FIGURE 1.

Ibrutinib modifies NLC gene expression profile. NLC were generated from PBMCs of 10 CLL patients after 15 days of culture. Then, CLL cells were carefully washed off by vigorously pipetting and adherent NLC were treated over-night with 1 μM ibrutinib or vehicle (DMSO). (A) Heat map depicts differentially expressed genes between NLC treated and not with ibrutinib. Genes were defined as differentially expressed between ibrutinib-treated vs. vehicle-treated group at a significant level of P < 0.05 and with a fold change cut off ±2. The supervised analysis identified 566 differentially expressed genes, 409 down-regulated and 157 up-regulated by treatment. (B) Among down-regulated genes, the most represented GO categories were related to immune system process, inflammatory response, immune response, cytokine activity, implying the ability of ibrutinib to modify the expression of genes implicated in immune function of NLC. (C) The down-regulated profile included several genes belonging to interleukin 1 (IL-1β) and tumor necrosis factor receptor family (TNF-α). Moreover, we found the down-regulation of several chemokine CXCL12 and CSF1. Values of untreated and treated samples (n = 10) are connected by lines (**P < 0.01). (D) Bar diagram depicts the level of chemokines and interleukins secreted by NLC either treated or not with ibrutinib. Secretion was measured on NLC supernatants by ELISA (n = 20, *P < 0.05, **P < 0.01).