosis is thus achieved and induces a significant systemic antitumor immunity for impressive tumor suppression with negligible side effects in vivo.Our results suggest that such an easy-to-manipulate self-assembled nano-system(Nig^(+)DAC)@HmA provides a new anticancer path by enhancing Pyroptosis through reinforced inflammation.

关键词： Self-assembled nanoparticle Pyroptosis NLRP3 GSDMD Inflammation IL-8 Immunotherapy responses Solid tumor

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Mitochondrial transplantation confers protection against the effects of ischemic stroke by repressing microglial Pyroptosis and promoting neurogenesis

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Neural Regeneration Research 2024年第6期19卷 1325-1335页

作者： Li Sun Zhaoyan Zhao Jing Guo Yuan Qin Qian Yu Xiaolong Shi Fei Guo Haiqin Zhang Xude Sun Changjun Gao Qian Yang Department of Experimental Surgery The Second Affiliated Hospital of Air Force Medical University Department of Anesthesiology The Second Affiliated Hospital of Air Force Medical University

Transferring healthy and functional mitochondria to the lateral ventricles confers neuroprotection in a rat model of ischemia-reperfusion injury. Autologous mitochondrial transplantation is also beneficial in pediatric patients with cardiac ischemia-reperfusion injury. Thus, transplantation of functional exogenous mitochondria may be a promising therapeutic approach for ischemic disease. To explore the neuroprotective effect of mitochondria transplantation and determine the underlying mechanism in ischemic stroke, in this study we established a photo-thrombosis-induced mouse model of focal ischemia and administered freshly isolated mitochondria via the tail vein or to the injury site(in situ). Animal behavior tests, immunofluorescence staining, 2,3,5-triphenyltetrazolium chloride(TTC) staining, mRNA-seq, and western blotting were used to assess mouse anxiety and memory, cortical infarct area, Pyroptosis, and neurogenesis, respectively. Using bioinformatics analysis, western blotting, co-immunoprecipitation, and mass spectroscopy, we identified S100 calcium binding protein A9(S100A9) as a potential regulator of mitochondrial function and determined its possible interacting proteins. Interactions between exogenous and endogenous mitochondria, as well as the effect of exogenous mitochondria on recipient microglia, were assessed in vitro. Our data showed that:(1) mitochondrial transplantation markedly reduced mortality and improved emotional and cognitive function, as well as reducing infarct area, inhibiting Pyroptosis, and promoting cortical neurogenesis;(2) microglial expression of S100A9 was markedly increased by ischemic injury and regulated mitochondrial function;(3) in vitro, exogenous mitochondria enhanced mitochondrial function, reduced redox stress, and regulated microglial polarization and Pyroptosis by fusing with endogenous mitochondria; and(4) S100A9 promoted internalization of exogenous mitochondria by the microglia, thereby amplifying their pro-proliferation and anti-inflammatory effects. Taken togeth

关键词： ischemic stroke microglia mitochondria transplantation neurogenesis Pyroptosis

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Melatonin decreases GSDME mediated mesothelial cell Pyroptosis and prevents peritoneal fibrosis and ultrafiltration failure

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Science China(Life Sciences) 2024年第2期67卷 360-378页

作者： Hongxia Ruan Xuejuan Li Lina Zhou Zihan Zheng Rulin Hua Xu Wang Yuan Wang Yujie Fan Shuwen Guo Lihua Wang Shafiq ur Rahman Ziwei Wang Yuyuan Wei Shuangyan Yu Rongzhi Zhang Qian Cheng Jie Sheng Xue Li Xiaoyan Liu Ruqiang Yuan Xiaoyan Zhang Lihong Chen Guowang Xu Youfei Guan Jing Nie Hongqiang Qin Feng Zheng Advanced Institute for Medical Sciences Dalian Medical UniversityDalian 116044China Department of Nephrology Second HospitalDalian Medical UniversityDalian 116023China CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023China Chongqing International Institute for Immunology Chongqing 401320China Department of Nephrology Shenzhen HospitalSouthern Medical UniversityShenzhen 518101China Wuhu Hospital and Health Science Center East China Normal UniversityShanghai 200241China Peking University First Hospital Peking UniversityBeijing 100034China

Peritoneal fibrosis together with increased capillaries is the primary cause of peritoneal dialysis failure.Mesothelial cell loss is an initiating event for peritoneal fibrosis.We find that the elevated glucose concentrations in peritoneal dialysate drive mesothelial cell Pyroptosis in a manner dependent on caspase-3 and Gasdermin E,driving downstream inflammatory responses,including the activation of macrophages.Moreover,Pyroptosis is associated with elevated vascular endothelial growth factor A and C,two key factors in vascular angiogenesis and lymphatic vessel formation.GSDME deficiency mice are protected from high glucose induced peritoneal fibrosis and ultrafiltration failure.Application of melatonin abrogates mesothelial cell Pyroptosis through a MT1R-mediated action,and successfully reduces peritoneal fibrosis and angiogenesis in an animal model while preserving dialysis efficacy.Mechanistically,melatonin treatment maintains mitochondrial integrity in mesothelial cells,meanwhile activating m TOR signaling through an increase in the glycolysis product dihydroxyacetone phosphate.These effects together with quenching free radicals by melatonin help mesothelial cells maintain a relatively stable internal environment in the face of high-glucose stress.Thus,Melatonin treatment holds some promise in preserving mesothelium integrity and in decreasing angiogenesis to protect peritoneum function in patients undergoing peritoneal dialysis.

关键词： melatonin mitochondria Pyroptosis GSDME peritoneal fibrosis mTOR

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Dual-engineered cartilage-targeting extracellular vesicles derived from mesenchymal stem cells enhance osteoarthritis treatment via miR-223/NLRP3/Pyroptosis axis:Toward a precision therapy

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Bioactive Materials 2023年第12期30卷 169-183页

作者： Weixuan Liu Anqi Liu Xujun Li Ziyang Sun Zhenghua Sun Yaru Liu Gang Wang Dan Huang Hao Xiong Shiyang Yu Xintao Zhang Cunyi Fan Department of Sports Medicine Peking University Shenzhen HospitalShenzhen Peking University-The Hong Kong University of Science and Technology Medical CenterShenzhen518036China Shanghai Engineering Research Center for Orthopedic Material Innovation and Tissue Regeneration Shanghai201306China Department of Orthopedics Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of MedicineShanghai200233China Department of Orthodontics Shanghai Stomatological Hospital&School of StomatologyFudan UniversityShanghai200001China Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases Fudan UniversityShanghai200001China Minhang Hospital Fudan UniversityShanghai201199China

Osteoarthritis(OA)is the most common disabling joint disease with no effective disease modifying drugs.Extracellular vesicles released by several types of mesenchymal stem cells could promote cartilage repair and ameliorate OA pathology in animal models,representing a novel therapeutic strategy.In this study,we demonstrated that extracellular vesicles derived from human umbilical cord mesenchymal stem cells(hUC-EVs)could maintain chondrocyte homeostasis and alleviate OA,and further revealed a novel molecular mechanism of this therapeutic effect.miR-223,which could directly bind with the 3′UTR of NLRP3 mRNA,was found to be a key miRNA for hUC-EVs to exert beneficial effects on inflammation inhibiting and cartilage protecting.For enhancing the effect on mitigating osteoarthritis,exogenous miR-223 was loaded into hUC-EVs by electroporation,and a collagen II-targeting peptide(WYRGRL)was modified onto the surface of hUC-EVs by genetic engineering to achieve a more targeted and efficient RNA delivery to the cartilage.The dual-engineered EVs showed a maximal effect on inhibiting the NLRP3 inflammasome activation and chondrocyte Pyroptosis,and offered excellent results for the treatment of OA.This study provides a novel theoretical basis and a promising therapeutic strategy for the application of engineered extracellular vesicles in OA treatment.

关键词： Osteoarthritis Extracellular vesicles NLRP3 inflammasome Pyroptosis Cartilage-targeting

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The implication of Pyroptosis in cancer immunology:Current advances and prospects

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Genes & Diseases 2023年第6期10卷 2339-2350页

作者： Wei Liu Jinwu Peng Muzhang Xiao Yuan Cai Bi Peng Wenqin Zhang Jianbo Li Fanhua Kang Qianhui Hong Qiuju Liang Yuanliang Yan Zhijie Xu Department of Pathology Xiangya HospitalCentral South UniversityChangshaHunan 410008China Department of Orthopedic Surgery The Second Hospital University of South ChinaHengyangHunan 421001China Department of Pathology Xiangya Changde HospitalChangdeHunan 415000China Department of Burn and Plastic Surgery Xiangya HospitalCentral South UniversityChangshaHunan 410008China Department of Pharmacy Xiangya HospitalCentral South UniversityChangshaHunan 410008China National Clinical Research Center for Geriatric Disorders Xiangya HospitalCentral South UniversityChangshaHunan 410008China

Pyroptosis is a regulated cell death pathway involved in numerous human diseases,especially malignant tumors.Recent studies have identified multiple Pyroptosis-associated signaling molecules,like caspases,gasdermin family and inflammasomes.In addition,increasing in vitro and in vivo studies have shown the significant linkage between Pyroptosis and immune regulation of cancers.Pyroptosis-associated biomarkers regulate the infiltration of tumor immune cells,such as CD4^(+) and CD8^(+) T cells,thus strengthening the sensitivity to therapeutic strategies.In this review,we explained the relationship between Pyroptosis and cancer immunology and focused on the significance of Pyroptosis in immune regulation.We also proposed the future application of Pyroptosis-associated biomarkers in basic research and clinical practices to address malignant behaviors.Exploration of the underlying mechanisms and biological functions of Pyroptosis is critical for immune response and cancer immunotherapy.

关键词： Cancer immunology Immune response Immunotherapy Pyroptosis Inflammasomes

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Protocatechuic acid and quercetin attenuate ETEC-caused IPEC-1 cell inflammation and injury associated with inhibition of necroptosis and Pyroptosis signaling pathways

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Journal of Animal Science and Biotechnology 2023年第4期14卷 1551-1568页

作者： Kan Xiao Mohan Zhou Qingqing Lv Pengwei He Xu Qin Dan Wang Jiangchao Zhao Yulan Liu Hubei Key Laboratory of Animal Nutrition and Feed Science Wuhan Polytechnic UniversityWuhan 430023People’s Republic of China Department of Animal Science Division of AgricultureUniversity of ArkansasFayettevilleAR 72701USA

Background:Necroptosis and Pyroptosis are newly identified forms of programmed cell death,which play a vital role in development of many gastrointestinal disorders.Although plant polyphenols have been reported to protect intestinal health,it is still unclear whether there is a beneficial role of plant polyphenols in modulating necroptosis and Pyroptosis in intestinal porcine epithelial cell line(IPEC-1)infected with enterotoxigenic Escherichia coli(ETEC)K88.This research was conducted to explore whether plant polyphenols including protocatechuic acid(PCA)and quercetin(Que),attenuated inflammation and injury of IPEC-1 caused by ETEC K88 through regulating necroptosis and Pyroptosis signaling pathways.Methods:IPEC-1 cells were treated with PCA(40μmol/L)or Que(10μmol/L)in the presence or absence of ETEC K88.Results:PCA and Que decreased ETEC K88 adhesion and endotoxin level(P<0.05)in cell supernatant.PCA and Que increased cell number(P<0.001)and decreased lactate dehydrogenases(LDH)activity(P<0.05)in cell supernatant after ETEC infection.PCA and Que improved transepithelial electrical resistance(TEER)(P<0.001)and reduced fluorescein isothiocyanate-labeled dextran(FD4)flux(P<0.001),and enhanced membrane protein abundance of occludin,claudin-1 and ZO-1(P<0.05),and rescued distribution of these tight junction proteins(P<0.05)after ETEC infection.PCA and Que also declined cell necrosis ratio(P<0.05).PCA and Que reduced mRNA abundance and concentration of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-8(P<0.001),and down-regulated gene expression of toll-like receptors 4(TLR4)and its downstream signals(P<0.001)after ETEC infection.PCA and Que down-regulated protein abundance of total receptor interacting protein kinase 1(t-RIP1),phosphorylated-RIP1(p-RIP1),p-RIP1/t-RIP1,t-RIP3,p-RIP3,mixed lineage kinase domain-like protein(MLKL),p-MLKL,dynamin-related protein 1(DRP1),phosphoglycerate mutase 5(PGAM5)and high mobility group box 1(HMGB1)(P<0.05)after ETEC infection.Moreover,PCA and Que reduced protein abundance of

关键词： Cell damage ETEC K88 Intestinal inflammation Necroptosis Protocatechuic acid Pyroptosis Quercetin

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Activation-induced Pyroptosis contributes to the loss of MAIT cells in chronic HIV-1 infected patients

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Military Medical Research 2023年第1期10卷 45-63页

作者： Peng Xia Xu-Dong Xing Cui-Xian Yang Xue-Jiao Liao Fu-Hua Liu Hui-Huang Huang Chao Zhang Jin-Wen Song Yan-Mei Jiao Ming Shi Tian-Jun Jiang Chun-Bao Zhou Xi-Cheng Wang Qing He Qing-Lei Zeng Fu-Sheng Wang Ji-Yuan Zhang Senior Department of Infectious Diseases the Fifth Medical Center of Chi‑nese PLA General HospitalNational Clinical Research Center for Infectious DiseasesSavaid Medical SchoolUniversity of Chinese Academy of SciencesBeijing 100039China Department of Infectious Diseases and Hepatology the First Affiliated Hospital of Zhengzhou UniversityZhengzhou 450052China Biomedical Pioneering Innovation Center(BIOPIC) School of Life SciencesPeking UniversityBeijing 100871China Yunnan Infectious Disease Hospital Kunming 650301China The Third People’s Hospital of Shenzhen School of MedicineSouthern University of Science and TechnologyShenz‑hen 518112GuangzhouChina

Background: Mucosal-associated invariant T(MAIT) cells are systemically depleted in human immunodeficiency virus type 1(HIV-1) infected patients and are not replenished even after successful combined antiretroviral therapy(cART).This study aimed to identify the mechanism underlying MAIT cell depletion.Methods: In the present study, we applied flow cytometry, single-cell RNA sequencing and immunohistochemical staining to evaluate the characteristics of pyroptotic MAIT cells in a total of 127 HIV-1 infected individuals, including 69 treatment-naive patients, 28 complete responders, 15 immunological non-responders, and 15 elite controllers, at the Fifth Medical Center of Chinese PLA General Hospital, Beijing, China.Results: Single-cell transcriptomic profiles revealed that circulating MAIT cells from HIV-1 infected subjects were highly activated, with upregulation of Pyroptosis-related genes. Further analysis revealed that increased frequencies of pyroptotic MAIT cells correlated with markers of systemic T-cell activation, microbial translocation, and intestinal damage in cART-naive patients and poor CD4+ T-cell recovery in long-term cART patients. Immunohistochemical staining revealed that MAIT cells in the gut mucosa of HIV-1 infected patients exhibited a strong active gasdermin-D(GSDMD, marker of Pyroptosis) signal near the cavity side, suggesting that these MAIT cells underwent active Pyroptosis in the colorectal mucosa. Increased levels of the proinflammatory cytokines interleukin-12(IL-12) and IL-18 were observed in HIV-1 infected patients. In addition, activated MAIT cells exhibited an increased pyroptotic phenotype after being triggered by HIV-1 virions, T-cell receptor signals, IL-12 plus IL-18, and combinations of these factors, in vitro.Conclusions: Activation-induced MAIT cell Pyroptosis contributes to the loss of MAIT cells in HIV-1 infected patients,which could potentiate disease progression and poor immune reconstitution.

关键词： Acquired immune deficiency syndrome Human immunodeficiency virus Mucosal-associated invariant T cells Pyroptosis Immune reconstitution

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Activation of Pyroptosis by specific organelle-targeting photodynamic therapy to amplify immunogenic cell death for anti-tumor immunotherapy

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Bioactive Materials 2023年第7期25卷 580-593页

作者： Shuang Zeng Chen Chen Liuwei Zhang Xiaosheng Liu Ming Qian Hongyan Cui Jingyun Wang Qixian Chen Xiaojun Peng School of Bioengineering Dalian University of Technology2 Linggong RoadHi-tech ZoneDalian116024China State Key Laboratory of Fine Chemicals Dalian University of Technology2 Linggong RoadHi-tech ZoneDalian116024China

Pyroptosis,a unique lytic programmed cell death,inspired tempting implications as potent anti-tumor strategy in pertinent to its potentials in stimulating anti-tumor immunity for eradication of primary tumors and metastasis.Nonetheless,rare therapeutics have been reported to successfully stimulate Pyroptosis.In view of the intimate participation of reactive oxygen species(ROS)in stimulating Pyroptosis,we attempted to devise a spectrum of well-defined subcellular organelle(including mitochondria,lysosomes and endoplasmic reticulum)-targeting photosensitizers with the aim of precisely localizing ROS(produced from photosensitizers)at the subcellular compartments and explore their potentials in urging Pyroptosis and immunogenic cell death(ICD).The subsequent investigations revealed varied degrees of Pyroptosis upon photodynamic therapy(PDT)towards cancerous cells,as supported by not only observation of the distinctive morphological and mechanistic characteristics of Pyroptosis,but for the first-time explicit validation from comprehensive RNA-Seq analysis.Furthermore,in vivo anti-tumor PDT could exert eradication of the primary tumors,more importantly suppressed the distant tumor and metastatic tumor growth through an abscopal effect,approving the acquirement of specific anti-tumor immunity as a consequence of Pyroptosis.Hence,Pyroptosis was concluded unprecedently by our proposed organelles-targeting PDT strategy and explicitly delineated with molecular insights into its occurrence and the consequent ICD.

关键词： Pyroptosis Tumor Photodynamic therapy Immunogenic cell death Subcellular targeting

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Mesenteric adipose tissue B lymphocytes promote intestinal injury in severe acute pancreatitis by mediating enteric Pyroptosis

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Hepatobiliary & Pancreatic Diseases International 2024年第3期23卷 300-309页

作者： Qing Huang Jia-Wen Liu Hai-Bin Dong Zheng-Jie Wei Jin-Zhe Liu Yu-Tang Ren Xuan Jiang Bo Jiang Department of Gastroenterology Beijing Tsinghua Changgung HospitalSchool of Clinical MedicineTsinghua UniversityBeijing 102218China

Background:Visceral adipose tissue(VAT)has been linked to the severe acute pancreatitis(SAP)prognosis,although the underlying mechanism remains unclear.It has been reported that Pyroptosis worsens SAP.The present study aimed to verify whether mesenteric adipose tissue(MAT,a component of VAT)can cause secondary intestinal injury through the pyroptotic pathway.Methods:Thirty-six male Sprague Dawley(SD)rats were divided into six different groups.Twelve rats were randomly divided into the SAP and control groups.We monitored the changes of MAT and B lymphocytes infiltration in MAT of SAP rats.Twelve SAP rats were injected with MAT B lymphocytes or phosphate buffer solution(PBS).The remaining twelve SAP rats were first injected with MAT B lymphocytes,and then with MCC950(NLRP3 inhibitor)or PBS.We collected blood and tissue samples from pancreas,gut and MAT for analysis.Results:Compared to the control rats,the SAP group showed inflammation in MAT,including higher expression of tumor necrosis factor(TNF-α)and interleukin-6(IL-6),lower expression of IL-10,and histological changes.Flow cytometry analysis revealed B lymphocytes infiltration in MAT but not T lymphocytes and macrophages.The SAP rats also exhibited intestinal injury,characterized by lower expression of zonula occludens-1(ZO-1)and occludin,higher levels of lipopolysaccharide and diamine oxidase,and pathological changes.The expression of NLRP3 and n-GSDMD,which are responsible for Pyroptosis,was increased in the intestine of SAP rats.The injection of MAT B lymphocytes into SAP rats exacerbated the inflammation in MAT.The upregulation of Pyroptosis reduced tight junction in the intestine,which contributed to the SAP progression,including higher inflammatory indicators and worse histological changes.The administration of MCC950 to SAP+MAT B rats downregulated Pyroptosis,which subsequently improved the intestinal barrier and ameliorated inflammatory response of SAP.Conclusions:In SAP,MAT B lymphocytes aggravated local inflammation,and promoted the injury to the int

关键词： Acute pancreatitis Mesenteric adipose tissue B lymphocyte Intestinal barrier Pyroptosis

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H7N9 virus infection triggers lethal cytokine storm by activating gasdermin E-mediated Pyroptosis of lung alveolar epithelial cells

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National Science Review 2022年第1期9卷 47-55页

作者： Xiaopeng Wan Jiqing Li Yupeng Wang Xiaofei Yu Xijun He Jianzhong Shi Guohua Deng Xianying Zeng Guobin Tian Yanbing Li Yongping Jiang Yuntao Guan Chengjun Li Feng Shao Hualan Chen State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences National Institute of Biological Sciences

The H7 N9 influenza virus emerged in China in 2013,causing more than 1560 human infections,39% of which were fatal.A ’cytokine storm’ in the lungs of H7 N9 patients has been linked to a poor prognosis and death;however,the underlying mechanism that triggers the cytokine storm is unknown.Here,we found that efficient replication of the H7 N9 virus in mouse lungs activates gasdermin E(GSDME)-mediated Pyroptosis in alveolar epithelial cells,and that the released cytosolic contents then trigger a cytokine storm.Knockout of Gsdme switched the manner of death of A549 and human primary alveolar epithelial cells from Pyroptosis to apoptosis upon H7 N9 virus infection,and Gsdme knockout mice survived H7 N9 virus lethal infection.Our findings reveal that GSDME activation is a key and unique mechanism for the pulmonary cytokine storm and lethal outcome of H7 N9 virus infection and thus opens a new door for the development of antivirals against the H7 N9 virus.

关键词： H7N9 virus cytokine storm gasdermin E Pyroptosis

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