Pleiotrophin and N-syndecan promote perineural invasion and tumor progression in an orthotopic mouse model of pancreatic cancer

AIM To detect the expression of pleiotrophin(PTN) and N-syndecan in pancreatic cancer and analyze their association with tumor progression and perineural invasion(PNI).METHODS An orthotopic mouse model of pancreatic cancer was created by injecting tumor cells subcapsularly in a root region of the pancreas beneath the spleen. Pancreatic cancer tissues were taken from 36 mice that survived for more than 90 d. PTN and N-syndecan proteins were detected by immunohistochemistry and analyzed for their correlation with pathological features, PNI, and prognosis.RESULTS The expression rates of PTN and N-syndecan proteinswere 66.7% and 61.1%, respectively, in cancer tissue. PTN and N-syndecan expression was associated with PNI(P = 0.019 and P = 0.032, respectively). High PTN expression was closely associated with large bloody ascites(P = 0.009), liver metastasis(P = 0.035), and decreased survival time(P = 0.022). N-syndecan expression was significantly associated with tumor size(P = 0.025), but not with survival time(P = 0.539). CONCLUSION High PTN and N-syndecan expression was closely associated with metastasis and poor prognosis, suggesting that they may promote tumor progression and PNI in the orthotopic mouse model of pancreatic cancer.

Molecular imaging and therapy targeting coppermetabolism in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is the fifth most common cancer worldwide.Significant efforts have been devoted to identify new biomarkers for molecular imaging and targeted therapy of HCC.Copper is a nutritional metal required for the function of numerous enzymatic molecules in the metabolic pathways of human cells.Emerging evidence suggests that copper plays a role in cell proliferation and angiogenesis.Increased accumulation of copper ions was detected in tissue samples of HCC and many other cancers in humans.Altered copper metabolism is a new biomarker for molecular cancer imaging with position emission tomography(PET)using radioactive copper as a tracer.It has been reported that extrahepatic mouse hepatoma or HCC xenografts can be localized with PET using copper-64 chloride as a tracer,suggesting that copper metabolism is a new biomarker for the detection of HCC metastasis in areas of low physiological copper uptake.In addition to copper modulation therapy with copper chelators,short-interference RNA specific for human copper transporter 1(h Ctr1)may be used to suppress growth of HCC by blocking increased copper uptake mediated by h Ctr1.Furthermore,altered copper metabolism is a promising target for radionuclide therapy of HCC using therapeutic copper radionuclides.Copper metabolism has potential as a new theranostic biomarker for molecular imaging as well as targeted therapy of HCC.

Lymphocyte-to-monocyte ratio can predict mortality in pancreatic adenocarcinoma

AIMTo determine if the lymphocyte-to-monocyte ratio (LMR) could be helpful in predicting survival in patients with pancreatic adenocarcinoma. METHODSWe retrospectively reviewed the medical records of all patients diagnosed with pancreatic adenocarcinoma in the VA North Texas Healthcare System from January 2005 to December 2010. The LMR was calculated from peripheral blood cell counts obtained at the time of diagnosis of pancreatic cancer by dividing the absolute lymphocyte count by the absolute monocyte count. A Univariable Cox regression analysis was performed using these data, and hazard ratios (HR) and 95%CI were calculated. The median LMR (2.05) was used to dichotomize patients into high-LMR and low-LMR groups and the log rank test was used to compare survival between the two groups. RESULTSWe identified 97 patients with pancreatic adenocarcinoma (all men, 66% white, 30% African-American). The mean age and weight at diagnosis were 66.0 ± 0.9 (SEM) years and 80.4 ± 1.7 kg respectively. Mean absolute lymphocyte and monocyte values were 1.50 ± 0.07 K/μL and 0.74 ± 0.03 K/μL respectively. Mean, median and range of LMR was 2.36, 2.05 and 0.4-12 respectively. In the univariable Cox regression analysis, we found that an increased LMR was a significant indicator of improved overall survival in patients with pancreatic adenocarcinoma (HR = 0.83; 95%CI: 0.70-0.98; P = 0.027). Kaplan-Meier analysis revealed an overall median survival of 128 d (95%CI: 80-162 d). The median survival of patients in the high-LMR (> 2.05) group was significantly greater than the low-LMR group (≤ 2.05) (194 d vs 93 d; P = 0.03), validating a significant survival advantage in patients with a high LMR. CONCLUSIONThe LMR at diagnosis is a significant predictor for survival and can provide useful prognostic information in the management of patients with pancreatic adenocarcinoma.

Identification of host proteins that interact with African swine fever virus pE301R

African swine fever virus(ASFV)infection poses enormous threats and challenges to the global pig industry;however,no effective vaccine is available against ASFV,attributing to the huge viral genome(approximately189 kb)and numerous encoding products(>150 genes)due to the limited understanding on the molecular mechanisms of viral pathogenesis.Elucidating the host-factor/viral-protein interaction network will reveal new targets for developing novel antiviral therapies.Using proteomic analysis,we identified 255 cellular proteins that interact with the ASFV-encoded pE301R protein when transiently expressed in HEK293T cells.Gene ontology(GO)annotation,Kyoto Encyclopedia of Genes and Genomes(KEGG)database enrichment,and protein-protein interaction(PPI)network analyses revealed that pE301R-interacting host proteins are potentially involved in various biological processes,including protein translation and folding,response to stimulation,and mitochondrial transmembrane transport.The interactions of two putative cellular proteins(apoptosis inducing factor mitochondria associated 1(AIFM1)and vimentin(VIM))with pE301R-apoptosis inducing factor have been verified by co-immunoprecipitation.Our study revealed the inhibitory role of pE301R in interferon(IFN)induction that involves VIM sequestration by pE301R,identified interactions between ASFV pE301R and cellular proteins,and predicted the potential function of pE301R and its associated biological processes,providing valuable information to enhance our understanding of viral protein function,pathogenesis,and potential candidates for the prevention and control of ASFV infection.

Recent advances in the targeted fluorescent probes for the detection of metastatic bone cancer

Tumors of the breast,prostate,and lung are most likely to metastasize to the bone and typically indicates a poor cure and survival rate in cancer patients.Detection of metastatic bone cancer in early stage would save many lives and greatly improve patients’quality of life.Clinically,bone scintigraphy is often utilized to visualize bone metastases due to its relatively low cost and high sensitivity.Recently,a growth number of analytical researches aimed at developing targeted fluorescent probes to noninvasively image bone metastases with improved spatial resolution and specificity has been reported.In this review,we will summarize and discuss the recent published fluorescent probes on the accurate detection of metastatic bone cancer.First,the design principles of various targeted probes for imaging bone metastases will be presented,highlighting the signal moieties,targeting ligands,and physicochemical properties of the bone-specific probes.Next,the up-to-date bone-targeting fluorescent probes will be summarized and overviewed.Finally,future perspectives and challenges confronting the researchers in this field will be discussed.We believe this review will encourage novel ideas to develop smart targeted molecular probes for bone metastasis imaging,image-guided surgery,and therapeutic imaging materials.

Cancer and neurodegenerative disorders:pathogenic convergence through microRNA regulation

Although cancer and neurodegenerative disease are two distinct pathological disorders,emerging evidence indicates that these two types of disease share common mechanisms of genetic and molecular abnormalities.Recent studies show that individual microRNAs(miRNAs)could be involved in the pathology of both diseases,indicating that the mechanisms of these two seemingly dichotomous diseases converge in the dysregulation of gene expression at the post-transcriptional level.Given the increasing evidence showing that miRNA-based therapeutic strategies that modulate the activity of one or more miRNAs are potentially effective for a wide range of pathological conditions,the involvement of miRNAs in the common pathways of leading both diseases suggests a bright future for developing common therapeutic approaches for both diseases.Moreover,the miRNAs that are dysregulated in both diseases may hold promise as uniquely informative diagnostic markers.Here,we review recent studies on the miRNAs that have been implicated in both cancer and neurodegenerative diseases.

Therapeutic antibodies for precise cancer immunotherapy:current and future perspectives

Antibodies,as one of the most important components of host adaptive immune system,play an important role in defense of infectious disease,immune surveillance,and autoimmune disease.Due to the development of recombinant antibody technology,antibody therapeutics become the largest and rapidly expanding drug to provide major health benefits to patients,especially for the treatment of cancer patients.Many antibody-based therapeutic strategies have been developed including monoclonal anti-bodies,antibody-drug conjugates,bispecific and trispecific antibodies and pro-antibodies with promising results from both clinical and pre-clinical trials.However,the response rate and side-effect still vary between patients with undefined mechanisms.Here,we summarized the current and future perspectives of antibody-based cancer immunotherapeutic strategies for designing next-generation drugs.

MYC promotes global transcription in part by controlling P-TEFb complex formation via DNA-binding independent inhibition of CDK9 SUMOylation

MYC is an oncogenic transcription factor with a novel role in enhancing global transcription when overexpressed. However, how MYC promotes global transcription remains controversial. Here, we used a series of MYC mutants to dissect the molecular basis for MYC-driven global transcription. We found that MYC mutants deficient in DNA binding or known transcriptional activation activities can still promote global transcription and enhance serine 2 phosphorylation(Ser2P) of the RNA polymerase(Pol) II Cterminal domain(CTD), a hallmark of active elongating RNA Pol II. Two distinct regions within MYC can promote global transcription and Ser2P of Pol II CTD. The ability of various MYC mutants to promote global transcription and Ser2P correlates with their ability to suppress CDK9 SUMOylation and enhance positive transcription elongation factor b(P-TEFb) complex formation. We showed that MYC suppresses CDK9 SUMOylation by inhibiting the interaction between CDK9 and SUMO enzymes including UBC9 and PIAS1. Furthermore, MYC's activity in enhancing global transcription positively contributes to its activity in promoting cell proliferation and transformation. Together, our study demonstrates that MYC promotes global transcription, at least in part, by promoting the formation of the active P-TEFb complex via a sequence-specific DNA-binding activity-independent manner.

Microfluidic one-step, aqueous synthesis of size-tunable zeolitic imidazolate framework-8 for protein delivery

Zeolitic imidazolate framework-8(ZIF-8)with porous structure,biocompatibility,and pH-sensitive release behavior is a promising nanoplatform for protein delivery.However,it is still a challenging task for a practical synthesis of protein-loaded ZIF-8 nanoparticles.Here we report an all-aqueous microfluidic reactor for one-step,rapid,and highly controlled synthesis of ZIF-8 nanoparticles with high protein loading at room temperature.Microfluidic reactor allows for an ultrafast(<35 ms),complete mixing of Zn2+ions and 2-methylimidazole(2-MIM)at different molecular ratios,leading to the formation of stable ZIF-8 nanoparticles with tunable sizes(13.2–191.4 nm)in less than 30 s.By pre-mixing various proteins such as bovine serum albumin(BSA)(isoelectric point(pI)=5.82),ovalbumin(OVA)(pI=4.82),or RNase A(pI=8.93)with 2-MIM,ZIF-8 nanoparticles can be synthesized with protein encapsulation efficiency over 97%.Among the nanoparticles with different sizes,25 nm ZIF-8 nanoparticles show the best performance in promoting the cellular uptake of protein payload.Using OVA as a model protein,we demonstrate that 25 nm ZIF-8 nanoparticles significantly enhance the cytosolic delivery of antigen,as indicated by the effective activation of dendritic cells.We anticipate that this microfluidic synthesis of nanomaterials may advance the emerging field of cytosolic protein delivery.

Targeting tumor microenvironment with antibody-guided IL-2 pro-cytokine promotes and rejuvenates dysfunctional CD8^(+) T cells

Dear Editor,CLDN18.2(CLDN),a member of tight junction protein family,is strictly limited to express on differentiated epithelial cells of the gastric mucosa and abnormal overexpression has been found in many cancers,especially in digestive system malignancies.1 Those features make CLDN a potential therapeutic target.However,monoclonal antibody targeting CLDN induce limited antitumor immune responses in clinical trials and fusion of strong immunomodulators might be needed to enhance its efficacy.High dose IL-2 activates tumor infiltrating lymphocytes(TILs),but the severe toxicity and poor tumor targeting limits its use.

NAD（P）H：醌氧化还原酶1过表达在甲状腺髓样癌预后评估中的意义

甲状腺髓样癌（MTC）约占全部甲状腺癌的8％～10％，其临床表现复杂多样，易于局部扩散和远处转移，恶性程度高，预后差。因此，寻找MTC的早期诊断及预后评估的有效分子靶标是临床治疗MTC亟待解决的关键问题。NAD（P）H：醌氧化还原酶1（NQ01）可专一性的催化真核细胞内醌类物质及其衍生物发生双电子还原反应，从而对醌类物质的损伤形成一种保护机制。

醌氧化还原酶过表达在头颈部鳞状细胞癌预后评估中的意义

目的 探讨醌氧化还原酶（NQO1）过表达在头颈部鳞状细胞癌（HNSCC）中的临床预后评估意义.方法 采用免疫组织化学EnVision法检测NQO1蛋白在162例HNSCC、45例癌旁非瘤上皮和26例正常头颈部上皮组织中的表达,并分析其过表达与HNSCC患者预后之间的关系.结果 HNSCC组织中NQO1蛋白阳性率和强阳性率分别高达84.0％（136/162）和69.8％ （113/162）,均明显高于癌旁非瘤上皮和正常头颈部上皮组织（均P＜0.01）;卡方检验结果显示,NQO1蛋白表达水平与HNSCC的临床分期、分化程度和pT分期均密切相关（均P＜0.01）.Kaplan-Meier生存分析显示,NQO1蛋白高表达的HNSCC患者总生存期和无瘤生存期均明显低于NQO1蛋白低表达的患者（Log-rank分别为6.625和6.234,P分别为0.010和0.013）.Cox风险回归模型分析结果显示,NQO1蛋白高表达是影响HNSCC患者预后的独立风险因素（Wald=6.626,P=0.008）.结论 NQO1表达水平与HNSCC发生发展及预后密切相关.NQO1蛋白高表达可成为HNSCC患者不良预后的一个重要指标.

Targeting orthotopic gliomas with renal-clearable luminescent gold nanoparticles

A major clinical translational challenge in nanomedicine is the potential of toxicity associated with the uptake and long-term retention of non-degradable nanoparticles （NPs） in major organs. The development of inorganic NPs that undergo renal clearance could potentially resolve this significant biosafety concern. However, it remains unclear whether inorganic NPs that can be excreted by the kidneys remain capable of targeting tumors with poor permeability. Glioblastoma multiforme, the most malignant orthotopic brain tumor, presents a unique challenge for NP delivery because of the blood-brain barrier and robust blood-tumor barrier of reactive microglia and macroglia in the tumor microenvironment. Herein, we used an orthotopic murine glioma model to investigate the passive targeting of glutathione-coated gold nanoparticles （AuNPs） of 3 nm in diameter that undergo renal clearance and 18-nm AuNPs that fail to undergo renal clearance. Remarkably, we report that 3-nm AuNPs were able to target intracranial tumor tissues with higher efficiency （2.3× relative to surrounding non-tumor normal brain tissues） and greater specificity （3.0×） than did the larger AuNPs. Pharmacokinetics studies suggested that the higher glioma targeting ability of the 3-nm AuNPs may be attributed to the longer retention time in circulation. The total accumulation of the 3-nm AuNPs in major organs was significantly less （8.4×） than that of the 18-nm AuNPs. Microscopic imaging of blood vessels and renal-clearable AuNPs showed extravasation of NPs from the leaky blood-tumor barrier into the tumor interstitium. Taken together, our results suggest that the 3-nm AuNPs, characterized by enhanced permeability and retention, are able to target brain tumors and undergo renal clearance.

A Bayesian hierarchical model for analyzing methylated RNA immunoprecipitation sequencing data

Background： The recently emerged technology of methylated RNA immunoprecipitation sequencing （MeRIP-seq） sheds light on the study of RNA epigenetics. This new bioinformatics question calls for effective and robust peaking calling algorithms to detect mRNA methylation sites from MeRIP-seq data. Methods： We propose a Bayesian hierarchical model to detect methylation sites from MeRIP-seq data. Our modeling approach includes several important characteristics. First, it models the zero-inflated and over-dispersed counts by deploying a zero-inflated negative binomial model. Second, it incorporates a hidden Markov model （HMM） to account for the spatial dependency of neighboring read enrichment. Third, our Bayesian inference allows the proposed model to borrow strength in parameter estimation, which greatly improves the model stability when dealing with MeRIP-seq data with a small number of replicates. We use Markov chain Monte Carlo （MCMC） algorithms to simultaneously infer the model parameters in a de novo fashion. The R Shiny demo is available at https：//qiwei. shinyapps.io/BaySeqPeak and the R/C ＋＋ code is available at https：//github.com/liqiwei2000/BaySeqPeak. Results： In simulation studies, the proposed method outperformed the competing methods exomePeak and MeTPeak, especially when an excess of zeros were present in the data. In real MeRIP-seq data analysis, the proposed method identified methylation sites that were more consistent with biological knowledge, and had better spatial resolution compared to the other methods. Conclusions： In this study, we develop a Bayesian hierarchical model to identify methylation peaks in MeRIP-seq data. The proposed method has a competitive edge over existing methods in terms of accuracy, robustness and spatial resolution.

Design of synthetic materials for intracellular delivery of RNAs： From siRNA-mediated gene silencing to CRISPR/Cas gene editing

Ribonucleic acids （RNAs） possess great therapeutic potential and can be used to treat a variety of diseases. The unique biophysical properties of RNAs, such as high molecular weight, negative charge, hydrophilicity, low stability, and potential immunogenicity, require chemical modification and development of carriers to enable intracellular delivery of RNAs for clinical use. A variety of nanornaterials have been developed for the effective in vivo delivery of short/ small RNAs, messenger RNAs, and RNAs required for gene editing technologies including clustered regularly interspaced palindromic repeat （CRISPR）/Cas. This review outlines the challenges of delivering RNA therapeutics, explores the chemical synthesis of RNA modifications and carriers, and describes the efforts to design nanomaterials that can be used for a variety of clinical indications.