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Metadherin promotes stem cell phenotypes and correlated with immune infiltration in hepatocellular carcinoma

World Journal of Gastroenterology 2024年第8期30卷 901-918页

作者： Yi-Ying Wang Mei-Mei Shen Jian Gao Department of Gastroenterology The Second Affiliated Hospital of Chongqing Medical UniversityChongqing 400010China Department of Gastroenterology and Hepatology The Second Affiliated Hospital of Chongqing Medical UniversityChongqing 400010China

BACKGROUND Metadherin(MTDH)is a key oncogene in most cancer types,including hepato-cellular carcinoma(HCC).Notably,MTDH does not affect the stemness pheno-type or immune infiltration of HCC.AIM To explore the role of MTDH on stemness and immune infiltration in HCC.METHODS MTDH expression in HCC tissues was detected using TCGA and GEO databases.Immunohistochemistry was used to analyze the tissue samples.MTDH was stably knocked down or overexpressed by lentiviral transfection in the two HCC cell lines.The invasion and migration abilities of HCC cells were evaluated using Matrigel invasion and wound healing assays.Next,we obtained liver cancer stem cells from the spheroids by culturing them in a serum-free medium.Gene expression was determined by western blotting and quantitative reverse transcri-ption PCR.Flow cytometry,immunofluorescence,and tumor sphere formation assays were used to characterize stem-like cells.The effects of MTDH inhibition on tumor growth were evaluated in vivo.The correlation of MTDH with immune cells,immunomodulators,and chemokines was analyzed using ssGSEA and TISIDB databases.RESULTS HCC tissues expressed higher levels of MTDH than normal liver tissues.High MTDH expression was associated with a poor prognosis.HCC cells overex-pressing MTDH exhibited stronger invasion and migration abilities,exhibited a stem cell-like phenotype,and formed spheres;however,MTDH inhibition attenuated these effects.MTDH inhibition suppressed HCC progression and CD133 expression in vivo.MTDH was positively correlated with immature dendritic,T helper 2 cells,central memory CD8^(+)T,memory B,activated dendritic,natural killer(NK)T,NK,activated CD4^(+)T,and central memory CD4^(+)T cells.MTDH was negatively correlated with activated CD8^(+)T cells,eosinophils,activated B cells,monocytes,macrophages,and mast cells.A positive correlation was observed between the MTDH level and CXCL2 expression,whereas a negative correlation was observed between the MTDH level and CX3CL1 and CXCL12 expression.CONCLUSION High levels of M

关键词： Metadherin Hepatocellular carcinoma Cancer stem cells Immune infiltration

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Integrated neural tracing and in-situ barcoded sequencing reveals the logic of SCN efferent circuits in regulating circadian behaviors

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Science China(Life Sciences) 2024年第3期67卷 518-528页

作者： Meimei Liao Xinwei Gao Chen Chen Qi Li Qingchun Guo He Huang Erquan Zhang Dapeng Ju College of Biological Sciences China Agriculture UniversityBeijing 100193China National Institute of Biological Sciences(NIBS) Beijing 102206China Chinese Institute for Brain Research Beijing 102206China School of Biomedical Engineering Capital Medical UniversityBeijing 100069China Department of Anesthesiology The Second Affiliated HospitalChongqing Medical UniversityChongqing 401336China Tsinghua Institute of Multidisciplinary Biomedical Research Tsinghua UniversityBeijing 102206China

The circadian clock coordinates rhythms in numerous physiological processes to maintain organismal homeostasis. Since the suprachiasmatic nucleus(SCN) is widely accepted as the circadian pacemaker, it is critical to understand the neural mechanisms by which rhythmic information is transferred from the SCN to peripheral clocks. Here, we present the first comprehensive map of SCN efferent connections and suggest a molecular logic underlying these projections. The SCN projects broadly to most major regions of the brain, rather than solely to the hypothalamus and thalamus. The efferent projections from different subtypes of SCN neurons vary in distance and intensity, and blocking synaptic transmission of these circuits affects circadian rhythms in locomotion and feeding to different extents. We also developed a barcoding system to integrate retrograde tracing with in-situ sequencing, allowing us to link circuit anatomy and spatial patterns of gene expression. Analyses using this system revealed that brain regions functioning downstream of the SCN receive input from multiple neuropeptidergic cell types within the SCN, and that individual SCN neurons generally project to a single downstream brain region.This map of SCN efferent connections provides a critical foundation for future investigations into the neural circuits underlying SCNmediated rhythms in physiology. Further, our new barcoded tracing method provides a tool for revealing the molecular logic of neuronal circuits within heterogeneous brain regions.

关键词： circadian rhythms SCN output circuit neural tracing barcoded GFP in situ sequencing

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PCK1 attenuates tumor stemness via activating the Hippo signaling pathway in hepatocellular carcinoma

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Genes & Diseases 2024年第4期11卷 340-352页

作者： Rui Liu Yi Liu Wenlu Zhang Guiji Zhang Zhirong Zhang Luyi Huang Ni Tang Kai Wang Key Laboratory of Molecular Biology for Infectious Diseases(Ministry of Education) Institute for Viral HepatitisDepartment of Infectious DiseasesThe Second Affiliated HospitalChongqing Medical UniversityChongqing 400016China Department of Clinical Laboratory Sichuan Clinical Research Center for CancerSichuan Cancer Hospital&InstituteSichuan Cancer CenterAffiliated Cancer Hospital of University of Electronic Science and Technology of ChinaChengduSichuan 610041China

Liver cancer stem cells were found to rely on glycolysis as the preferred metabolic program.Phosphoenolpyruvate carboxylase 1(PCK1),a gluconeogenic metabolic enzyme,is down-regulated in hepatocellular carcinoma and is closely related to poor prognosis.The onco-genesis and progression of tumors are closely related to cancer stem cells.It is not completely clear whether the PCK1 deficiency increases the stemness of hepatoma cells and promotes the oncogenesis of hepatocellular carcinoma.Herein,the results showed that PCK1 inhibited the self-renewal property of hepatoma cells,reduced the mRNA level of cancer stem cell markers,and inhibited tumorigenesis.Moreover,PCK1 increased the sensitivity of hepatocellular carci-noma cells to sorafenib.Furthermore,we found that PCK1 activated the Hippo pathway by enhancing the phosphorylation of YAP and inhibiting its nuclear translocation.Verteporfin reduced the stemness of hepatoma cells and promoted the pro-apoptotic effect of sorafenib.Thus,combined treatment with verteporfin and sorafenib may be a potential anti-tumor strat-egy in hepatocellular carcinoma.

关键词： Cancer stem cells Chemoresistance Gluconeogenesis Hepatocellular carcinoma YAP

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Azvudine reduces the in-hospital mortality of COVID-19 patients:A retrospective cohort study

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Acta Pharmaceutica Sinica B 2023年第11期13卷 4655-4660页

作者： Kaican Zong Hui Zhou Wen Li E Jiang Yi Liu Shiying Li Department of Respiratory Medicine the Seventh People's Hospital of Chongqing(Affiliated Central Hospital of Chongqing University of Technology)Chongqing 400054China Department of Infectious Diseases Key Laboratory of Molecular Biology for Infectious Diseases(Ministry of Education)Institute for Viral Hepatitisthe Second Affiliated HospitalChongqing Medical UniversityChongqing 400010China

In our retrospective cohort study,we aim to explore whether Azvudine modifies the risk of death in COVID-19 patients.It was conducted on the medical records of patients,consecutively admitted for COVID-19 pneumonia to two hospitals in chongqing,China.Based on Azvudine treatment exposure,the patients were divided into Azvudine group and non-Azvudine group.We used 1:2 ratio propensity score matching(PSM)in our study to adjust for confounding factors and differences between Azvudine and non-Azvudine groups.There were 1072 patients included in our original cohort.With 1:2 ratio PSM,the Azvudine group included 195 patients and non-Azvudine group included 390 patients.The results showed that Azvudine treatment was associated with improved in-hospital mortality in overall population(OR 0.375,95%CI 0.225-0.623,Phospital mortality in overall population(11%vs.24%,Phospital mortality in overall COVID-19 patients,severe,and critical subgroup population.

关键词： Azvudine COVID-19 Pneumonia Mortality

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Single-cell and spatial heterogeneity landscapes of mature epicardial cells

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药物分析学报:英文版 2023年第8期13卷 894-907页

作者： Jianlin Du Xin Yuan Haijun Deng Rongzhong Huang Bin Liu Tianhua Xiong Xianglin Long Ling Zhang Yingrui Li Qiang She Department of Cardiology The Second Affiliated Hospital of Chongqing Medical UniversityChongqing400010China Department of Nephrology The Second Affiliated Hospital of Chongqing Medical UniversityChongqing400010China Key Laboratory of Molecular Biology for Infectious Diseases Ministry of EducationInstitute for Viral HepatitisThe Second Affiliated Hospital of Chongqing Medical UniversityChongqing400010China Precision Medicine Center The Second Affiliated Hospital of Chongqing Medical UniversityChongqing400010China Basic Medicine Research and Innovation Center for Novel Target and Therapeutic Intervention Ministry of EducationThe Second Affiliated Hospital of Chongqing Medical UniversityChongqing400010China

Tbx18,Wt1,and Tcf21 have been identified as epicardial markers during the early embryonic stage.However,the gene markers of mature epicardial cells remain unclear.Single-cell transcriptomic analysis was performed with the Seurat,Monocle,and CellphoneDB packages in R software with standard procedures.Spatial transcriptomics was performed on chilled Visium Tissue Optimization Slides(10x Genomics)and Visium Spatial Gene Expression Slides(10x Genomics).Spatial transcriptomics analysis was performed with Space Ranger software and R software.Immunofluorescence,whole-mount RNA in situ hybridization and X-gal staining were performed to validate the analysis results.Spatial transcriptomics analysis revealed distinct transcriptional profiles and functions between epicardial tissue and non-epicardial tissue.Several gene markers specific to postnatal epicardial tissue were identified,including Msln,C3,Efemp1,and Upk3b.Single-cell transcriptomic analysis revealed that cardiac cells from wildtype mouse hearts(from embryonic day 9.5 to postnatal day 9)could be categorized into six major cell types,which included epicardial cells.Throughout epicardial development,Wt1,Tbx18,and Upk3b were consistently expressed,whereas genes including Msln,C3,and Efemp1 exhibited increased expression during the mature stages of development.Pseudotime analysis further revealed two epicardial cell fates during maturation.Moreover,Upk3b,Msln,Efemp1,and C3 positive epicardial cells were enriched in extracellular matrix signaling.Our results suggested Upk3b,Efemp1,Msln,C3,and other genes were mature epicardium markers.Extracellular matrix signaling was found to play a critical role in the mature epicardium,thus suggesting potential therapeutic targets for heart regeneration in future clinical practice.

关键词： Epicardial cells Gene markers Single-cell sequencing Spatial transcriptomics

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Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis

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Military medical Research 2022年第6期9卷 668-685页

作者： Xue Zeng Yun-Dong Zhang Rui-Yan Ma Yuan-Jing Chen Xin-Ming Xiang Dong-Yao Hou Xue-Han Li He Huang Tao Li Chen-Yang Duan Department of Anaesthesiology the Second Affiliated Hospital of Chongqing Medical UniversityChongqing 400010China Department of Neurology the Third Affliated Hospital of Chongqing Medical UniversityChongqing 401120China Department of Cardiovascular Surgery Xinqiao HospitalArmy Medical UniversityChongqing 400037China State Key Laboratory of Trauma Burns and Combined InjuryDepartment of Shock and TransfusionDaping HospitalArmy Medical UniversityChongqing 400042China

Background: Cerebral ischemia-reperfusion injury(CIRI) refers to a secondary brain injury that can occur when the blood supply to the ischemic brain tissue is restored. However, the mechanism underlying such injury remains elusive.Methods: The 150 male C57 mice underwent middle cerebral artery occlusion(MCAO) for 1 h and reperfusion for 24 h,Among them, 50 MCAO mice were further treated with Mitochondrial division inhibitor 1(Mdivi-1) and 50 MCAO mice were further treated with N-acetylcysteine(NAC). SH-SY5Y cells were cultured in a low-glucose culture medium for 4 h under hypoxic conditions and then transferred to normal conditions for 12 h. Then, cerebral blood flow, mitochondrial structure, mitochondrial DNA(mtDNA) copy number, intracellular and mitochondrial reactive oxygen species(ROS),autophagic flux, aggresome and exosome expression profiles, cardiac tissue structure, mitochondrial length and cristae density, mtDNA and ROS content, as well as the expression of Drp1-Ser616/Drp1, RIP1/RIP3, LC3 II/I, TNF-α,IL-1β, etc., were detected under normal or Drp1 interference conditions.Results: The mtDNA content, ROS levels, and Drp1-Ser616/Drp1 were elevated by 2.2, 1.7 and 2.7 times after CIRI(P0.05), it increased the packaging and extracellular secretion of exosomes containing p62 by 4–5 times, which could be decreased by Mdivi-1, Drp1 shRNA, and Nec-1(P<0.05). Furthermore, TNF-α and IL-1βincreased in CIRI-derived exosomes could increase RIP3 phosphoryla

关键词： Cerebral ischemia-reperfusion(CIRI) Oxygen-glucose deprivation/reoxygenation(OGD/R) Drp1 p62 LC3 II/I Reactive oxygen species(ROS) RIP1/RIP3 Autophagy Exosome Inflammatory

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Elevated neutrophil extracellular traps by HBV-mediated S100A9-TLR4/RAGE-ROS cascade facilitate the growth and metastasis of hepatocellular carcinoma

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Cancer Communications 2023年第2期43卷 225-245页

作者： Xi Zhan RuiWu Xue-Hua Kong Yan You Kun He Xiao-Yu Sun Yong Huang Wei-Xian Chen Liang Duan Department of Laboratory Medicine The Second Affiliated Hospital of Chongqing Medical UniversityChongqing 400000P.R.China Department of Laboratory Medicine The First Affiliated Hospital of Chongqing Medical UniversityChongqing 400016P.R.China Key Laboratory of Laboratory medical Diagnostics Ministry of EducationDepartment of Laboratory MedicineChongqing Medical UniversityChongqing 400010P.R.China Department of Pathology The Second Affiliated Hospital of Chongqing Medical UniversityChongqing 400000P.R.China Department of Hepatobiliary Surgery The Second Affiliated Hospital of Chongqing Medical UniversityChongqing 400000P.R.China

Background:Neutrophil extracellular traps(NETs)are considered significant contributors to cancer progression,especially metastasis.However,it is still unclear whether NETs are involved in hepatitis B virus(HBV)-related hepatocarcinogenesis and have potential clinical significance during evaluation and management for hepatocellular carcinoma(HCC).In this study,we aimed to investigate the functional mechanism of NETs in HBV-related hepatocarcinogenesis and their clinical significance.Methods:A total of 175 HCC patients with and without HBV infection and 58 healthy controls were enrolled in this study.NETs weremeasured in tissue specimens,freshly isolated neutrophils and blood serum from these patients,and the correlation of circulating serum NETs levels with malignancy was evaluated.The mechanism by which HBV modulates NETs formation was explored using cell-based studies.In addition,in vitro and in vivo experiments were further performed to clarify the functional mechanism of NETs on the growth and metastasis of HCC.Results:We observed an elevated level of NETs in blood serum and tissue specimens from HCC patients,especially those infected with HBV.NETs facilitated the growth and metastasis of HCC both in vitro and in vivo,which were mainly dominated by increased angiogenesis,epithelial-mesenchymal transition(EMT)-related cell migration,matrix metalloproteinases(MMPs)-induced extracellular matrix(ECM)degradation and NETs-mediated cell trapping.Inhibition of NETs generation by DNase 1 effectively abrogated the NETs-aroused HCC growth and metastasis.In addition,HBV-induced S100A9 accelerated the generation of NETs,which was mediated by activation of toll-like receptor(TLR4)/receptor for advanced glycation end products(RAGE)-reactive oxygen species(ROS)signaling.Further,circulatory NETs were found to correlate with viral load,TNM stage and metastasis status in HBV-related HCC,and the identified NETs could predict extrahepatic metastasis,with an area under the ROC curve(AUC)of 0.83 and 90.3%sensitivity and 62.8%specifi

关键词： hepatocellular carcinoma hepatitis B virus metastasis neutrophil extracellular trap RAGE ROS S100A9 TLR4

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Laparoscopic liver resection for hepatocellular carcinoma complicated with significant portal hypertension:A propensity score-matched survival analysis

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Hepatobiliary & Pancreatic Diseases International 2023年第4期22卷 358-365页

作者： Zhang-You Guo Yuan Hong Bing Tu Yao Cheng Xiao-Mei Wang Department of Minimally Invasive Interventional Therapy Yunnan Cancer HospitalThe Third Affiliated Hospital of Kunming Medical UniversityYunnan Cancer CenterKunming 650118China medical Laboratory The First Affiliated Hospital of Kunming Medical UniversityInstitute of Experimental Diagnostics of Yunnan ProvinceKey Laboratory of Laboratory Medicine of Yunnan ProvinceKunming 650032China Department of Hepatobiliary Surgery The Second Affiliated HospitalChongqing Medical UniversityChongqing 400010China

Background: Significant portal hypertension(SPH) is a relative contraindication for patients with resectable hepatocellular carcinoma(HCC). However, increasing evidence indicates that liver resection is feasible for HCC patients with SPH. Methods: HCC patients with cirrhosis who underwent laparoscopic liver resection(LLR) in two centers from January 2013 to April 2018 were included. Surgical and survival outcomes were analyzed to explore potential prognostic factors. Propensity score matching(PSM) analysis was performed to minimize bias. Results: A total of 165 patients were divided into two groups based on the presence(SPH, n = 76) or absence(non-SPH, n = 89) of SPH. Patients in the SPH group had longer operative time, more blood loss, and more advanced TNM stage than patients in the non-SPH group( P hospitalization(16 vs. 15 days, P = 0.203) between the SPH and non-SPH groups before PSM. Similar results were obtained after PSM. The 1-, 3-, and 5-year overall survival(OS) and recurrence-free survival rates in the SPH group were not significantly different from those in the non-SPH group both before and after PSM(log-rank P > 0.05). After PSM, alpha-fetoprotein(AFP) ≥ 400 μg/L [hazard ratio(HR) = 4.71, 95% confidence interval(CI): 2.69-8.25], ascites(HR = 2.18, 95% CI: 1.30-3.66), American Society of Anesthesiologists(ASA) classification(Ⅲ vs. Ⅱ)(HR = 2.13, 95% CI: 1.11-4.07) and tumor diameter > 5 cm(HR = 3.91, 95% CI: 2.02-7.56) independently predicted worse OS. Conclusions: LLR for patients with HCC complicated with SPH appears feasible at the price of increasing operative time and blood loss. AFP, ascites, ASA classification and tumor diameter may predict the prognosis of HCC complicated with SPH after LLR.

关键词： Hepatocellular carcinoma Significant portal hypertension Laparoscopic liver resection Propensity score matching analysis

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LIFU-responsive nanomedicine enables acoustic droplet vaporization-induced apoptosis of macrophages for stabilizing vulnerable atherosclerotic plaques

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Bioactive Materials 2022年第10期7卷 120-133页

作者： Jingxin Hou Jun Zhou Meiqi Chang Guangcheng Bao Jie Xu Man Ye Yixin Zhong Shuling Liu Junrui Wang Wei Zhang Haitao Ran Zhigang Wang Yu Chen Dajing Guo Department of Radiology Second Affiliated Hospital of Chongqing Medical UniversityChongqing400010PR China Materdicine Lab School of Life SciencesShanghai UniversityShanghai200444PR China chongqing Key Laboratory of Ultrasound Molecular Imaging&Department of Ultrasound Second Affiliated Hospital of Chongqing Medical UniversityChongqing400010PR China State Key Laboratory of High Performance Ceramics and Superfine Microstructure Shanghai Institute of CeramicsChinese Academy of SciencesShanghai200050PR China Department of Radiology Children’s Hospital of Chongqing Medical UniversityChongqing400014PR China

Due to the high risk of tearing and rupture,vulnerable atherosclerotic plaques would induce serious cardiovascular and cerebrovascular diseases.Despite the available clinical methods can evaluate the vulnerability of plaques and specifically treat vulnerable plaques before a cardiovascular event,but the efficiency is still low and undesirable.Herein,we rationally design and engineer the low-intensity focused ultrasound(LIFU)-responsive FPD@CD nanomedicine for the highly efficient treatment of vulnerable plaques by facilely loading phase transition agent perfluorohexane(PFH)into biocompatible PLGA-PEG-PLGA nanoparticles(PPP NPs)and then attaching dextran sulphate(DS)onto the surface of PPP NPs for targeting delivery.DS,as a typical macrophages-targeted molecule,can achieve the precise vaporization of NPs and subsequently controllable apoptosis of RAW 264.7 macrophages as induced by acoustic droplet vaporization(ADV)effect.In addition,the introduction of DiR and Fe3O4 endows nanomedicine with near-infrared fluorescence(NIRF)and magnetic resonance(MR)imaging capabilities.The engineered FPD@CD nanomedicine that uses macrophages as therapeutic targets achieve the conspicuous therapeutic effect of shrinking vulnerable plaques based on in vivo and in vitro evaluation outcomes.A reduction of 49.4%of vascular stenosis degree in gross pathology specimens were achieved throughout the treatment period.This specific,efficient and biosafe treatment modality potentiates the biomedical application in patients with cardiovascular and cerebrovascular diseases based on the relief of the plaque rupture concerns.

关键词： Vulnerable plaques Acoustic droplet vaporization Macrophages Apoptosis Nanomedicine

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PCK1 dysregulation in cancer: Metabolic reprogramming, oncogenic activation, and therapeutic opportunities

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Genes & Diseases 2023年第1期10卷 101-112页

作者： Jin Xiang Kai Wang Ni Tang Key Laboratory of Molecular Biology for Infectious Diseases(Ministry of Education) Institute for Viral HepatitisDepartment of Infectious DiseasesThe Second Affiliated Hospital of Chongqing Medical UniversityChongqing 400010China

The last few decades have witnessed an advancement in our understanding of multiple cancer cell pathways related to metabolic reprogramming.One of the most important cancer hallmarks,including aerobic glycolysis(the Warburg effect),the central carbon pathway,and multiple-branch metabolic pathway remodeling,enables tumor growth,progression,and metastasis.Phosphoenolpyruvate carboxykinase1(PCK1),a key rate-limiting enzyme in gluconeogenesis,catalyzes the conversion of oxaloacetate to phosphoenolpyruvate.PCK1 expression in gluconeogenic tissues is tightly regulated during fasting.In tumor cells,PCK1 is regulated in a cell-autonomous manner rather than by hormones or nutrients in the extracellular environment.Interestingly,PCK1has ananti-oncogenic role in gluconeogenic organs(the liver and kidneys),but a tumor-promoting role in cancers arising from non-gluconeogenic organs.Recent studies have revealed that PCK1 has metabolic and non-metabolic roles in multiple signaling networks linking metabolic and oncogenic pathways.Aberrant PCK1 expression results in the activation of oncogenic pathways,accompanied by metabolic reprogramming,to maintain tumorigenesis.In this review,we summarize the mechanisms underlying PCK1 expression and regulation,and clarify the crosstalk between aberrant PCK1 expression,metabolic rewiring,and signaling pathway activation.In addition,we highlight the clinical relevance of PCK1 and its value as a putative cancer therapeutic target.

关键词： Gluconeogenesis Metabolism Oncogenesis PCK1 Tumor

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