Integrated analysis reveals FLI1 regulates the tumor immune microenvironment via its cell-type-specific expression and transcriptional regulation of distinct target genes of immune cells in breast cancer.

BACKGROUND: Immunotherapy is a practical therapeutic approach in breast cancer (BRCA), and the role of FLI1 in immune regulation has gradually been unveiled. However, the specific role of FLI1 in BRCA was conflicted; thus, additional convincing evidence is needed. METHODS: We explored the upstream regulation of FLI1 expression via summary data-based Mendelian randomization (SMR) analysis and ncRNA network construction centering on FLI1 using BRCA genome-wide association study (GWAS) summary data with expression quantitative trait loci (eQTLs) and DNA methylation quantitative trait loci (mQTLs) from the blood and a series of in silico analyses, respectively. We illuminated the downstream function of FLI1 in immune regulation by integrating a series of analyses of single-cell RNA sequence data (scRNA-seq). RESULTS: We verified a causal pathway from FLI1 methylation to FLI1 gene expression to BRCA onset and demonstrated that FLI1 was downregulated in BRCA. FLI1, a transcription factor, served as myeloid and T cells' communication regulator by targeting immune-related ligands and receptor transcription in BRCA tissues. We constructed a ceRNA network centering on FLI1 that consisted of three LncRNAs (CKMT2-AS1, PSMA3-AS1, and DIO3OS) and a miRNA (hsa-miR-324-5p), and the expression of FLI1 was positively related to a series of immune-related markers, including immune cell infiltration, biomarkers of immune cells, and immune checkpoints. CONCLUSION: Low-methylation-induced or ncRNA-mediated downregulation of FLI1 is associated with poor prognosis, and FLI1 might regulate the tumor immune microenvironment via a cell-type-specific target genes manner in BRCA.

Whole-exome sequencing identified a homozygous novel RAG1 mutation in a child with omenn syndrome.

INTRODUCTION AND OBJECTIVES: Omenn syndrome (OS) is a very rare type of severe combined immunodeficiencies manifested with erythroderma, eosinophilia, hepatosplenomegaly, lymph-adenopathy, and elevated level of serum IgE. OS is inherited with an autosomal recessive mode of inheritance. Germline mutations in the human RAG1 gene cause OS. MATERIALS AND METHODS: In this study, we investigated a 2-month-old boy with cough, mild anaemia, pneumonia, immunodeficiency, repeated infection, feeding difficulties, hepatomegaly, growth retardation, and heart failure. Parents of the proband were phenotypically normal. RESULTS: Karyotype analysis and chromosomal microarray analysis found no chromosomal structural abnormalities (46, XY) and no pathogenic copy number variations (CNVs) in the proband. Whole-exome sequencing identified a novel homozygous single nucleotide deletion (c.2662delC) in exon 2 of the RAG1 gene in the proband. Sanger sequencing confirmed that both the proband parents were carrying this variant in a heterozygous state. This variant was not identified in two elder sisters and one elder brother of the proband and in the 100 ethnically matched normal healthy individuals. This novel homozygous deletion (c.2662delC) leads to the frameshift, which finally results in the formation of the truncated protein (p.Leu888Phefs*3) V(D)J recombination-activating protein 1 with 890 amino acids compared with the wildtype V(D)J recombination-activating protein 1 of 1043 amino acids. Hence, it is a loss-of-function variant. CONCLUSIONS: Our present study expands the mutational spectrum of the RAG1 gene associated with OS. We also strongly suggested the importance of whole-exome sequencing for the genetic screening of patients with OS.

Identification of key genes controlling breast cancer stem cell characteristics via stemness indices analysis.

BACKGROUND: With the gradual unveiling of tumour heterogeneity, cancer stem cells (CSCs) are now being considered the initial component of tumour initiation. However, the mechanisms of the growth and maintenance of breast cancer (BRCA) stem cells are still unknown. METHODS: To explore the crucial genes modulating BRCA stemness characteristics, we combined the gene expression value and mRNA expression-based stemness index (mRNAsi) of samples from The Cancer Genome Atlas (TCGA), and the mRNAsi was corrected using the tumour purity (corrected mRNAsi). mRNAsi and corrected mRNAsi were analysed and showed a close relationship with BRCA clinical characteristics, including tumour depth, pathological staging and survival status. Next, weighted gene co-expression network analysis (WGCNA) was applied to distinguish crucial gene modules and key genes. A series of functional analyses and expression validation of key genes were conducted using multiple databases, including Oncomine, Gene Expression Omnibus (GEO) and Gene Expression Profiling Integrative Analysis (GEPIA). RESULTS: This study found that mRNAsi and corrected mRNAsi scores were higher in BRCA tissues than that in normal tissues, and both of them increased with tumour stage. Higher corrected mRNAsi scores showed worse overall survival outcomes. We screened 3 modules and 32 key genes, and those key genes were found to be strongly correlated with each other. Functional analysis revealed that the key genes were related to cell fate decision events such as the cell cycle, cellular senescence, chromosome segregation and mitotic nuclear division. Among 32 key genes, we identified 12 genes that strongly correlated with BRCA survival. CONCLUSIONS: Thirty-two genes were found to be closely related to BRCA stem cell characteristics; among them, 12 genes showed prognosis-oriented effects in BRCA patients. The most significant signalling pathway related to stemness in BRCA was the cell cycle pathway, which may support new ideas for screening therapeutic targets to inhibit BRCA stem characteristics. These findings may highlight some therapeutic targets for inhibiting BRCA stem cells.

Curcumin induced human gastric cancer BGC-823 cells apoptosis by ROS-mediated ASK1-MKK4-JNK stress signaling pathway.

The signaling mediated by stress-activated MAP kinases (MAPK), c-Jun N-terminal kinase (JNK) has well-established importance in cancer. In the present report, we investigated the effects of curcumin on the signaling pathway in human gastric cancer BGC-823 cells. Curcumin induced reactive oxygen species (ROS) production and BGC-823 cells apoptosis. Inhibition of ROS generation by antioxidant (NAC or Trion) significantly prevented curcumin-mediated apoptosis. Notably, we observed that curcumin activated ASK1, a MAPKKK that is oxidative stress sensitive and responsible to phosphorylation of JNK via triggering cascades, up-regulated an upstream effector of the JNK, MKK4, and phosphorylated JNK protein expression in BGC-823 cells. However, curcumin induced ASK1-MKK4-JNK signaling was attenuated by NAC. All the findings confirm the possibility that oxidative stress-activated ASK1-MKK4-JNK signaling cascade promotes the apoptotic response in curcumin-treated BGC-823 cells.

The causal effects of genetically determined immune cells on gynecologic malignancies: a Mendelian randomization study.

Background: Evidence from observational studies suggested a connection between immune cells and gynecologic malignancies. To investigate potential causative associations between immunophenotype traits and gynecologic malignancies, we used a two-sample Mendelian randomization analysis. Methods: The genetic instrumental variables of 731 immunophenotypes of peripheral blood were obtained by the GWAS database; the GWAS data of common gynecologic cancers were obtained from FinnGen study. The main statistic method was the inverse-variance weighted method. We also used the weighted mode, weighted median, and MR Egger for evaluations. The MR Steiger directionality test was further used to ascertain the reverse causal relationship between immune cells and gynecologic cancers. Results: We identified 50 highly probable immunophenotypes and 65 possible ones associated with gynecologic malignancies. The majority of the B cell panel was protective factors in cervical cancer. However, there was a correlation found in the B cells panel with a probable factor associated with an elevated risk of endometrial cancer. Immunophenotypes in the monocyte panel were linked to a lower probability of ovarian cancer and vulvar cancer. All of the gynecologic cancers in our study had no statistically significant impact on immune cells, according to reverse MR analysis. Conclusion: Our study firstly emphasized the genetically predicted causality between immune cells and gynecologic malignancies. This knowledge will be critical to formulating the measures to prevent malignancies in female at risk in future clinical practice.

Construction and characterization of a cuproptosis- and immune checkpoint-based LncRNAs signature for breast cancer risk stratification.

BACKGROUND: Cuproptosis is the most recently identified form of cell death, and copper homeostasis is an important cancer therapeutic target. However, the therapeutic benefits of cuproptosis-targeted treatment in BRCA remain undetermined. This study utilized LncRNAs linked to cuproptosis genes and immune checkpoint genes to generate a BRCA predictive signature. METHODS: We screened a population of LncRNAs that correlated with both cuproptosis genes and immune checkpoint genes and used ten of these LncRNAs to construct a prognosis-predictive signature. We then validated and proved the efficacy of the signature in predicting the prognosis of BRCA patients. We also unraveled the relationship between the signature and the immunological milieu, immune function, and susceptibility to chemotherapy. RESULTS: The signature derived from the ten cuproptosis- and immune-related prognostic LncRNAs (CuImP-LncRNAs) can be implied to categorize patients into two groups, including the high- and low-risk groups. The value of the signature was validated, and the risk score was verified as an independent prognostic indicator. The TIME and TMB distribution patterns and chemosensitivity were depicted in the high- and low-risk groups, respectively. Patients of the high-risk group with a suppressive immunological intratumor context were more sensitive to a broad range of antitumor agents. In contrast, low-risk individuals with active immune function responded more favorably to immunotherapy. CONCLUSION: Our findings provided a novel and effective model for predicting BRCA prognosis and the propensity to different treatment modalities, thus contributing to the optimization of personalized BRCA therapy in the future.

CD40LG as a Prognostic Molecular Marker Regulates Tumor Microenvironment Through Immune Process in Breast Cancer.

PURPOSE: Breast cancer (BRCA) is the second most common malignancy in the world and the most common in women. Here, we utilized publicly available BRCA dataset to investigate potential prognosis-related genes through integrated bioinformatics analysis. MATERIALS AND METHODS: BRCA dataset was obtained from the Cancer Genome Atlas (TCGA) database. The ESTIMATE algorithm was used to calculate the ImmuneScores and StromalScores of the samples and then divided them into high- and low-score groups based on the median score. Common differentially expressed genes (DEGs) were identified through differential expression analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. The core prognostic genes were the intersection of hub genes from PPI network and prognostic genes from univariate Cox proportional hazard regression analysis. Finally, the CIBERSORT algorithm was used to calculate proportions of 22 tumor-infiltrating immune cells (TICs) in BRCA samples. RESULTS: A total of 486 DEGs were identified. These genes were mainly enriched in immune-related pathways. Crossover genes between the hub genes and the prognostic genes were CD2 and CD40LG. CD40LG was further investigated in this study. CD40LG was downregulated in BRCA samples compared with normal samples, and a lower CD40LG expression was associated with advanced tumor stages and a poor prognosis. CD40LG was shown to be involved in immune-related pathways of BRCA by Gene Set Enrichment Analysis. Finally, 14 TICs were found to have a close relationship with CD40LG. CONCLUSION: CD40LG was found to be a core prognostic gene related to tumor microenvironment and deeply involved in immune-related pathways in BRCA. Our findings may provide new insights for exploring the molecular mechanisms of BRCA and developing new immunotherapies for the disease.

Docetaxel-loaded ultrasmall nanostructured lipid carriers for cancer therapy: in vitro and in vivo evaluation.

Lack of cancer-targeted delivery of chemotherapeutics is one of the major obstacles for successful cancer therapy. Nanostructured lipid carriers (NLC) have shown great promise in drug-delivery applications since they are highly scalable, biodegradable nanocarriers with high-drug-loading capacity. However, traditional method prepared NLC, the diameter of which range from 80 to 200 nm, is easily blocked and trapped in perivascular regions without further penetration. As a result, ultrasmall NLC with size under 100 nm or lower range are reported to be ideally tumor targeting carrier as it allows for superior tumor accumulation and permeation. Moreover, surface modification of NLC with folic acid (FA) could significantly increase the drug-delivery efficiency through active targeting effect. In our study, an ultrasmall NLC with FA modification (FA-NLC) was prepared to load docetaxel (DTX) for cancer therapy. Our results showed that DTX-loaded FA-NLC comprised of homogeneous particles with size around 30 nm. In addition, it exhibited great colloidal stability, satisfactory drug-loading efficiency, and high biocompatibility in vitro. Meanwhile, in vivo studies indicated that ultrasmall FA-NLC exhibited greater tumor retention and enhanced antitumor effect compared with control.

Identification and Validation of an Immunological Expression-Based Prognostic Signature in Breast Cancer.

Background: Emerging evidence suggests that the immune system plays a crucial role in the regulation of the response to therapy and long-term outcomes of patients with breast cancer (BRCA). In this study, we aimed to identify a significant signature based on immune-related genes to predict the prognosis of BRCA patients. Methods: The expression data were downloaded from The Cancer Genome Atlas (TCGA). The immune-related gene list, the transcription factor (TF) gene list, and the immune infiltrate scores of samples in the TCGA database were acquired from the ImmPort database, the Cistrome Cancer database, and the TIMER database, respectively. Univariate Cox regression analysis was utilized to identify prognostic immune-related differentially expressed genes (DEGs) (PIRDEGs) in BRCA. A prognostic immune signature containing 15 PIRDEGs in BRCA was established using the least absolute shrinkage and selection operator (LASSO) model with 1,000 iterations followed by a stepwise Cox proportional hazards model with a training set of 508 samples in TCGA. An independent assessment of the prognostic prediction ability of the signature was conducted using Kaplan-Meier survival analysis with a testing set of 505 samples in TCGA. Results: We identified 466 PIRDEGs and 80 TFs among the DEGs. A gene signature containing 15 PIRDEGs was constructed. Risk scores of BRCA patients were calculated using this model, which showed a high accuracy of prognosis prediction in both the training set and testing set and could be an independent prognostic factor of BRCA patients. Conclusions: Our study revealed that a PIRDEG signature could be a candidate prognostic biomarker for predicting the overall survival (OS) of patients with BRCA.

EMIMBF4-Assisted SnO2-Based Planar Perovskite Films for Label-Free Photoelectrochemical Sensing.

In this work, a photoelectrochemical (PEC) sensing system was developed for chemical detection without any tags. The system is based on methylammonium lead iodide chloride perovskite as the photoelectrochemical signal-generating molecule, tin oxide nanoparticles (SnO2) as the electron transport layer, and ionic liquid of 1-ethyl-3-methylimidazolium tetrafluoroborate (EMIMBF4) as the stabilizer and electrolyte. The photosensitizer film is formed through spin-coating followed by low-temperature sintering. Upon irradiation with 473 nm light, an anodic photocurrent is generated in a blank electrolyte and changed significantly after addition of hemin into the electrolyte. The change of photocurrent after addition of hemin suggested that the chemicals participated in a photoelectrochemical process. Our work paves the way to developing analytical methods with perovskite.

Synthesis, anti-HBV activity and renal cell toxicity evaluation of mixed phosphonate prodrugs of adefovir.

A series of phosphonate ester prodrugs of adefovir incorporating l-amino (thio)acid and non-steroidal anti-inflammatory drug (NSAID) moieties were synthesized and their anti-HBV activity and renal cell toxicity were evaluated in HepG2 2.2.15 and HK-2 cells respectively. Bioactivity evaluation results revealed that this kind of adefovir prodrug have lower renal cell toxicity than adefovir dipivoxil. Compounds 8a and 8b, incorporating the NSAID ketoprofen and the l-amino acid (Val or Ile) structural fragments, exhibited more potent anti-HBV activity than adefovir dipivoxil with IC(50) = 0.51 and 0.73 µM, SI = 1697.64 and 881.92 respectively. In vitro stability studies showed that the synthesized prodrugs have higher chemical and plasma stability than the positive control adefovir dipivoxil.