Treatment of chronic lymphocytic leukemia cells with signaling inhibitors. Treatment of chronic lymphocytic leukemia cells with signaling inhibitors

To identify the molecular targets of treatment, we studied gene expression profile of primary chronic lymphocytic leukemia (CLL) cells isolated from the peripheral blood of CLL patients and treated in vitro with two distinct signaling inhibitors: 1) Dimethyl fumarate targeting inflammatory pathways 2) VIVIT peptide targeting NFAT transcription factor activation. Overall design: CLL B-cells were negatively selected and purified from patient’s total peripheral blood mononuclear cells (PBMC), cultured in RPMI with 10% heat inactivated FBS and Pen-Strep and stimulated with DMF 100 µM or VIVIT-11R 10µM for 4 hours and RNAseq was performed. Untreated samples were used as controls.

为明确治疗相关的分子靶点，本研究对从慢性淋巴细胞白血病（chronic lymphocytic leukemia, CLL）患者外周血中分离的原代CLL细胞开展基因表达谱分析。上述细胞经两种不同的信号通路抑制剂体外处理：1）靶向炎症通路的富马酸二甲酯（Dimethyl fumarate）；2）靶向NFAT转录因子（NFAT transcription factor）激活的VIVIT肽（VIVIT peptide）。
整体实验设计：从患者外周血单个核细胞（peripheral blood mononuclear cells, PBMC）中通过阴性筛选法纯化CLL B细胞，将其培养于添加10%热灭活胎牛血清（FBS）及青霉素-链霉素双抗（Pen-Strep）的RPMI培养基中，分别以100 µM富马酸二甲酯（DMF）与10 µM VIVIT-11R处理4小时，随后进行RNA测序（RNAseq）。以未接受处理的样本作为对照。