The gene expression response of chronic lymphocytic leukemia cells to IL-4 is specific, depends on ZAP-70 status and is differentially affected by an NFκB inhibitor

PLoS One. 2014 Oct 3;9(10):e109533. doi: 10.1371/journal.pone.0109533. eCollection 2014.

Abstract

Interleukin 4 (IL-4), an essential mediator of B cell development, plays a role in survival of chronic lymphocytic leukemia (CLL) cells. To obtain new insights into the function of the IL-4 pathway in CLL, we analyzed the gene expression response to IL-4 in CLL and in normal B cells (NBC) by oligonucleotide microarrays, resulting in the identification of 232 non-redundant entities in CLL and 146 in NBC (95 common, 283 altogether), of which 189 were well-defined genes in CLL and 123 in NBC (83 common, 229 altogether) (p<0.05, 2-fold cut-off). To the best of our knowledge, most of them were novel IL-4 targets for CLL (98%), B cells of any source (83%), or any cell type (70%). Responses were significantly higher for 54 and 11 genes in CLL and NBC compared to each other, respectively. In CLL, ZAP-70 status had an impact on IL-4 response, since different sets of IL-4 targets correlated positively or negatively with baseline expression of ZAP-70. In addition, the NFκB inhibitor 6-Amino-4-(4-phenoxyphenethylamino)quinazoline, which reversed the anti-apoptotic effect of IL-4, preferentially blocked the response of genes positively correlated with ZAP-70 (e.g. CCR2, SUSD2), but enhanced the response of genes negatively correlated with ZAP-70 (e.g. AUH, BCL6, LY75, NFIL3). Dissection of the gene expression response to IL-4 in CLL and NBC contributes to the understanding of the anti-apoptotic response. Initial evidence of a connection between ZAP-70 and NFκB supports further exploration of targeting NFκB in the context of the assessment of inhibition of the IL-4 pathway as a therapeutic strategy in CLL, especially in patients expressing bad prognostic markers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis
  • Biomarkers, Tumor / genetics*
  • Blotting, Western
  • Case-Control Studies
  • Cell Proliferation
  • Cells, Cultured
  • Gene Expression Profiling*
  • Humans
  • I-kappa B Proteins / genetics*
  • Interleukin-4 / pharmacology*
  • Leukemia, Lymphocytic, Chronic, B-Cell / drug therapy
  • Leukemia, Lymphocytic, Chronic, B-Cell / genetics*
  • Leukemia, Lymphocytic, Chronic, B-Cell / pathology
  • Lymphocytes / cytology
  • Lymphocytes / metabolism
  • NF-kappa B / antagonists & inhibitors*
  • NF-kappa B / genetics
  • Oligonucleotide Array Sequence Analysis
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • ZAP-70 Protein-Tyrosine Kinase / genetics*

Substances

  • Biomarkers, Tumor
  • I-kappa B Proteins
  • IL4 protein, human
  • NF-kappa B
  • RNA, Messenger
  • Interleukin-4
  • ZAP-70 Protein-Tyrosine Kinase
  • ZAP70 protein, human

Grants and funding

This work was supported by grants from Plan Nacional de I+D+I 2004-2007, Programa de Promoción de la Investigación Biomédica y en Ciencias de la Salud del Ministerio de Sanidad y Consumo (PI07/0135 to AP) and Plan Nacional de I+D+I 2008-2011, Acción Estratégica en Salud (PI10/01226 to AP) (Instituto de Salud Carlos III (ISCIII) co-financed with European Regional Development Funds, “Una manera de hacer Europa”), II PCTRM 2007–2010, Fundación Séneca, Agencia de Ciencia y Tecnología de la Región de Murcia (08721/PI/08 to AP, and 04487/GERM/06 to MRAL), ISCIII Spanish Cell Therapy Network (Tercel; RD06/0010/0023 to JMM), and the CIBERehd scientific program to MRAL. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.