Identification of Prognosis-Related Genes and Key Target Genes for Pancreatic Cancer: A Bioinformatics Analysis

Objective: The mortality and morbidity rates associated with pancreatic cancer (PaCa) are extremely high. Various studies have demonstrated that pancreatic cancer will be the fourth cancer-related death by 2030, raising more concern for scholars to find effective methods to prevent and treat in order to improve the pancreatic cancer outcome. Using bioinformatic analysis, this study aims to pinpoint key genes that could impact PaCa patients’ prognosis and could be used as therapeutic targets. Methods: The TCGA and GEO datasets were integratively analyzed to identify prognosis-related differentially expressed genes. Next, the STRING database was used to develop PPI networks, and the MCODE and CytoNCA Cytoscape in Cytoscape were used to screen for critical genes. Through CytoNCA, three kinds of topology analysis were considered (degree, betweenness, and eigenvector). Essential genes were confirmed as potential target treatment through Go function and pathways enrichment analysis, a developed predictive risk model based on multivariate analysis, and the establishment of nomograms using the clinical information. Results: Overall, the GSE183795 and TCGA datasets associated 1311 and 2244 genes with pancreatic cancer prognosis, respectively. We identified 132 genes that were present in both datasets. The PPI network analysis using, the centrality analysis approach with the CytoNCA plug-in, showed that CDK2, PLK1, CCNB1, and TOP2A ranked in the top 5% across all three metrics. The independent analysis of a risk model revealed that the four key genes had a Hazard Ratio (HR) > 1. The monogram showed the predictive risk model and individual patient survival predictions were accurate. The results indicate that the effect of the selected vital genes was significant and that they could be used as biomarkers to predict a patient’s outcome and as possible target therapy in patients with pancreatic cancer. GO function and pathway analysis demonstrated that crucial genes might affect the P53 signaling pathway and FoxO signaling pathway, through which Meiotic nuclear division and cell cycle may have a significant function in essential genes affecting the outcome of patients who have pancreatic cancer. Conclusions: This study suggests that CDK2, CCNB1, PLK1 and TOP2A are four key genes that have a significant influence on PaCa migration and proliferation. CDK2, CCNB1, PLK1, and TOP2A can be used as potential PaCa prognostic biomarkers and therapeutic targets. However, experimental validation is necessary to confirm these predictions. Our study comes into contributions to the development of personalized target therapy for pancreatic cancer patients.

Phylogenetic analysis and target gene prediction of miR477 gene family in grape

To understand the molecular characteristics of the miR477 gene family of grape(Vvi-miR477)and to predict its target genes,the Vvi-miR477 genes were identified from previous small RNA sequencing data,then phylogenetic analysis and prediction of target gene were conducted.The Vvi-miR477 family consists of two precursor sequences and three mature sequences.The miR477 family members were mostly 19-22nt in length.The sequence is relatively conservative.Vvi-MIR477a and Vvi-MIR477b are located on chromosomes 1 and 2,respectively.These precursor sequences can form the typical stable stem-loop structure.Their minimum folding free energy is−39.10 kcal/mol and−50.90 kcal/mol,respectively.The MIR477 family can be divided into three groups.The prediction of target genes showed that Vvi-miR477 targets 26S proteasome,DEAD-box,GRAS family protein,Protein Phosphatase 2C,etc.The GO function of target genes was mainly enriched to six categories.The catabolic process,carboxylic ester hydrolase activity is shown to be high.This study provided a theoretical basis for further exploration of the molecular mechanism of miR477 in grape berry ripening.

Screening ulcerative colitis key target gene and pathway based on KEGG pathway

Objective:To obtain the relevant information of pathway and gene by using the database of DAVID,analyze the function and distribution of important genes,screen out the target genes related to Ulcerative Colitis and promote the study of the pathogenesis of UC and the development of new drugs.Methods:The Ulcerative Colitis was used to search UC related genes in TTD,Drugbank,DisGeNET database.The obtained gene data was input to the database of Daved,and the data of gene enriched pathway was obtained 87 genes were Significant enriched in the first 20 KEGG pathways.The 87 genes were input to the string database to make the interaction network diagram,and the key genes enriched in the pathway were also made the network diagram,and the two network diagrams were compared.Results:RELA,TNF,IL1B,NFKB1,IL6 and IL10 were among the highest ranked genes in two network diagrams.Conclusion:The study of RELA,TNF,IL1B,NFKB1,IL6 and IL10 is necessary for us to study further.The pathogenesis of UC was associated with multiple pathways such as NF-kappa B signaling pathway,Jak-STAT signaling pathway,TNF signaling pathway and so on.It is helpful to understand the pathogenesis of disease and provide a reliable target for the development of new drugs by analyzing the pathway and the network diagram of the interaction between genes and disease related genes.

Screening key target genes for pulmonary arterial hypertension based on bioinformatics

Background:Screening key target genes for pulmonary arterial hypertension(PAH)based on bioinformatics to provide a reference for the clinical development of drugs to cure PAH.Methods:The keyword“pulmonary arterial hypertension”was used to search related genes in the National Center for Biotechnology Information database(NCBI).The obtained genes data was input to the database of Database for Annotation,Visualization and Integrated Discovery(DAVID)(Version 6.8)to collect relevant information about pathways and genes.And the data of genes were enriched in 37 pathways and genes with occurrence frequency≥10 were respectively imported into the String database to construct protein-protein interaction(PPI)network diagrams,and the two network diagrams were compared.Results:VEGFA,MAPK1,MAPK3,IL6,JUN and TNF were among the highest-ranked genes in two network diagrams.Conclusion:The pathogenesis of PAH is associated with multiple pathways such as the TGF-βsignaling pathway,PI3K-Akt signaling pathway,MAPK signaling pathway,HIF-1 signaling pathway and so on.The study of VEGFA,MAPK1,MAPK3,IL6,JUN and TNF are closely related to PAH is necessary for us to study further.Through gene interaction network and pathway analysis of disease-associated genes,which will help us to screen the critical target genes of PAH and provide a reference for clinical development of effective drugs for PAH.

Microarray-based Screening of Target Genes Regulated by Heat Shock Factor AtHsfA1a in Arabidopsis thaliana

[ Objective] This study aimed to screen target genes regulated by heat shock factor AtHsfAla in Arabidopsis thaliana. [ Method] Using AtHsfAla-in- serted mutant athsfala （SALK-068042） and wild-type A. thaliana seedlings as experimental materials, target genes regulated by heat shock factor AtHsfAla were screened by microarray assay. Differentially expressed genes were screened by multiple method. Specific functions of differentially expressed genes were analyzed by gene ontology （GO） analysis. Signal transduction pathways, in which differentia｜ly expressed genes were involved, were analyzed by pathway analysis. Gene-gene interaction network was constructed by Signal-Net. [ Result] A total of 3 672 differentially expressed genes were screened out. Up-regulated differentially expressed genes were involved in 198 functions and 7 signal transduction pathways; down-regulated differentially expressed genes were involved in 94 functions and 10 signal transduction pathways. In the signal transduction network, it was found that cwlNV4 and HXK3 had relatively high ability of mediation; AT1 G14240 and cwlNV4 ex- hibited the most interactions with other genes, which were located in key positions throughout the gene-gene interaction network. [ Conclusion] Heat shock factor AtHsfAla regulates a large number of target genes in A. thaliana.

Identification and analysis of core target genes of miR-29b-3p in glioma

Objective:To investigate the core target genes of miR-29b-3p,and analyze the clinical significance of the core target genes in glioma.Methods:Bioinformatics analysis was used to predict and screen the target genes of miR-29b-3p.STRING and Cytoscape software were used to analyze the protein-protein interaction(PPI)of target genes.the differences expression and survival prognosis in glioma were analyzed by GEPIA and CGGA.Independent prognostic factors analyzed by univariate and multivariate Cox proportional hazards regression model.Results:22 target genes of miR-29b-3p were predicted using LinkedOmics,miRDB,miRTarBase,TargetScan,and starbase databases.Through the construction of the PPI network,genes out of the network were removed,and a total of 16 genes were screened for further study of their clinical significance.Based on analysis of GEPIA and CGGA databases,COL2A1,DNMT3A,and DNMT3B were excluded.Through further analysis of the univariate and multivariate Cox proportional hazard regression model,finally identified three core target genes:SERPINH1,LOXL2,CDK6.Conclusion:Bioinformatics analysis showed that miR-29b-3p targeted three core genes such as SERPINH1,LOXL2,and CDK6 in glioma.The expression of these genes was different between brain normal tissues and gliomas,between different grades of tumor,IDH mutation status and 1p/19q codeletion status.Its high expression had adverse effects on overall survival and recurrence-free survival.These core target genes can be used as an independent prognostic factor.

Screening key target genes for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)based on bioinformatics and gene network

Background:To provide a reference for the clinical development of drugs to suppress severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods:Retrieving genes related to SARS-CoV-2 with Genecards database and then importing the obtained gene data into the database of Database for Annotation,Visualization and Integrated Discovery(DAVID)(Version 6.8)to collect relevant information on pathways and genes.Genes enriched in the first 20 most significant pathways and genes with gene occurrence frequency≥6 were respectively imported into the STRING database to construct protein-protein interaction(PPI)network diagrams,and the two network diagrams were compared.Results:In the two network graphs,RELA,MAPK1,MAPK3,PIK3CA,PIK3R1,MAPK8,JAK1,STAT1,TNF,IL6,MAPK14,and IL1B ranked higher,and the occurrence frequency of the first 20 pathways was≥10.Conclusion:The pathogenesis of SARS-CoV-2 is associated with multiple pathways such as influenza A,TNF signaling pathway,chemokine signaling pathway,toll-like receptor signaling pathway,T cell receptor signaling pathway et al.RELA,MAPK1,MAPK3,PIK3CA,PIK3R1,MAPK8,JAK1,STAT1,TNF,IL6,MAPK14 and IL1B are closely related to SARS-CoV-2 and need further study.Gene interaction network and pathway analysis of diseaseassociated genes will help us to screen the key target genes of SARS-CoV-2 and provide a reference for the clinical development of effective drugs.

Polymorphisms of micro RNA target genes IL12B, INSR, CCND1 and IL10 in gastric cancer

AIM To evaluate associations between mi RNA target genes IL12B,INSR,CCND1 and IL10 polymorphisms and gastric cancer(GC)in European population.METHODS Gene polymorphisms were analyzed in 508 controls and474 GC patients from 3 tertiary centers in Germany,Lithuania and Latvia.Controls were patients from the out-patient departments,who were referred for upper endoscopy because of dyspeptic symptoms and had no history of previous malignancy.Gastric cancer(GC)patients had histopathological verification of gastric adenocarcinoma.Genomic DNA was extracted using salting out method from peripheral blood mononuclear cells.IL12B T>G(rs1368439),INSR T>C(rs1051690),CCND1 A>C(rs7177)and IL10 T>C(rs3024498)SNPs were genotyped by the real-time polymerase chain reaction.Associations between gene polymorphism and GC were evaluated using multiple logistic regression analysis with adjustment for sex,age and country of birth.RESULTS We observed similar distribution of genotypes and allelic frequencies of all polymorphisms between GC patients and controls except of INSR rs1051690.The frequency of the T allele of INSR gene was significantly higher in GC patients than in controls(23.26%and 19.19%respectively,P=0.028).CT genotype was also more prevalent in patients compared to control group(38.48%and 30.12%respectively,P<0.021).Logistic regression analysis revealed that only one polymorphism(rs1051690 in INSR gene)was associated with increased risk of GC.Carriers of CT genotype had higher odds of GC when compared to CC genotype(OR=1.45,95%PI:1.08-1.95,P=0.01).Similar association was observed in a dominant model for INSR gene,where comparison of TT+CT vs CC genotypes showed an increased risk of GC(OR=1.44,95%PI:1.08-1.90,P=0.01).Other analyzed SNPs were not associated with the presence of GC.CONCLUSION INSR rs1051690 SNP is associated with increased risk of GC,while polymorphisms in IL12B,CCND1 and IL10genes are not linked with the presence of GC.

Characterization of blueberry exosome-like nanoparticles and miRNAs with potential cross-kingdom human gene targets

Edible plant derived exosome-like nanoparticles(ELNs)have been shown to have multiple nutraceutical functions.However,the diversity of plant materials makes the plant derived ELN study challenging.More efforts are still needed to explore the feasible isolation methods of edible plant derived ELNs and the possible roles of food-derived ELNs in improving human health.In this study,a size exclusion chromatography based method was compared with the traditional ultracentrifugation method to isolate blueberry derived ELNs(B-ELNs),and the miRNA profile of B-ELNs was analyzed by high-throughput sequencing.A total of 36 miRNAs were found to be enriched in B-ELNs compared with berry tissue,and their potential cross-kingdom human gene targets were further predicted.Results showed that size exclusion chromatography was effective for B-ELN isolation.The most abundant miRNAs in B-ELNs mainly belonged to the miR166 family and miR396 family.Target gene prediction indicated that B-ELNs could potentially regulate pathways related to the human digestive system,immune system and infectious diseases.

To Analyze the Sensitivity of RT-PCR Assays Employing S Gene Target Failure with Whole Genome Sequencing Data during Third Wave by SARS-CoV-2 Omicron Variant

Introduction: Omicron is a highly divergent variant of concern (VOCs) of a severe acute respiratory syndrome SARS-CoV-2. It carries a high number of mutations in its spike protein hence;it is more transmissible in the community by immune evasion mechanisms. Due to mutation within S gene, most Omicron variants have reported S gene target failure (SGTF) with some commercially available PCR kits. Such diagnostic features can be used as markers to screen Omicron. However, Whole Genome Sequencing (WGS) is the only gold standard approach to confirm novel microorganisms at genetically level as similar mutations can also be found in other variants that are circulating at low frequencies worldwide. This Retrospective study is aimed to assess RT-PCR sensitivity in the detection of S gene target failure in comparison with whole genome sequencing to detect variants of Omicron. Methods: We have analysed retrospective data of SARS-CoV-2 positive RT-PCR samples for S gene target failure (SGTF) with TaqPath COVID-19 RT-PCR Combo Kit (ThermoFisher) and combined with sequencing technologies to study the emerged pattern of SARS-CoV-2 variants during third wave at the tertiary care centre, Surat. Results: From the first day of December 2021 till the end of February 2022, a total of 321,803 diagnostic RT-PCR tests for SARS-CoV-2 were performed, of which 20,566 positive cases were reported at our tertiary care centre with an average cumulative positivity of 6.39% over a period of three months. In the month of December 21 samples characterized by the SGTF (70/129) were suggestive of being infected by the Omicron variant and identified as Omicron (B.1.1.529 lineage) when sequence. In the month of January, we analysed a subset of samples (n = 618) with SGTF (24%) and without SGTF (76%) with Ct values Conclusions: During the COVID-19 pandemic, it took almost more than 15 days to diagnose infection and identify pathogen by sequencing technology. In contrast to that molecular assay provided quick identification with the help of SGTF phenomenon within 5 hours of duration. This strategy helps scientists and health policymakers for the quick isolation and identification of clusters. That ultimately results in a decreased transmission of pathogen among the community.

Identification of microRNA and analysis of target genes in Panax ginseng

Objective: Ginsenosides, polysaccharides and phenols, the main active ingredients in Panax ginseng, are not different significantly in content between 3 and 5 years old of ginsengs called Yuan ginseng and more than ten years old ones called Shizhu ginseng. The responsible chemical compounds cannot fully explain difference in efficacy between them. According to reports in Lonicerae Japonicae Flos(Jinyinhua in Chinese) and Glycyrrhizae Radix et Rhizoma(Gancao in Chinese), microRNA may play a role in efficacy,so we identified microRNAs in P. ginseng at the different growth years and analyzed their target genes.Methods: Using high-throughput sequencing, the RNA-Seq, small RNA-Seq and degradome databases of P. ginseng were constructed. The differentially expressed microRNAs was identified by qRT-PCR.Results: A total of 63,875 unigenes and 24,154,579 small RNA clean reads were obtained from the roots of P. ginseng. From these small RNAs, 71 miRNA families were identified by bioinformatics target prediction software, including 34 conserved miRNAs, 37 non-conserved miRNA families, as well as 179 target genes of 17 known miRNAs. Through degradome sequencing and computation, we finally verified 13 targets of eight miRNAs involved in transcription, energy metabolism, biological stress and disease resistance, suggesting the significance of miRNAs in the development of P. ginseng. Consistently, major miRNA targets exhibited tissue specificity and complexity in expression patterns.Conclusion: Differential expression microRNAs were found in different growth years of ginsengs(Shizhu ginseng and Yuan ginseng), and the regulatory roles and functional annotations of miRNA targets in P. ginseng need further investigation.

Gene targeted and immune therapies for nodal and gastrointestinal follicular lymphomas

Follicular lymphoma(FL)is the most common indolent B-cell lymphoma(BCL)globally.Recently,its incidence has increased in Europe,the United States,and Asia,with the number of gastrointestinal FL cases expected to increase.Genetic abnormalities related to t(14;18)translocation,BCL2 overexpression,NF-κB pathway-related factors,histone acetylases,and histone methyltransferases cause FL and enhance its proliferation.Meanwhile,microRNAs are commonly used in diagnosing FL and predicting patient prognosis.Many clinical trials on novel therapeutics targeting these genetic abnormalities and immunomodulatory mechanisms have been conducted,resulting in a marked improvement in therapeutic outcomes for FL.Although developing these innovative therapeutic agents targeting specific genetic mutations and immune pathways has provided hope for curative options,FL treatment has become more complex,requiring combinatorial therapeutic regimens.However,optimal treatment combinations have not yet been achieved,highlighting the importance of a complete understanding regarding the pathogenesis of gastrointestinal FL.Accordingly,this article reviews key research on the molecular pathogenesis of nodal FL and novel therapies targeting the causative genetic mutations.Moreover,the results of clinical trials are summarized,with a particular focus on treating nodal and gastrointestinal FLs.

Differential Gene Expression Profiles in Coronary Heart Disease Patients of Blood Stasis Syndrome in Traditional Chinese Medicine and Clinical Role of Target Gene

Objective:To investigate the differential gene expression profiles in coronary heart disease(CHD) patients of blood-stasis syndrome(BSS) by oligonucleotide microarray technique,and the clinical significance of target gene.Methods:Subjects were assigned to CHD patients with BSS(n=8),CHD patients without BSS (n=8),and BSS patients without CHD(n=8) based on coronary angiography and the diagnostic criteria of BSS. The sex- and age-matched healthy volunteers(n=8) were enrolled as the control group.Venous blood s...

A review of target gene specificity of flavonoid R2R3-MYB transcription factors and a discussion of factors contributing to the target gene selectivity

Flavonoid biosynthetic genes are often coordinately regulated in a temporal manner during flower or fruit development, resulting in specific accumulation profiles of flavonoid compounds. R2R3-MYB-type transcription factors （TFs） ＂recruit＂ a set of biosynthetic genes to produce flavonoids, and, therefore, R2R3-MYBs are responsible for the coordinated expression of structural genes. Although a wealth of information regarding the identified and functionally characterized R2R3-MYBs that are involved in flavonoid accumulation is available to date, this is the first review on the global regulation of MYB factors in the flavonoid pathway. The data presented in this review demonstrate that anthocyanin, flavone/flavonol/3-deoxyflavonoid （FFD）, proanthocyanidin （PA）, and isoflavonoid are independently regulated by different subgroups of R2R3-MYBs. Furthermore, FFD-specific R2R3-MYBs have a preference for early biosynthetic genes （EBGs） as their target genes; anthocyanin-specific R2R3-MYBs from dicot species essentially regulate late biosynthetic genes （LBGs）; the remaining R2R3-MYBs have a wider range of target gene specificity. To elucidate the nature of the differential target gene specificity between R2R3-MYBs, we analyzed the DNA binding domain （also termed the MYB-domain） of R2R3-MYBs and the distribution of the recognition cis-elements. We identified four conserved amino acid residues located in or just before helix-3 of dicot anthocyanin R2R3-MYBs that might account for the different recognition DNA sequence and subsequently the different target gene specificity to the remaining R2R3-MYB TFs.

The Pathogenesis and Treatment Progress of Androgenic Alopecia

Androgenic alopecia, also known as seborrheic alopecia, is the most common hair loss disorder in dermatology clinics, mainly characterized by hair follicle miniaturization and progressive hair loss. The etiology and pathogenesis of androgenic alopecia are not clear, but may be related to heredity and androgen metabolism. Currently, minoxidil and finasteride are the only two drugs approved by the U.S. Food and Drug Administration (FDA) for AGA treatment, other treatments include oral minoxidil, hair transplantation, low energy laser therapy (LLLT), platelet-rich plasma (PRP), Chinese medicine microneedles, and combination therapy. With the development of medicine and science, we have ushered in the era of biologics and targeted therapy. In recent years, a variety of signaling pathways for androgenic alopecia have been found, which may provide a basis for targeted therapy for androgenic alopecia.

Inhibitary effects of antisense oligonucleotide specific to K-ras point mutation on the target gene expression in human pancreatic carcinoma cells

The prognosis of pancreatic carcinoma is disappointing due to the difficulty of early and accurate diagnosis,low operative resection rate, insensitivity to radiation therapy and chemotherapy. The incidence of pancreatic carcinoma has increased considerably in the past few years, the progress of diagnosis and treatment, however, has not been significantly improved. The overall 5-year survive rate is still as low as 5%-10%, and the improved 5 years survive rate is about 20%-40% after successive Whipple-operation. The early diagnosis is critical to the successful surgical treatment. It depends on the establishment of the new way for that.

Characterization and target gene analysis of microRNAs in the antennae of the parasitoid wasp Microplitis mediator

MicroRNAs(miRNAs),a class of non-coding single-strand RNA molecules encoded by endogenous genes,are about 21–24 nucleotides long and are involved in the post-transcriptional regulation of gene expression in plants and animals.Generally,the types and quantities of miRNAs in the different tissues of an organism are diverse,and these divergences may be related to their specific functions.Here we have identified 296 known miRNAs and 46 novel miRNAs in the antennae of the parasitoid wasp Microplitis mediator by high-throughput sequencing.Thirty-three miRNAs were predicted to target olfactory-associated genes,including odorant binding proteins(OBPs),chemosensory proteins,odorant receptors(ORs),ionotropic receptors(IRs)and gustatory receptors.Among these,17 miRNAs were significantly highly expressed in the antennae,four miRNAs were highly expressed both in the antennae and head or wings,while the remaining 12 miRNAs were mainly expressed in the head,thorax,abdomen,legs and wings.Notably,miR-9a-5p and miR-2525-3p were highly expressed in male antennae,whereas miR-1000-5p and novel-miR-13 were enriched in female antennae.The 17 miRNAs highly expressed in antennae are likely to be associated with olfaction,and were predicted to target one OBP(targeted by miR-3751-3p),one IR(targeted by miR-7-5p)and 14 ORs(targeted by 15 miRNAs including miR-6-3p,miR-9a-5p,miR-9b-5p,miR-29-5p,miR-71-5p,miR-275-3p,miR-1000-5p,miR-1000-3p,miR-2525-3p,miR-6012-3p,miR-9719-3p,novel-miR-10,novel-miR-13,novel-miR-14 and novel-miR-28).These candidate olfactory-associated miRNAs are all likely to be involved in chemoreception through the regulation of chemosensory gene expression in the antennae of M.mediator.

Identification of Target Genes and Transcription Factors Implicated in Translation-Dependent Retrograde Sig- naling in Arabidopsis

Changes in organellar gene expression （OGE） trigger retrograde signaling. The molecular dissection of OGE-dependent retrograde signaling based on analyses of mutants with altered OGE is complicated by compensatory responses that mask the primary signaling defect and by secondary effects that influence other retrograde signaling pathways. Therefore, to identify the earliest effects of altered OGE on nuclear transcript accumulation, we have induced OGE defects in adult plants by ethanol-dependent repression of PRORS1, which encodes a prolyl-tRNA synthetase located in chloroplasts and mitochondria. After 32 h of PRORS1 repression, the translational capacity of chloroplasts was reduced, and this effect subsequently intensified, while basic photosynthetic parameters were still unchanged at 51 h. Analysis of changes in whole-genome transcriptomes during exposure to ethanol revealed that induced PRORS1 silencing affects the expression of 1020 genes in all. Some of these encode photosynthesis-related proteins, including several down-regulated light-harvesting chlorophyll a/b binding （LHC） proteins. Interestingly, genes for presumptive endoplasmic reticulum pro- teins are transiently up-regulated. Furthermore, several NAC-domain-containing proteins are among the transcription factors regulated. Candidate cis-acting elements which may coordinate the transcriptional co-regulation of genes sets include both G-box variants and sequence motifs with no similarity to known plant c/s-elements.

Genome-wide identification of target genes for transcription factor BR-C in the silkworm,Bombyx mori

Transcription factor Broad Complex(BR-C)is an ecdysone primary response gene in insects and participates in the regulation of insect growth and development.In this study,we performed a genome-wide identification of BR-C target genes in silkworm(Bombyx mori)using chromatin immunoprecipitation followed by high-throughput sequencing(ChIP-seq).As a result,a total of 1006 BR-C ChIP peaks were identified,and 15%of peaks were located in the promoter regions of 133 protein-coding genes.Functional annotation revealed that these ChIP peak-associated genes,as potential BR-C targets,were enriched in pathways related to biosynthetic process,metabolic process,and development.Transcriptome analysis and quantitative real-time polymerase chain reaction(PCR)examination revealed that developmental changes in expression patterns of a portion of potential BR-C targets,including HR96 and GC-otl,were similar to those of BR-C.ChIP-PCR examination confirmed that BR-C could directly bind to the promoters of potential targets.Further,dual luciferase assays demonstrated that HR96 promoter activity was significantly upregulated following BR-C overexpression,and this upregu-lation was abolished when the binding motif in the promoter was truncated.This study will be helpful for deciphering the regulatory roles of BR-C during insect growth and development.

Vitamin D receptor(VDR)contributes to the development of hypercalciuria by sensitizing VDR target genes to vitamin D in a genetic hypercalciuric stone-forming(GHS)rat model

Human idiopathic hypercalciuria(IH)is the most common cause of calcium oxalate nephrolithiasis with perturbed calcium metabolism with increased bone resorption and decreased renal calcium reabsorption,which can be phenotype-copied in the genetic hypercalciuric stone-forming(GHS)rat model.We previously demonstrated that high VDR expression plays important roles in the development of hypercalciuria in the GHS rats.However,the underlying mechanism through which VDR impact hypercalciuria development remains to be fully understood.Here,we sought to determine how VDR regulated its target genes that are implicated in calcium homeostasis and potentially hypercalciuria.We found that VDR expression in the GHS rats was elevated in the calcium transporting tissues,as well as in the thymus and prostate,but not in lung,brain,heart,liver and spleen,when compared with control SD rats.Snail expression in the GHS rats was significantly downregulated in kidney,intestine,thymus and testis.Intraperitoneal injection of 1,25(OH)2D3 significantly upregulated the expression of renal calcium sensing receptor(CaSR),intestinal calcium transporters transient receptor potential vanilloid type 6(TRPV6),and VDR in GHS rats,compared with that in control SD rats.ChIP assays revealed that VDR specifically bound to the proximal promoters of target genes,followed by histone H3 hyperacetylation or hypermethylation.Collectively,our results suggest that elevated VDR expressi on may con tribute to the development of hypercalciuria by sensi・tizing VDR target genes to 1,25(OH)2D3 through histone modifications at their promoter regions in a genetic hypercalciuric stone-forming(GHS)rat model.