Project description:NOS1 plays a vital role in tumor. A cell model of NOS1 gene knockout in human melanoma cell line A375 was constructed using CRISPR/Cas9 technique for the study of its function. We used microarrays to detail the Global gene expression underlying NOS1-knockout compare to wild type A375 cell.
Project description:The experiment aimed to study mechanisms of melanoma drug resistance. ARID1A confers resistance to trametinib and therefore RNAseq was collected alongside other omics data for parental A375 likes, and A375 lines with ARID1A KO, in the presence of no drug, trametinib, vemurafenib or their combination
Project description:Endogenous nitric oxide (NO) produced by nitric oxide synthases (NOSs) plays an important immunosuppressive role in the tumor microenvironment. In melanoma, NOS1 expression was increased with tumor progression and correlated with tumor immune escape through inhibition on type I interferon (IFN) signaling. However, the immune regulatory role and its related mechanism of NOS1 and its impacts on immune therapy such as immune checkpoint blockade (ICB) in melanoma is unclear. Here, we found that NOS1 expression induced IRF7 modification by s-nitrosylation at C435 site in mouse (C481 in human), which functionally promotes tumor growth in mouse models. Mechanically, IRF7-C435-SNO inhibited IFNβ transcription under PRR signal activation, leading to disorder in initiation of type I interferons response in melanoma cell. In melanoma mouse model, IRF7-C435-SNO decreased infiltration and activation of CD8 T cells in tumor microenvironment, by reducing antigen presentation processes in tumor cells and inhibits the maturation of DC1. Clinically, high expression of NOS1 correlated with poor survival prognosis and resistance with to ICB anti-tumor therapies in melanoma cases with less immune cell infiltration. Our study suggested that NOS1 expression in melanoma characterizes IFN-I signal disorders in response to innate immune stimulation by IRF7 s-nitrosylation. Targeting NOS1 signaling might benefit for overcome of immune therapeutically resistance particularly in immune cold melanoma phenotype.
Project description:BRAF-inhibitor (BRAFi)-resistance compromises long term survivorship of malignant melanoma patients, and mutant NRAS is a major mediator of BRAFi-resistance. We have employed NanoString nCounterTM transcriptomic analysis of isogenic human malignant melanoma cells that differ only by NRAS mutational status (BRAFi-sensitive A375-BRAFV600E/NRASQ61 versus BRAFi-resistant A375-BRAFV600E/NRASQ61K), identifying modulation of specific gene expression networks as a function of NRASQ61K-status.
Project description:Endogenous nitric oxide (NO) produced by nitric oxide synthases (NOSs) plays an important immunosuppressive role in the tumor microenvironment. In melanoma, NOS1 expression was increased with tumor progression and correlated with tumor immune escape through inhibition on type I interferon (IFN) signaling. However, the immune regulatory role and its related mechanism of NOS1 and its impacts on immune therapy such as immune checkpoint blockade (ICB) in melanoma is unclear. Here, we found that NOS1 expression induced IRF7 modification by s-nitrosylation at C435 site in mouse (C481 in human), which functionally promotes tumor growth in mouse models. Mechanically, IRF7-C435-SNO inhibited IFNβ transcription under PRR signal activation, leading to disorder in initiation of type I interferons response in melanoma cell. In melanoma mouse model, IRF7-C435-SNO decreased infiltration and activation of CD8 T cells in tumor microenvironment, by reducing antigen presentation processes in tumor cells and inhibits the maturation of DC1. Clinically, high expression of NOS1 correlated with poor survival prognosis and resistance with to ICB anti-tumor therapies in melanoma cases with less immune cell infiltration. Our study suggested that NOS1 expression in melanoma characterizes IFN-I signal disorders in response to innate immune stimulation by IRF7 s-nitrosylation. Targeting NOS1 signaling might benefit for overcome of immune therapeutically resistance particularly in immune cold melanoma phenotype.
Project description:Identify transcriptionnally and translationally regulated mRNA in melanoma parental and persister cells In this dataset, we include expression data of A375 melanoma drug-naïve parental cells and A375 melanoma persister cells that survived from BRAF and MEK inhibition. The expression data are studied in both total RNA and polysome-bounded RNA.
Project description:Human melanoma cell line A375 and A2058 were incubated with DMSO (control group) or Vemurafenib (1 μM in DMSO) for 72 hours. Total RNA was isolated with Qiagen RNA isolation kit; 1.5 μg of total RNA was used as template for cDNA synthesis.