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Transcriptomic analysis of E0771 tumors treated with FEC + oHSV-1 Vs PBS

ABSTRACT: Transcriptomic analysis of E0771 tumors treated with FEC + oHSV-1 Vs PBS

PROVIDER: PRJNA762241 | ENA |

REPOSITORIES: ENA

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Transcriptomic analysis of E0771 tumors treated with FEC + oHSV-1 Vs PBS

Project description:E0771 tumors were implanted either subcutaneously on the left flank or orthotopically in the mammary fat pad and treated with either PBS or FEC + oHSV-1 Mice were sacrificed 5 days after the start of treatment and whole tumour digests were used for RNA extraction

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FEC + OV treatment of TNBC tumors

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RNA-seq of E0771-Vector control, E0771-Malt1-WT and E0771 Malt1-PD after T cell killing

Project description:To understand potential downstream signaling molecules that are responsible for wild type-Malt1 induced resistance of tumor cell to CD8 T cell killing , we first use E0771 cells expressing either -Vector control, wild type-Malt1 (M-WT), or Malt1 PD mutant (M-PD) to coculture with activated CD8 T cell at a effctor: target ratio(E:T ration) of 3 for 12 hours. Then wash the cells with PBS, digest the cells by 0.25% trypsin solution, harvest the cells in 15 ml tubes by centrifugation (1000 rpm 5min) and seed cells into the original plate with DMEM+10%FBS+1%P/S medium. 6 hours after sseding, wash cells with PBS, lyse cells with TRIZOL for RNA extraction, perform reverse transcription and cDNA library for RNA-Seq analysis.

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IDH status dictates oHSV mediated metabolic reprogramming affecting anti-tumor immunity

Project description:Identification of IDH mutations has uncovered the crucial role played by metabolism in glioma-genesis. Oncolytic herpes virus (oHSV) therapy initiates direct tumor debulking by tumor lysis and also activates antitumor immunity however little is known about the role of glioma metabolism in determining oHSV efficacy. Here we identified that oHSV therapy rewired central carbon metabolism with increased glucose utilization towards oxidative phosphorylation and shuttled glutamine towards reductive carboxylation in IDH wildtype (wt) glioma. The switch in metabolism resulted in increased lipid synthesis, and cellular ROS. PKC induced ACSL4 in oHSV treated cells led to lipid peroxidation and ferroptosis. Ferroptosis was critical to launch an antitumor immune response important for efficacy. Mutant IDH (IDHR132H) gliomas are incapable of reductive carboxylation and hence ferroptosis. Pharmacological blockade of IDHR132H induced ferroptosis and antitumor immunity. This study provides a rationale to treat high grade IDHR132H glioma patients under oHSV treatment with IDHR132H inhibitor.

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Gene expression profile at single cell level of CD45+ immune cells from E0771-Vector control tumor tissue, E0771 wildtype Malt1 overexpression tumor tissue and E0771 paracaspase-deficient Malt1 overexpression tumor tissue.

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RNA-seq of E0771-Vector control, E0771-Malt1-WT and E0771 Malt1-PD (Without T cell coculture)

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