Project description:Identification of transcriptional program influenced by the expression of lncRNA linc00941 in malignant pleural mesothelioma cell line MSTO-211H. Linc00941 expression was knocked-down through siRNA strategy.
Project description:To investigate the effect of afuresertib on gene expression, we have employed whole genome microarray expression profiling as a discovery platform to identify genes with the potential to suppress the cell proliferation of MPM cell lines. MSTO-211H and ACC-MESO-4 cells were treated with afuresertib (10 μmol/L) for 24 h in vitro.
Project description:Immunoaffinity purification was performed on human mesothelioma cell lines NCI-H2452, NCI-H28, MSTO-211H and JL1, on murine mesothelioma cell line AB12, as well as on mesothelioma samples from two patients (including tumor and benign tissues). Thereafter Immunopeptidomics by Mass Spectrometry on a Tims TOF Pro revealed the MHC peptide landscape of mesothelioma.
Project description:The goal of this experiment was to get deep into TRIM28 biological function in malignant pleural mesothelioma. To this end MSTO-211H cell line was infected by two different sgRNAs targeting TRIM28 and a non-targeting sgRNA as control. Two independent experiments were performed.RNA was collected 7 days after infection and changes in gene expression were analyzed by mRNA-seq.
Project description:Conditional expression of sh-YAP1 modulates YAP1/TEAD-dependent transcription and causes regression of established human malignant mesothelioma MSTO-211H [sh-YAP1] xenografts.
Project description:GPC2 and GD2 are validated CAR T cell targets in neuroblastoma, but durable responses remain limited. We profiled the surfaceome of neuroblastoma-derived extracellular vesicles (EVs) and assessed their impact on CAR T cell function. Neuroblastoma EVs displayed GPC2 and GD2, with minimal PD-L1, and were detected in blood from tumor-bearing mice and patients. These EVs directly activated paired CAR T cells, suggesting a role for a peripheral source of CAR antigen. To exploit this therapeutically, we engineered non-tumor-derived GPC2+ synthetic EVs (SyntEVs) as CAR T cell enhancers and armored them with either albumin- or GD2-binding domains. In mice harboring human neuroblastomas, serial infusion of armored SyntEVs following GPC2 CAR T cells enhanced tumor control by boosting peripheral CAR T cell persistence. Moreover, GD2-targeting SyntEVs decorated low-antigen tumor cells with GPC2, circumventing antigen downregulation. This SyntEV platform offers a versatile system to address the therapeutic limitations of CAR T cells in solid tumors.
Project description:Chimeric antigen receptor (CAR) T cells have revolutionized the landscape of cancer therapy, demonstrating unprecedented success in treating relapsed or refractory blood cancers. Previous studies of the mechanisms underlying the interactions and responses of CAR T cells and their targets have focused on the activation of CAR T cells and attempted to optimize CAR design to increase efficacy, while ignoring tumors and their responses to CAR ligation. We evaluated the signaling of a second-generation, ligand-based CAR built from the colony-stimulating factor 1 (CSF1) ligand to target the CSF1 receptor (CSF1R) on CSF1R-expressing target cells, and compared it to a conventional single-chain variable fragment (scFv)–based CAR against the B-cell antigen CD19 using stable isotope labeling by amino acids in cell culture (SILAC) co-culture with phosphotyrosine (pY) enrichment and liquid chromatography–tandem mass spectrometry (LC–MS/MS).. We showed that ligation of CSF1R-expressing THP-1 cells with CSF1R-CAR T cells stimulated CSF1R-like signaling in the THP-1 cells, whereas no target cell signaling response was observed after the ligation of CD19-CAR T cells with target Raji cells. In experiments with small-molecule inhibitors of the tyrosine kinase Lck, actin polymerization, and CSF1R, we found that CAR-induced CSF1R signaling in THP-1 cells depended exclusively on the kinase activity of CSF1R with no participation from T cell activation. Consistently, CSF1R-CAR T cells promoted THP-1 cell growth at low effector-to-target (E:T) ratios but prevented THP-1 cell growth at high E:T ratios. Our data provide evidence for an unintended consequence of CARs: CAR-induced signaling in cancer cells. These data may have broad implications for the choice of CAR antigen for optimal clinical efficacy.