Expression Of VCAN Research Articles

The oncogenic ADAMTS1–VCAN–EGFR cyclic axis drives anoikis resistance and invasion in renal cell carcinoma

BackgroundMetastasis, the leading cause of renal cell carcinoma (RCC) mortality, involves cancer cells resisting anoikis and invading. Until now, the role of the matrix metalloproteinase (MMP)-related enzyme, A disintegrin and metalloprotease with thrombospondin motifs 1 (ADAMTS1), in RCC anoikis regulation remains unclear.MethodsThe clinical significance of ADAMTS1 and its associated molecules in patients with RCC was investigated using data from the Gene Expression Omnibus (GEO) and TCGA datasets. Human phosphoreceptor tyrosine kinase (RTK) array, luciferase reporter assays, immunoprecipitation (IP) assays, western blotting, and real-time reverse-transcription quantitative polymerase chain reaction (RT–qPCR) were used to elucidate the underlying mechanisms of ADAMTS1. Functional assays, including anoikis resistance assays, invasion assays, and a Zebrafish xenotransplantation model, were conducted to assess the roles of ADAMTS1 in conferring resistance to anoikis in RCC.ResultsThis study found elevated ADAMTS1 transcripts in RCC tissues that were correlated with a poor prognosis. ADAMTS1 manipulation significantly affected cell anoikis through the mitochondrial pathway in RCC cells. Human receptor tyrosine kinase (RTK) array screening identified that epidermal growth factor receptor (EGFR) activation was responsible for ADAMTS1-induced anoikis resistance and invasion. Further investigations revealed that enzymatically active ADAMTS1-induced versican V1 (VCAN V1) proteolysis led to EGFR transactivation, which in turn, through positive feedback, regulated ADAMTS1. Additionally, ADAMTS1 can form a complex with p53 to influence EGFR signaling. In vivo, VCAN or EGFR knockdown reversed ADAMTS1-induced prometastatic characteristics of RCC. A clinical analysis revealed a positive correlation between ADAMTS1 and VCAN or the EGFR and patients with RCC with high ADAMTS1 and VCAN expression had the worst prognoses.ConclusionsOur results collectively uncover a novel cyclic axis involving ADAMTS1–VCAN–EGFR, which significantly contributes to RCC invasion and resistance to anoikis, thus presenting a promising therapeutic target for RCC metastasis.

Construction of a panoramic mRNA map of adult noncystic fibrosis bronchiectasis and a preliminary study of the underlying molecular mechanisms

BackgroundThe pathogenesis of noncystic fibrosis bronchiectasis in adults is complex, and the relevant molecular mechanisms remain unclear. In this study, we constructed a panoramic map of bronchiectasis mRNA, explored the potential molecular mechanisms, and identified potential therapeutic targets, thus providing a new clinical perspective for the preventive management of bronchiectasis and its acute exacerbation.MethodsThe mRNA profiles of peripheral blood and bronchiectasis tissues were obtained through transcriptome sequencing and public databases, and bioinformatics methods were used to screen for differentially expressed genes (DEGs). The DEGs were then subjected to biological function and pathway analyses. Some DEGs were validated using a real-time quantitative polymerase chain reaction (RT-qPCR) in peripheral blood. Spearman’s correlation analysis was used to analyse the correlation between DEGs and clinical indicators.ResultsBased on transcriptome sequencing and public databases, the mRNA profile of bronchiectasis was determined. DEGs were obtained from the peripheral blood sequencing dataset (985 DEGs), tissue sequencing dataset (2919 DEGs), and GSE97258 dataset (1083 DEGs). Bioinformatics analysis showed that upregulated DEGs had enriched neutrophil-related pathways, and downregulated DEGs had enriched ribosome-related pathways. RT-qPCR testing confirmed the upregulated expression of VCAN, SESTD1, SLC12A1, CD177, IFI44L, SIGLEC1, and RSAD2 in bronchiectasis. These genes were related to many clinical parameters, such as neutrophils, C-reactive protein, and procalcitonin (P < 0.05).ConclusionsTranscriptomic methods were used to construct a panoramic map of bronchiectasis mRNA expression. The findings showed that neutrophil activation, chronic inflammation, immune regulation, impaired ribosomal function, oxidative phosphorylation, and energy metabolism disorders are important factors in the development of bronchiectasis. VCAN, SESTD1, SLC12A1, CD177, IFI44L, SIGLEC1, and RSAD2 may play important roles in the pathogenesis of bronchiectasis and are potential therapeutic targets.

Single-cell transcriptome in the examination of the molecular characteristics of macrophages in lung cancer brain metastasis.

e20546 Background: Brain metastasis is one of the most common distant metastasis types in lung cancer patients. Its mechanism involves multiple factors, including cell-cell interactions, immune evasion, and the influence of the microenvironment. Single-cell RNA sequencing technology has significantly advanced our understanding of tumors at a single-cell resolution. It enables an in-depth understanding of the expression profiles of metastatic cells in brain tissue, providing potential predictive biomarkers for future monitoring, diagnosis, and effective treatment. Methods: We collected 50 samples from the GEO database, including 17 normal lung samples, 17 lung cancer samples, and 16 brain metastasis samples. We used the MNN method for batch correction and performed differential analysis using findAllMarkers and FindMarkers. Additionally, we conducted enrichment analysis using ClusterProfiler. Results: After batch correction and re-clustering of the macrophages, we identified 9 clusters. Based on their biological functions, they were classified into 5 macrophage subtypes: RTM_macro, Angio_macro, SEPP1_macro, Prolif_macro, and Mono_like. The angiogenic macrophages (Angio_macro) were most prominent in lung cancer brain metastasis samples (52%), followed by lung cancer (25%), and least present in normal samples (17%). Enrichment analysis revealed that iron death, antigen processing and presentation, and endoplasmic reticulum protein processing were more enriched in brain metastasis. This correlates with the high proportion of angiogenic macrophages present in the brain. The SEPP1_macro subtype showed high expression of SEPP1 and VCAN, genes associated with angiogenesis. Studies suggest that SEPP1 is associated with poor prognosis as it promotes neovascularization and tumor invasion. This subtype was highly expressed in brain metastasis (40%) and lung cancer (45%), but rare in normal samples (4%). In the SEPP1_macro subtype, brain metastasis samples were enriched in signaling pathways such as tumor necrosis factor, PI3K-AKT, NF-kB, and Mitogen-activated protein kinase (MAPK), focusing mainly on the interaction of cytokines and cytokine receptors. Conclusions: In lung cancer and lung cancer brain metastasis, there is a significant presence of macrophages that promote angiogenesis and subpopulations of tumor-promoting cells associated with poor prognosis. The existence of these two cell subsets may indicate their activation during tumor metastasis and their regulatory role in the formation of new blood vessels, thereby promoting tumor growth.

Biochemical profiling of the follicular environment to predict oocyte competence in cattle.

To identify markers of oocyte competence, we compared the biochemical characteristics of fluid and cells from follicles containing oocytes with different capacities to form an embryo. Follicles (5-6 mm) were dissected, and follicular fluid (FF), granulosa cells (GC), cumulus cells (CC) from immature and mature cumulus-oocyte-complexes (COC) were individually collected. The oocytes were matured, fertilized, and cultured individually until day 8 (D8) of development. On D8, the samples were grouped according to embryo production into those that gave rise to blastocysts (EMB) and those that did not reach the blastocyst stage (NEMB). In CCs from immature and mature COCs and GCs, expression of CASP3, SERPINE2, VCAN, LUM, FSHR, EGFR, PGR, and GHR genes was quantified. Cell-free DNA (cfDNA), progesterone, and estradiol concentrations in the FF were determined. Data were analyzed by Mann-Whitney U test (GraphPad Prism 9). GHR was highly expressed in immature CCs from the EMB group, whereas CASP3 was highly expressed in mature CCs from the NEMB group (P<0.05). During maturation, the expression of CASP3 and GHR genes increased only in the NEMB group. ART2 cfDNA was highly detected in FF of the NEMB compared to the EMB group. Progesterone concentration was similar between the groups, whereas estradiol concentration was higher (P<0.05) in the EMB than in the NEMB group. It was concluded that a higher level of GHR transcripts in immature CCs, lower CASP3 expression in CCs from matured COCs, lower levels of ART2, and higher estradiol concentrations in FF may indicate oocytes with greater potential for development.

CircBRAF promotes the progression of triple-negative breast cancer through modulating methylation by recruiting KDM4B to histone H3K9me3 and IGF2BP3 to mRNA.

Understanding the molecular characteristics of triple-negative breast cancer (TNBC) and developing more tailored treatment approaches is crucial. Circular RNAs (circRNAs), as potential therapeutic targets, remain largely unexplored in TNBC. This study utilized circRNA microarray analysis to determine the expression of circRNAs in TNBC, analyzing nine patient specimens. The characteristics of circBRAF were examined using divergent PCR primers, Sanger sequencing, fluorescence in situ hybridization (FISH) analysis, and the application of RNase and actinomycin D. The biological function of circBRAF in TNBC was further investigated through colony formation, tube formation, and transwell assays. Crucially, the mechanisms underlying the effects of circBRAF on TNBC progression were explored via RNA immunoprecipitation sequencing (RIP-seq) data, MS2 pulldown, RNA sequencing (RNA-seq) analysis, circBRAF knockdown, histone H3K9me3 modification, and Chromatin Isolation by RNA Purification (ChIRP) tests followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). We focused particularly on hsa_circ_0007178, produced from exons 4-13 of the oncogene BRAF. Functional experiments revealed that circBRAF is crucial for the development of TNBC, with its knockdown preventing angiogenesis, metastasis, and cell division in vitro. Mechanistically, circBRAF interacts with KDM4B and IGF2BP3, promoting TNBC growth. Interaction of circBRAF with IGF2BP3 increased the expression of VCAN, FN1, CDCA3, or B4GALT3 by controlling mRNA stability through RNA N6-methyladenosine (m6A) modification. Furthermore, circBRAF upregulated the expression of ADAMTS14 and MMP9 through recruitment of KDM4B to enhance respective H3K9me3 modification. Furthermore, overexpression of circBRAF was able to overcome the inhibitory effects of siKDM4B and siIGF2BP3 on cell migration and invasion. Our findings suggest that circBRAF may act as an oncogene in TNBC through its specific interactions with KDM4B and IGF2BP3, implying that circBRAF could serve as a potentially effective novel therapeutic target for TNBC.

Identification of tumor-agnostic biomarkers for predicting prostate cancer progression and biochemical recurrence.

The diverse clinical outcomes of prostate cancer have led to the development of gene signature assays predicting disease progression. Improved prostate cancer progression biomarkers are needed as current RNA biomarker tests have varying success for intermediate prostate cancer. Interest grows in universal gene signatures for invasive carcinoma progression. Early breast and prostate cancers share characteristics, including hormone dependence and BRCA1/2 mutations. Given the similarities in the pathobiology of breast and prostate cancer, we utilized the NanoString BC360 panel, comprising the validated PAM50 classifier and pathway-specific signatures associated with general tumor progression as well as breast cancer-specific classifiers. This retrospective cohort of primary prostate cancers (n=53) was stratified according to biochemical recurrence (BCR) status and the CAPRA-S to identify genes related to high-risk disease. Two public cohort (TCGA-PRAD and GSE54460) were used to validate the results. Expression profiling of our cohort uncovered associations between PIP and INHBA with BCR and high CAPRA-S score, as well as associations between VCAN, SFRP2, and THBS4 and BCR. Despite low levels of the ESR1 gene compared to AR, we found strong expression of the ER signaling signature, suggesting that BCR may be driven by ER-mediated pathways. Kaplan-Meier and univariate Cox proportional hazards regression analysis indicated the expression of ESR1, PGR, VCAN, and SFRP2 could predict the occurrence of relapse events. This is in keeping with the pathways represented by these genes which contribute to angiogenesis and the epithelial-mesenchymal transition. It is likely that VCAN works by activating the stroma and remodeling the tumor microenvironment. Additionally, SFRP2 overexpression has been associated with increased tumor size and reduced survival rates in breast cancer and among prostate cancer patients who experienced BCR. ESR1 influences disease progression by activating stroma, stimulating stem/progenitor prostate cancer, and inducing TGF-β. Estrogen signaling may therefore serve as a surrogate to AR signaling during progression and in hormone-refractory disease, particularly in prostate cancer patients with stromal-rich tumors. Collectively, the use of agnostic biomarkers developed for breast cancer stratification has facilitated a precise clinical classification of patients undergoing radical prostatectomy and highlighted the therapeutic potential of targeting estrogen signaling in prostate cancer.

DSE inhibits melanoma progression by regulating tumor immune cell infiltration and VCAN

Dermatan sulfate epimerase (DSE) is a C5 epiminase that plays a key role in converting chondroitin sulfate into dermal sulfate. DSE is often upregulated during carcinogenesis of some types of cancer and can regulate growth factor signaling in cancer cells. However, the expression and function of DSE in human melanoma have not been reported. In this study, we investigated the influence of tumor-derived DSE in melanoma progression and the potential mechanism of their action. First, proteomic analysis of collected melanoma tissues revealed that DSE was significantly down-regulated in melanoma tissues. DSE silenced or overexpressed melanoma cells were constructed to detect the effect of DSE on melanoma cells, and it was found that the up-regulation of DSE significantly inhibited the proliferation, migration and invasion of melanoma cells. Data analysis and flow cytometry were used to evaluate the immune subpopulations in tumors, and it was found that the high expression of DSE was closely related to the invasion of killer immune cells. Mechanistically, DSE promoted the expression of VCAN, which inhibited the biological activity of melanoma cells. Together, these results suggest that DSE is downregulated in melanoma tissues, and that high expression of DSE can promote melanoma progression by inducing immune cell infiltration and VCAN expression.

Identification of mutation gene prognostic biomarker in multiple myeloma through gene panel exome sequencing and transcriptome analysis in Chinese population

BackgroundThe 5-year survival rate of multiple myeloma (MM) in China is less than 40%, with considerable individual heterogeneity. Gene mutations are important predictive biomarkers that influence MM treatment decision. The aim of our study was to uncover the clinical significance of mutated genes in MM in the Chinese population. MethodsTargeted exon panel sequencing was performed of 400 genes to detect the gene mutation status in plasma cells from 50 patients with MM. DAVID was used to explore the functions and pathways of mutated genes. Detection of mutant gene expression, prognosis and immune cell infiltration with GSE6477. GEO2R was utilized to identify differentially expressed genes (DEGs). Kaplan-Meier and CIBERSORT were applied to compare survival distributions and evaluate the gene expression associated with immune cell infiltration, respectively. ResultsMutations of 337 genes were identified in MM. The mutation types included SNP, INS, and DEL, but the dominant mutation type was SNP. Function and pathway analysis of mutant genes were performed to elucidate DNA modifications. We identified a total number of 660 downregulated and 587 upregulated genes from the GSE6477 dataset. Thirty-three common genes were present in both the mutant genes and DEGs. The functions and pathways of the mutated genes were enriched in myeloid cell differentiation, regulation of hemopoiesis, etc. Moreover, we found that the low expression of BCL6, BIRC3, HLA-DQA1, and VCAN was correlated with poor prognosis in MM. ConclusionsThe mutations and low expression of BCL6, BIRC3, HLA-DQA1, and VCAN were correlated with poor prognosis and immune cell infiltration in MM. This study is the first to reveal the spectrum of mutations in the Chinese population by the use of an NGS panel.

O-097 Predicting live birth rate using combined Day3-spent embryo culture medium metabolomics and cumulus cell gene expression based algorithm to optimize embryo selection in unexplained infertility

Abstract Study question Can combination of metabolomics of spent embryo culture medium (sECM) and cumulus cell (CC) genes judge live birth rate in unexplained infertility (UI)? Summary answer A combination of day3 metabolic profiling of sECM with CC gene expression yields an algorithm to predict live birth rate in UI. What is known already Optimizing embryo selection is rate-limiting for pregnancy success. The metabolomic profile of sECM can aid in advanced prediction of embryonic developmental potential. Recent transcriptomics studies proposed prokinectin 2 (PROK2), and pregnancy up-regulated nonubiquitous CaM kinase (PNCK) as two novel gene/s responsible for perifollicular vascularization and embryonic development respectively; two important events regulating bidirectional CC-oocyte signalling and implantation ability. Given that UI is contributory to impaired oocyte quality, we delineated if combined metabolic signature of CC gene expression encompassing metabolic, developmental competence, and extracellular matrix facet/s and sECM could address live birth rate in women with UI following single embryo transfer. Study design, size, duration A prospective cohort study was conducted at Institute of Reproductive Medicine between January 2022 and October 2022. 48 women (age: 30-39 years) diagnosed with UI undergoing oocyte-retrieval offered consented participation and were sub-stratified according to pregnancy outcome. Cleavage-stage embryos from intra cytoplasmic sperm injection (ICSI) cycles were scored according to combination of gene expression pattern of CC-samples and integral value of metabolite obtained from nuclear magnetic resonance (NMR) spectroscopy of sECM (range 0-3, 4-6, 7-9). Participants/materials, setting, methods Expression profiles of metabolic (LDHA, PFKP, PKM2), extracellular matrix (HAS2, PTX3, TNFAIP6, VCAN), and developmental competence (PNCK, PROK2) related genes were evaluated by real time-quantitative polymerase-chain-reaction (qRT-PCR) from retrieved CC. Day-3 sECM (30 μL) was collected for NMR from 43 single transferred embryos. Univariate and multivariate analysis was performed after NMR data acquisition. Live birth rate was evaluated by area-under-the-curve (AUC) values using combined score panel. Main results and the role of chance The groups were similar for patient age, BMI, AMH, infertility diagnosis, and stimulation protocol. A total of 43 cycles were included in the analysis [embryo transfer cancelled (n = 2), insufficient cumulus mass (n = 3)]. Clinical pregnancy was confirmed in 16 patients (37.21%). Mean mRNA levels for all genes were similar in CC associated with oocyte that fertilized normally (2PN) compared with those that either failed to fertilize or were abnormal (i.e. 1PN, 3PN). Similar results were observed in terms of speed of embryonic cleavage (≥7 cell on Day 3 vs. ≤6 cell on Day 3). Expression of VCAN, (OR: 1.102, 95% CI: 1.010-1.202, p &amp;lt; 0.02), PNCK (OR: 0.931, 95% CI: 0.916-0.947, p &amp;lt; 0.001) and PROK2 (OR: 1.273, 95% CI: 1.011-1.231, p &amp;lt; 0.001) was significantly higher (p &amp;lt; 0.01) in CC that achieved live birth compared with those that failed to result a successful pregnancy. Multivariate and univariate analysis revealed distinct metabolomic signatures between pregnant and non- pregnant group/s. Lactate, pyruvate and proline were the most significant (p &amp;lt; 0.01) altered metabolite in sECM of non-pregnant group when compared to their pregnant counterpart. The combination demonstrated a “good” (VCAN, pyruvate: AUC: 0.87), “fair” (PROK2, proline: AUC: 0.79) and “modest” (PNCK, glucose: AUC: 0.71) in terms of live-birth rate. Limitations, reasons for caution The study population is heterogeneous in terms of duration of infertility and previous infertility treatment outcome, limited by small sample size and subjective nature of embryo scoring. The presumable involvement of unknown factors in developmental competence of UI and impact of frozen cycle/s should also be addressed. Wider implications of the findings The study provides reliable accuracy measures to support relationship between live birth rate and combined score panel in patients with UI. It is envisioned integration of CC gene expression/s and metabolomics of sECM if combined with patient characteristics may tailor therapy by artificial intelligence to model the outcome in UI. Trial registration number Not applicable

CXCL10 mediates CD8+ T cells to facilitate vessel normalization and improve the efficacy of cetuximab combined with PD-1 checkpoint inhibitors in colorectal cancer

The immunotherapy and anti-EGFR targeted treatment occupying a pivotal position in colorectal cancer (CRC), is still limited to a group of patients who display specific molecular alterations and inevitably escape from resistance, further studies are still needed in colorectal cancer. We found that chemokine ligand 10 (CXCL10) expression correlates with intratumoral CD8+ T cell infiltration and reprograms tumor vasculatures in colorectal cancer. CXCL10 overexpression not only suppressed tumor growth but also increased CD8+ T cell infiltration and induced tumor vascular normalization in vivo. Additionally, the growth inhibition and tumor vascular normalization induced by CXCL10 can be reversed by the depletion of CD8+ T cells in vivo. Mechanically, CXCL10 interacts with VCAN to mediate tumor vascular normalization. The VCAN expression correlated inversely with the expression of CXCL10 and the infiltration of CD8+ T cells in CRC. Elevated CXCL10 expression sensitized colorectal cancer cells to cetuximab/anti-PD1 combination therapy compared with cetuximab or anti-PD1 alone. We propose that CXCL10 could be used to increase the anti-EGFR therapy and immunotherapy effect, targeting both tumor vessels and immune cells in colorectal cancer.

A comprehensive assessment of VCAN transcriptional expression and evaluation as an effective prognostic biomarker against breast cancer: in silico study

BackgroundThe VCAN gene provides instructions for making a protein called versican which is a type of protein known as a proteoglycan. Versican is a key ingredient of the extracellular matrix, and due to its widespread expression in the body, versican is involved in cell adhesion, proliferation, and migration. Mutations or alterations of this protein could result in the disintegration of the fine-tuned molecular machinery which can lead to uncontrolled cell proliferation.ResultsVCAN is a novel prognostic marker for multiple cancers, and it showed tremendous results on breast cancer prognosis based on the data available on multiple websites. So, we targeted VCAN to analyze the expression and the outcome of breast cancer. This is a server-based study, and the expression of VCAN shows upregulation in breast cancer subtypes as compared to the normal tissue. The promoter methylation analysis suggested that overexpression of VCAN may be due to hypomethylation. Mutation analysis showed a positive correlation with VCAN expression where missense-type mutation has the highest percentage (77.33%), truncating (17.33%), and splice (4%) and somatic mutation frequency is 1.8%. VCAN was closely related to ten different genes and coexpressed with five of the genes among them. Five distinct compounds are linked to the methylation and mutagenesis of VCAN, according to the gene–drug interaction.ConclusionsThe upregulation of VCAN is closely correlated with promoter methylation and the clinical features of breast cancer patients. The whole study suggests that the breast cancer patient’s survival rate gets lower when the VCAN expression level gets higher. We anticipated that these findings will lead to further improvements in breast cancer prognosis and the significance of VCAN as a biomarker for breast cancer prognosis.

Joint effect of THBS2 and VCAN accelerating the poor prognosis of gastric cancer.

Gastric cancer is the most common malignant tumor of the digestive system. The progression from gastritis to gastric cancer may be related to genetic factors, but the specific molecular mechanism remains unclear. Therefore, an in-depth study of the molecular mechanism of gastritis and gastric cancer is significant. We downloaded two gene profiles, GSE2669 and GSE116312, from the Gene Expression Omnibus (GEO) database. This study aims to apply bioinformatics technology to mine differentially expressed genes (DEGs), DEGs annotation, protein-protein interaction (PPI) network creation, and hub gene identification and expression between gastric cancer patients and gastritis patients. Overall survival analysis of hub genes, analysis by comparative toxicogenomics database for hub genes in gastric cancer, THBS2 and VCAN protein expression by immunohistochemistry for gastric cancer and gastritis as well as design of the biological process (BP) neural network was implemented. The MSLN, SPP1, THBS2, SPARC, FN1, IGFBP7, VCAN were up-regulated in gastric carcinoma samples, while FGA was down-regulated. The protein expression of THBS2 and VCAN in gastric cancer was significantly higher than that in gastritis. VCAN protein expression was positively associated with tumor invasion (P = 0.011) and HER2 overexpression (P = 0.031). Strong correlation among THBS2, VCAN, and gastric cancer based on the BP neural network. THBS2 and VCAN may be potential targets for improving gastric cancer patients' diagnosis and clinical efficacy.

VCAN, expressed highly in hepatitis B virus-induced hepatocellular carcinoma, is a potential biomarker for immune checkpoint inhibitors.

BACKGROUNDAs a proteoglycan, VCAN exists in the tumor microenvironment and regulates tumor proliferation, invasion, and metastasis, but its role in hepatocellular carcinoma (HCC) has not yet been elucidated.AIMTo investigate the expression and potential mechanism of action of VCAN in HCC.METHODSBased on The Cancer Genome Atlas Liver Hepatocellular Carcinoma dataset, we explored the correlation between VCAN expression and clinical features, and analyzed the prognosis of patients with high and low VCAN expression. The potential mechanism of action of VCAN was explored by Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes analysis, and gene set enrichment analysis. We also explored immune cell infiltration, immune checkpoint gene expression, and sensitivity of immune checkpoint [programmed cell death protein 1 (PD-1)/cytotoxic T lymphocyte antigen 4 (CTLA4)] inhibitor therapy in patients with different VCAN expression. VCAN mRNA expression and VCAN methylation in peripheral blood were tested in 100 hepatitis B virus (HBV)-related patients (50 HCC and 50 liver cirrhosis).RESULTSVCAN was highly expressed in HCC tissues, which was associated with a poor prognosis in HCC patients. No significant difference was found in VCAN mRNA expression in blood between patients with HBV-related cirrhosis and those with HCC, but there was a significant difference in VCAN methylation between the two groups. The correlation between VCAN and infiltrations of several different tumor immune cell types (including B cells, CD8+ T cells, and eosinophils) was significantly different. VCAN was strongly related to immune checkpoint gene expression and tumor mutation burden, and could be a biomarker of sensitivity to immune checkpoint (PD1/CTLA4) inhibitors. In addition, VCAN mRNA expression was associated with hepatitis B e antigen, HBV DNA, white blood cells, platelets, cholesterol, and coagulation function.CONCLUSIONHigh VCAN level could be a possible biomarker for poor prognosis of HCC, and its immunomodulatory mechanism in HCC warrants investigation.

Landscape and age dynamics of immune cells in the Egyptian rousette bat.

Bats harbor high-impact zoonotic viruses often in the absence of disease manifestation. This restriction and disease tolerance possibly rely on specific immunological features. In-depth molecular characterization of cellular immunity and imprinting of age on leukocyte compartments remained unexplored in bats. We employ single-cell RNA sequencing (scRNA-seq) and establish immunostaining panels to characterize the immune cell landscape in juvenile, subadult, and adult Egyptian rousette bats (ERBs). Transcriptomic and flow cytometry data reveal conserved subsets and substantial enrichments of CD79a+ B cells and CD11b+ Tcells in juvenile animals, whereas neutrophils, CD206+ myeloid cells, and CD3+ Tcells dominate as bats reach adulthood. Despite differing frequencies, phagocytosis of circulating and tissue-resident myeloid cells and proliferation of peripheral and splenic lymphocytes are analogous in juvenile and adult ERBs. We provide a comprehensive map of the immune landscape in ERBs and show age-imprinted resilience progression and find that variability in cellular immunity only partly recapitulates mammalian archetypes.

Identification of Candidate Therapeutic Target Genes and Profiling of Tumor-Infiltrating Immune Cells in Pancreatic Cancer via Integrated Transcriptomic Analysis.

Pancreatic cancer (PC) has a dismal prognosis despite advancing scientific and technological knowledge. The exploration of novel genes is critical to improving current therapeutic measures. This research is aimed at selecting hub genes that can act as candidate therapeutic target genes and as prognostic biomarkers in PC. Gene expression profiles of datasets GSE101448, GSE15471, and GSE62452 were extracted from the GEO database. The “limma” package was performed to select differentially expressed genes (DEGs) between PC and normal tissue samples in each dataset. Robust rank aggregation (RRA) algorithm was conducted to integrate multiple expression profiles and identify robust DEGs. GO analysis and KEGG analysis were conducted to identify the functional correlation of the DEGs. The CIBERSORT algorithm was conducted to estimate the immune cell composition of each tissue sample. STRING and Cytoscape were used to establish the protein-protein interaction (PPI) network. The cytoHubba plugin in Cytoscape was performed to identify hub genes. Survival analysis based on hub gene expression was performed with clinical information from TCGA database. 566 robust DEGs (338 upregulated genes and 226 downregulated genes) were identified. Tumor tissue had a higher infiltration of resting dendritic cells and tumor-associated macrophages (TAM), including M0, M1, and M2 macrophages, while infiltration levels of B memory cells, plasma cells, T cells CD8, T follicular helper cells, and NK cells in normal tissue were relatively higher. GO terms and KEGG pathway analysis results revealed enrichment in tumor-associated pathways, including the extracellular matrix organization, cell−substrate adhesion cytokine−cytokine receptor interaction, calcium signaling pathway, and glycine, serine, and threonine metabolism, to name a few. Finally, FN1, MSLN, PLAU, and VCAN were selected as hub genes. High expression of FN1, MSLN, PLAU, and VCAN in PC significantly correlated with poor prognosis. Integrated transcriptomic analysis was used to provide new insights into PC pathogenesis. FN1, MSLN, PLAU, and VCAN may be considered as novel biomarkers of PC.

P-724 Gene expression profiling of cumulus cells from women with adenomyosis disclose differential pattern to endometriosis: striking the chord for evaluation of a biomarker

Abstract Study question Can metabolic picture of cumulus cell (CC) assess assisted reproductive technology outcome/s in women with adenomyosis/endometriosis and have implications on treatment decisions? Summary answer The differentially expressed genes between uterine pathologies explain the functional gap to identify diagnostic/prognostic markers of CC and further dissect molecular contribution to oocyte maturation. What is known already A healthy intra-follicular environment supports achievement of oocyte developmental competence (ODC) largely via coordinated CC-follicular fluid (FF) crosstalk. Lately, promising studies document CC of endometriosis are involved in important pathways for ODC like oxidative phosphorylation, metabolism, mitochondrial function and acetylation to name a few. Similar presentation/s of adenomyosis to other gynecological diseases including endometriosis makes the diagnosis difficult. Given that the functional integrities of CCs are susceptible to concurrent pathological conditions, understanding the influence of CCs on oocyte quality compared to routine morphological analysis in presence of deleterious systemic or pelvic conditions may impact clinical decisions. Study design, size, duration Twenty-eight consented patients undergoing IVF-ICSI between January to December 2021at Institute of Reproductive Medicine, Kolkata were enrolled. CCs obtained and corresponding oocyte maturation was assessed. They were sub-stratified (n = 9) according to uterine pathologies: adenomyosis (Group A), and endometriosis (Group B) as diagnosed by transvaginal ultrasound (TVUS). Counterpart/s of male sub-fertility, defined by World Health Organization criteria (5th edition) was treated as control (Group C; n = 10). CC from mature MII and immature GV oocytes were compared. Participants/materials, setting, methods CCs collected following oocyte-denudation were evaluated by Trypan Blue viability test. Patient age and embryo development were recorded with respect to each cumulus complex. Real Time-quantitative polymerase-chain-reaction (qRTPCR) was performed for glycolysis (PFKP, PKM2, ALDOA, LDHA, TPI1), cholesterol-biosynthesis (CYP51, MVK, PMVK, EBP) and extracellular-matrix (PTGS2, PTX3, TNFAIP6, VCAN) related gene/s known to have important functions in cumulus expansion and metabolism. Patient characteristics and gene expression were correlated by Spearman’s correlation. P &amp;lt; 0.05 was considered statistically significant. Main results and the role of chance A significant higher (p &amp;lt; 0.001) age (A vs B: 35.77±2.33 vs 25.88± 1.76; A vs C: 35.77±2.33 vs 31.9±1.98) and increased number of oocytes retrieved (A vs B: 11.22±1.56 vs 8.55±0.88) was documented in adenomyosis cohort. Immature oocytes from Group A expressed enhanced cholesterol/steroid biosynthesis, with significant (A vs B; p &amp;lt; 0.01) up-regulation of CYP51, MVK, PMVK, EBP in spite of hyper-estrogenic environment in peripheral blood. On contrary, significant attenuation (p &amp;lt; 0.01) of cholesterol biosynthesis was documented in endometriosis cohort compared to control. Expression of PTGS2, PTX3 and VCAN was down-regulated (p &amp;lt; 0.001) (which together mediate cumulus expansion and oocyte maturation, as well as corpus luteum formation and maintenance and reduce spindle abnormalities) in both Group A and B. Expression/s of CC-PFKP and PKM2 was lower in immature compared to mature oocytes (B vs C; p &amp;lt; 0.01) in endometriosis making the duo responsible for follicular/oocyte growth, and embryonic development. This supports the role in promoting cumulus expansion and oocyte competence. Expression levels of other genes including ALDOA, LDHA, TPI1, and TNFAIP6 were not associated with oocyte maturity, and fertilization in either cohort/s. Interestingly, levels of VCAN, TNFAIP6, PTX3 changed significantly as a function of the subject’s age and embryo development. Limitations, reasons for caution Our findings are limited by the relatively small sample size and inherent biological variability of human samples. Several yet known or unknown factors are presumably involved in the implantation process of adenomyosis and/or endometriosis. Multi-centric systematic studies are needed to corroborate these findings and account for the probable confounding factors. Wider implications of the findings A differential metabolic dependence of CC play essential role in oocyte competence in adenomyosis influencing cumulus expansion and oocyte maturation. This would be useful to do further studies to know how molecular interface of implantation capacity with live-birth rate would be affected to complement embryonic morphological assessment/s. Trial registration number Not Applicable

Greater expression of THBS2 and VACN in gastric carcinoma compared to gastritis.

e16026 Background: Gastric carcinoma (GC) is one of the most common cancers with the lowest survival rate in the world. The development from gastritis to GC may be related to genetic factors, but the specific molecular mechanism is not clear. The present study aimed to explore the gene difference between GC and gastritis. Methods: Two gene expression profiles including GC and gastritis were obtained from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) between GC and gastritis were explored. Through pathway enrichment of DEGs, hub genes were identified. The prognosis significances of hub genes for GC patients were analyzed by overall survival (OS) analysis. Animal models of gastritis and GC were constructed and immunofluorescence and RT-PCR were used to verify the distinct expression of hub genes between the two diseases. Finally, the correlations between hub genes and GC were understood based on BP neural network. Results: A total of 62 DEGs was identified between GC and gastritis. Eight hub genes were identified, including MSLN, FGA, SPP1, THBS2, SPARC, FN1, IGFBP7 and VCAN. Except SPP1, the high expression of the other seven hub genes predicted poor OS for GC patients. In mouse model, the mRNA expression of SPP1, FN1, IGFBP7, THBS2 and VCAN were significantly higher in GC group than that in gastritis group. No matter in mouse model or in Human Protein Atlas, the protein expression of THBS2 and VCAN in GC was higher than that in gastritis. Based on the strong correlation among THBS2, VACN and GC suggested by BP neural network, it was speculated that the expression levels of THBS2 and VACN may be a predictor of GC. Conclusions: Compared with gastritis, THBS2 and VACN are highly expressed in GC. THBS2 and VACN may be potential biomarkers and therapeutic targets for GC.

PCOS and Role of Cumulus Gene Expression in Assessing Oocytes Quality.

The global infertility rate has been declining from year to year. PCOS is one of the treatable accountable causes contributing to anovulatory infertility. Nevertheless, the success rate of treatments and live-birth outcomes especially involving assisted reproductive techniques is still not very promising. There is a reduction in the development potential of oocytes and high-quality embryos in PCOS patients compared to non-PCOS patients. A critical step in IVF treatment is the assessment of oocyte and embryo competence before embryo transfer. Oocytes in metaphase II are very fragile. Repeated morphological assessment on these oocytes may directly impair the quality and affect the whole process. Identification of potential biomarkers especially in the cumulus cells oocytes complex will help to predict the outcome and may create space for improvement. This review has explored gene expression in cumulus cells with regards to oocytes quality in both normal and PCOS women. The gene expression was classified according to their physiological function such as the contribution on cumulus expansion, cumulus cells apoptosis, and glucose metabolism. Collectively, the review suggested that positive expression of HAS2, PTX3, GREM1, and VCAN may correlate with good quality oocytes and can be used as an indicator among PCOS women.

Identification of novel therapeutic targets for Fuchs' endothelial corneal dystrophy based on gene bioinformatics analysis.

Fuchs’ endothelial corneal dystrophy (FECD) is a disease where progressive visual impairment occurs by the thickening of the Descemet’s membrane and the gradual degeneration and loss of corneal endothelial cells. This study aimed to investigate the key changes in gene expression associated with FECD and explore potential biomarkers and new therapeutic strategies for FECD. To explore the potential therapeutic targets of FECD, we downloaded the gene expression dataset GSE171830 from the Gene Expression Omnibus (GEO) database. A total of 303 differentially expressed genes (DEGs) were identified by the limma package. The enriched Gene Ontology (GO) annotations and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of DEGs mostly included the extracellular matrix organization, collagen-containing extracellular matrix, and the structural constituents of the extracellular matrix. Fifteen hub genes from the most significant module were ascertained by Cytoscape. Both collagen-containing extracellular matrix and extracellular matrix hit to ANXA1, VCAN, GPC3, TNC, IGFBP7, MATN3, and SPARCL1 genes in the GO cellular components. Among these genes, the expression of SPARCL1 was down-regulated in the FECD samples, whereas the expression of GPC3, MATN3, IGFBP7, TNC, VCAN, and ANXA1 was up-regulated in the FECD samples. Gene set enrichment analysis (GSEA) plots showed that among the 20,937 genes, SPARCL1 played an important role in three pathways, cytokine-cytokine receptor interaction, the TGF-beta signaling pathway, and antigen processing and presentation. The top three pathways enriched by the GPC3, MATN3, IGFBP7, TNC, VCAN, and ANXA1 genes were those for cytokine-cytokine receptor interaction, TGF-beta signaling, and RIG-I-like receptor signaling. In conclusion, the DEGs identified here might assist clinicians in understanding the pathogenesis of FECD. Furthermore, these identified biomarkers might serve as potential therapeutic targets for the treatment of FECD.

Protein source in maturation media affects gene expression in cumulus cells and embryo development in cattle

We aimed to evaluate if protein source (PS) alterations during IVM affect embryo sex/development and gene expression profile in cumulus cells (CCs). Bovine oocytes were matured and cultured in the presence of FBS or BSA. Then, the PS effect during IVM on gene expression (GPC4, VCAN, GHR, PTGS2, and ALCAM) was determined. CC biopsy was removed before and after IVM treatments. After fertilization and cultured, CCs were grouped according to their fate into CCs from immature COCs, CCs from COCs that did or did not result in embryos (according to PS). Results showed that when the culture was performed in FBS presence, blastocyst rate was higher (p < 0.05) than BSA. However, when embryos were cultured with BSA, no effect (p > 0.05) of PS during IVM was observed. PS used during IVM did not affect embryos sex (p > 0.05) but changed VCAN, GHR, PTGS2, and ALCAM genes expression. No differences (p > 0.05) were observed between immature and mature CCs groups in gene expression, regardless of their fate. Only the GHR gene was related to embryo production but just with FBS on IVM. In conclusion, PS can affect embryo development when using the serum on IVM and IVC, influences CCs gene expression, and has to be considered when studying oocyte quality markers.