Inflammatory/immune-suppressive microenvironment characteristics of pancreatic cancer patients with Shi-Re syndrome identified by extracellular vesicle long RNA profiling

Background:Numerous long RNAs were detected in extracellular vesicles(EVs),some of which were related with the tissue origins and immune cell types.This study examined the molecular basis of different traditional Chinese medicine syndrome diagnoses(also called syndrome differentiation)in pancreatic ductal adenocarcinoma.Methods:128 pancreatic ductal adenocarcinoma patients with different syndrome diagnoses were retrospectively reviewed in this study.Long RNA sequencing was conducted to analyze the EV long RNA profile of plasma samples.Differentially regulated EV long RNAs were annotated and assessed for Gene Ontology pathway enrichment using DAVID.The online program xCell were used to perform the cell-type enrichment analysis.Results:An average of 15,000 annotated genes,mainly including messenger RNAs,were stably detected per sample.Different syndrome diagnoses exhibited unique EV mRNA expression profiles and therefore different enriched pathways.Gene Set Enrichment Analysis discovered transforming growth factor-βand kirsten rat sarcoma viral oncogene homolog signaling activation as the hallmarks of cancer with Shi-Re syndrome.Cell-type enrichment analysis also revealed a varied inflammation/immune cell type distribution among patients with or without Shi-Re diagnosis.Mast cells,platelets and Tregs were significantly enriched but basophils,common lymphoid progenitors,dendritic cells,and conventional dendritic cells were decreased in patients with Shi-Re diagnosis compared with patients without Shi-Re diagnosis.Conclusion:We identified the hallmarks of cancer with different syndrome diagnoses based on plasma EV long RNA sequencing.In particular,transforming growth factor-βand kirsten rat sarcoma viral oncogene homolog signaling activation were the hallmarks of Shi-Re syndrome,which contribute to shape an inflammatory/immune-suppressive tumor microenvironment in pancreatic ductal adenocarcinoma.

Decoding the complexity of metastasis

Cancer metastasis remains one of the most confounding questions in oncology1,2.Although current cutting-edge techniques enable very early detection of tumors,profiling whether a tumor has already begun to spread and where it has attempted to colonize remains a major hurdle.Indeed,metastatic seeding events exhibit remarkable temporal and spatial heterogeneity,wherein the origin(primary site)and destination(metastatic site)are highly dynamic.For example,liver metastasis is particularly common and remains a leading cause of mortality3.

Bispecific antibodies targeting immunomodulatory checkpoints for cancer therapy

Advances in antibody engineering have led to the generation of more innovative antibody drugs,such as bispecific antibodies(bs Abs).Following the success associated with blinatumomab,bs Abs have attracted enormous interest in the field of cancer immunotherapy.By specifically targeting two different antigens,bs Abs reduce the distance between tumor and immune cells,thereby enhancing tumor killing directly.There are several mechanisms of action upon which bs Abs have been exploited.Accumulating experience on checkpoint-based therapy has promoted the clinical transformation of bs Abs targeting immunomodulatory checkpoints.Cadonilimab(PD-1×CTLA-4)is the first approved bs Ab targeting dual inhibitory checkpoints,which confirms the feasibility of bs Abs in immunotherapy.In this review we analyzed the mechanisms by which bs Abs targeting immunomodulatory checkpoints and their emerging applications in cancer immunotherapy.

Epigenome-wide development and validation of a prognostic methylation score in intrahepatic cholangiocarcinoma based on machine learning strategies

Background:Clinical parameter-based nomograms and staging systems provide limited information for the prediction of survival in intrahepatic cholangiocarcinoma(ICC)patients.In this study,we developed a methylation signature that precisely predicts overall survival(OS)after surgery.Methods:An epigenome-wide study of DNA methylation based on whole-genome bisulfite sequencing(WGBS)was conducted for two independent cohorts(discovery cohort,n=164;validation cohort,n=170)from three hepatobiliary centers in China.By referring to differentially methylated regions(DMRs),we proposed the concept of prognostically methylated regions(PMRs),which were composed of consecutive prognostically methylated CpGs(PMCs).Using machine learning strategies(Random Forest and the least absolute shrinkage and selector regression),a prognostic methylation score(PMS)was constructed based on 14 PMRs in the discovery cohort and confirmed in the validation cohort.Results:The C-indices of the PMS for predicting OS in the discovery and validation cohorts were 0.79 and 0.74,respectively.In the whole cohort,the PMS was an independent predictor of OS[hazard ratio(HR)=8.12;95% confidence interval(CI):5.48-12.04;P<0.001],and the C-index(0.78)of the PMS was significantly higher than that of the Johns Hopkins University School of Medicine(JHUSM)nomogram(0.69,P<0.001),the Eastern Hepatobiliary Surgery Hospital(EHBSH)nomogram(0.67,P<0.001),American Joint Committee on Cancer(AJCC)tumor-node-metastasis(TNM)staging system(0.61,P<0.001),and MEGNA prognostic score(0.60,P<0.001).The patients in quartile 4 of PMS could benefit from adjuvant therapy(AT)(HR=0.54;95%CI:0.32-0.91;log-rank P=0.043),whereas those in the quartiles 1-3 could not.However,other nomograms and staging system failed to do so.Further analyses of potential mechanisms showed that the PMS was associated with tumor biological behaviors,pathway activation,and immune microenvironment.Conclusions:The PMS could improve the prognostic accuracy and identify patients who would benefit from AT for ICC patients,and might facilitate decisions in treatment of ICC patients.

STING signaling activation inhibits HBV replication and attenuates the severity of liver injury and HBV-induced fibrosis

The covalently closed circular DNA(cccDNA)of HBV plays a crucial role in viral persistence and is also a risk factor for developing HBV-induced diseases,including liver fibrosis.Stimulator of interferon genes(STING),a master regulator of DNA-mediated innate immune activation,is a potential therapeutic target for viral infection and virus-related diseases.In this study,agonist-induced STING signaling activation in macrophages was revealed to inhibit cccDNA-mediated transcription and HBV replication via epigenetic modification in hepatocytes.Notably,STING activation could efficiently attenuate the severity of liver injury and fibrosis in a chronic recombinant cccDNA(rcccDNA)mouse model,which is a proven suitable research platform for HBV-induced fibrosis.Mechanistically,STING-activated autophagic flux could suppress macrophage inflammasome activation,leading to the amelioration of liver injury and HBV-induced fibrosis.Overall,the activation of STING signaling could inhibit HBV replication through epigenetic suppression of cccDNA and alleviate HBV-induced liver fibrosis through the suppression of macrophage inflammasome activation by activating autophagic flux in a chronic HBV mouse model.This study suggests that targeting the STING signaling pathway may be an important therapeutic strategy to protect against persistent HBV replication and HBV-induced fibrosis.

USP47-mediated deubiquitination and stabilization of YAP contributes to the progression of colorectal cancer

Dear Editor,The Hippo tumor suppressor pathway plays an important role in development and tumorigenesis.Yes-associated protein(YAP)is a major effector of the Hippo pathway regulating the expression of genes involved in cell proliferation,cell death,and cell differentiation(Pan,2010;Johnson and Haider,2014;Piccolo etal.,2014;Yu etal.,2015).In intestine,YAP is mainly expressed in leucine rich repeat containing G protein-coupled receptor 5(LGR5)marked stem cells,and is required for regeneration following tissue damage and the development of colon cancer in mice(Cai etal.,2010;Barry etal.,2013;Wang etal.,2017).Moreover,in colorectal cancer(CRC)specimens,elevated YAP expression and activity is frequently observed(Steinhardt et al.,2008).It has been shown that high YAP expression is associated with cancer metastasis,poor prognosis,resistance to chemotherapy,and greater possibility of relapse(Johnson and Haider,2014;Yu etal.,2015).

The influence of ^(18)F-fluorodeoxyglucose positron emission tomography/computed tomography on the N-and M-staging and subsequent clinical management of intrahepatic cholangiocarcinoma

Background:Intrahepatic cholangiocarcinoma(ICC)is a highly metastatic cancer.^(18)F-fluorodeoxyglucose positron emission tomography/computed tomography(^(18)F-FDG PET/CT)enables sensitive tumor and metastasis detection.Our aim is to evaluate the influence of pre-treatment PET/CT on the N-and M-staging and subsequent clinical management in ICC patients.Methods:Between August 2010 and August 2018,660 consecutive ICC patients,without prior anti-tumor treatments nor other malignancies,were enrolled.The diagnostic performance of PET/CT on the N-and M-staging was compared with conventional imaging,and the preoperative staging accuracy and treatment re-allocation by PET/CT were retrospectively calculated.Survival difference was compared between patients receiving PET/CT or not after propensity score matching.Results:Patients were divided into group A(n=291)and group B(n=369)according to whether PET/CT was performed.Among 291 patients with both PET/CT and conventional imaging for staging in group A,PET/CT showed significantly higher sensitivity(83.0%vs.70.5%,P=0.001),specificity(88.3%vs.74.9%,P<0.001)and accuracy(86.3%vs.73.2%,P<0.001)than conventional imaging in diagnosing regional lymph node metastasis,as well as higher sensitivity(87.8%vs.67.6%,P<0.001)and accuracy(93.5%vs.89.3%,P=0.023)in diagnosing distant metastasis.Overall,PET/CT improved the accuracy of preoperative staging from 60.1%to 71.8%(P<0.001),and modified clinical treatment strategy in 5.8%(17/291)of ICC patients,with unique roles in different tumor-node-metastasis(TNM)stages.High tumor-to-non-tumor ratio(TNR)predicted poor overall survival[hazard ratio(HR)=2.17;95%confidence interval(CI):1.49-3.15;P<0.001].Furthermore,patients performing PET/CT had longer overall survival compared with those without PET/CT(HR=0.74;95%CI:0.58-0.93;P=0.011)after propensity score matching.Conclusions:PET/CT was valuable for diagnosing regional lymph node metastasis and distant metastasis in ICC patients,and facilitated accurate tumor staging and optimal treatment allocation.

Unraveling the Emerging Niche Role of Hepatic Stellate Cell-derived Exosomes in Liver Diseases

Hepatic stellate cells(HSCs)play an essential role in various liver diseases,and exosomes are critical mediators of intercellular communication in local and distant microenvironments.Cellular crosstalk between HSCs and surrounding multiple tissue-resident cells promotes or inhibits the activation of HSCs.Substantial evidence has revealed that HSC-derived exosomes are involved in the occurrence and development of liver diseases through the regulation of retinoid metabolism,lipid metabolism,glucose metabolism,protein metabolism,and mitochondrial metabolism.HSCderived exosomes are underpinned by vehicle molecules,such as mRNAs and microRNAs,that function in,and significantly affect,the processes of various liver diseases,such as acute liver injury,alcoholic liver disease,nonalcoholic fatty liver disease,viral hepatitis,fibrosis,and cancer.As such,numerous exosomes derived from HSCs or HSCassociated exosomes have attracted attention because of their biological roles and translational applications as potential targets for therapeutic targets.Herein,we review the pathophysiological and metabolic processes associated with HSC-derived exosomes,their roles in various liver diseases and their potential clinical application.

Frequent RNF43 mutation contributes to moderate activation of Wnt signaling in colorectal signet-ring cell carcinoma

Dear Editor,Signet-ring cell carcinoma(SRCC)is a rare subtype of colorectal cancer(CRC)characterized histologically by the accumulation of mucins in the cytoplasm and displacement of nuclei to the cellular periphery,accounting for about 1%CRC(Fig.S1A)(Borger et al.,2007).Compare to common subtypes of CRC,such as adenocarcinoma(AC)and mucinous adenocarcinoma(MAC),SRCC is associated with aggressive behaviors and younger age at presentation(Kang et al.,2005;Sung et al.,2008;Nitsche et al.,2013;Hugen et al.,2014;Inamura et al.,2015).A retrospective analysis of CRC patient's data at Fudan University Shanghai Cancer Center(FUSCC)also indicated a worse overall and disease-free survival of SRCC patients(Fig.S1B and S1C,Table S1).

Oxidative stress-initiated one-carbon metabolism drives the generation of interleukin-10-producing B cells to resolve pneumonia

The metabolic reprogramming underlying the generation of regulatory B cells during infectious diseases remains unknown.Using a Pseudomonas aeruginosa-induced pneumonia model,we reported that IL-10-producing B cells(IL-10+B cells)play a key role in spontaneously resolving infection-mediated inflammation.Accumulated cytosolic reactive oxygen species(ROS)during inflammation were shown to drive IL-10+B-cell generation by remodeling one-carbon metabolism.Depletion of the enzyme serine hydroxymethyltransferase 1(Shmt1)led to inadequate one-carbon metabolism and decreased IL-10+B-cell production.Furthermore,increased one-carbon flux elevated the levels of the methyl donor S-adenosylmethionine(SAM),altering histone H3 lysine 4 methylation(H3K4me)at the Il10 gene to promote chromatin accessibility and upregulate Il10 expression in B cells.Therefore,the one-carbon metabolism-associated compound ethacrynic acid(EA)was screened and found to potentially treat infectious pneumonia by boosting IL-10+B-cell generation.Overall,these findings reveal that ROS serve as modulators to resolve inflammation by reprogramming one-carbon metabolism pathways in B cells.

Mutant p53 in cancer therapy-the barrier or the path

Since wild-type p53 is central for maintaining genomic stability and preventing oncogenesis, its coding gene TP53 is highly mutated in ~50% of human cancers, and its activity is almost abrogated in the rest of cancers. Approximately 80% of p53 mutations are single point mutations with several hotspot mutations. Besides loss of function and dominant-negative effect on the wild-type p53 activity, the hotspot p53 mutants also acquire new oncogenic functions, so-called ‘gain-of-functions’(GOF). Because the GOF of mutant p53 is highly associated with late-stage malignance and drug resistance, these p53 mutants have become hot targets for developing novel cancer therapies. In this essay, we review some recent progresses in better understanding of the role of mutant p53 GOF in chemoresistance and the underlying mechanisms, and discuss the pros and cons of targeting mutant p53 for the development of anti-cancer therapies.

SAMHD1对外源DNA的限制作用

本研究发现宿主限制因子SAMHD1在体外培养细胞和小鼠体内都表现出抑制外源DNA表达的活性.其中, SAMHD1对外源DNA的抑制作用不依赖其d NTPase和RNase活性,而且SAMHD1可以特异地结合干扰素调节因子1(Irf1)基因的启动子区域,从而上调IRF1的表达水平而抑制外源DNA的表达.进一步的实验研究发现,SAMHD1/IRF1能够显著降低外源DNA上与转录活化相关的组蛋白修饰的水平,并且SAMHD1/IRF1抑制外源DNA的表达不依赖于干扰素途径.在乙型肝炎病毒(HBV)细胞感染模型和HBV重组ccc DNA(rccc DNA)小鼠模型中,我们也证实了SAMHD1/IRF1对HBV的抗原表达和复制存在显著的抑制作用.本研究发现了限制外源DNA的表达是SAMHD1的一个新功能.

Circulating cell-free DNA for cancer early detection

Effective screening modalities are currently available for only a small subset of cancers,and they generally have suboptimal performance with complicated procedures.Therefore,there is an urgent need to develop simple,accurate,and non-invasive methods for early detection of cancers.Genetic and epigenetic alterations in plasma circulating cell-free DNA(cfDNA)have shown the potential to revolutionize methods of early detection of cancers and facilitate subsequent diagnosis to improve survival of patients.The medical interest in cfDNA assays has been inspired by emerging single-and multi-early detection of cancers studies.This review summarizes current technological and clinical advances,in the hopes of providing insights into the development and applications of cfDNA assays in various cancers and clinical scenarios.The key phases of clinical development of biomarkers are highlighted,and the future developments of cfDNA-based liquid biopsies in early detection of cancers are outlined.It is hoped that this study can boost the potential integration of cfDNA-based early detection of cancers into the current clinical workflow.

Phase separation—a strategy to resist autophagic degradation under heat stress

Stress spans our whole lives. When exposed to stressful conditions, every life, from a worm to a human, adjusts its internal properties to adapt to new environments. This principle applies to adults, and is also essential during development in animals. Dr. Zhang and coworkers from Institute of Biophysics, Chinese Academy of Sciences made a remarkable discovery showing that heat-stress-triggered accumulation of PGL granules is an adaptive response to maintain embryonic viability (Figure 1). Their work was published in the September 6th issue of Cell, entitled"mTOR regulates phase separation of PGL granules to modulate their autophagic degradation"(Zhang et al., 2018).