PIM kinases are essential for chronic lymphocytic leukemia cell survival (PIM2/3) and CXCR4-mediated microenvironmental interactions (PIM1)

Mol Cancer Ther. 2014 May;13(5):1231-45. doi: 10.1158/1535-7163.MCT-13-0575-T. Epub 2014 Mar 21.

Abstract

Overexpression of the CXCR4 receptor is a hallmark of chronic lymphocytic leukemia (CLL) and is important for CLL cell survival, migration, and interaction with their protective microenvironment. In acute myelogenous leukemia (AML), PIM1 was shown to regulate the surface expression of the CXCR4 receptor. Here, we show that PIM (proviral integration site for Moloney murine leukemia virus) kinases 1-3 are overexpressed and that the CXCR4 receptor is hyperphosphorylated on Ser339 in CLL compared with normal lymphocytes. Furthermore, CXCR4 phosphorylation correlates with PIM1 protein expression and PIM1 transcript levels in CLL. PIM kinase inhibition with three different PIM kinase inhibitors induced apoptosis in CLL cells independent of the presence of protective stromal cells. In addition, PIM inhibition caused dephosphorylation of the CXCR4 receptor on Ser339, resulting in enhanced ligand-dependent CXCR4 internalization and reduced re-externalization after withdrawal of CXCL12. Furthermore, PIM inhibition in CLL cells blocked CXCR4 functions, such as migration toward CXCL12- or CXCL12-induced extracellular signal-regulated kinase (ERK) phosphorylation. In concordance, pretreatment of CLL cells with PIM kinase inhibitors strongly reduced homing of CLL cells toward the bone marrow and the spleen of Rag2(-/-)γc(-/-) mice in vivo. Interestingly, the knockdown of PIM kinases in CLL cells demonstrated diverging functions, with PIM1 regulating CXCR4 surface expression and PIM2 and PIM3 as important for the survival of CLL cells. Our results show that PIM kinase inhibitors are an effective therapeutic option for CLL, not only by impairing PIM2/3-mediated CLL cell survival, but also by blocking the PIM1/CXCR4-mediated interaction of CLL cells with their protective microenvironment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Bone Marrow / metabolism
  • Cell Line, Tumor
  • Cell Membrane / metabolism
  • Cell Movement / drug effects
  • DNA-Binding Proteins / deficiency
  • DNA-Binding Proteins / genetics
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Gene Expression
  • Gene Knockdown Techniques
  • Humans
  • Leukemia, Lymphocytic, Chronic, B-Cell / metabolism*
  • Leukemia, Lymphocytic, Chronic, B-Cell / pathology
  • Mice
  • Mice, Knockout
  • Nuclear Proteins / deficiency
  • Nuclear Proteins / genetics
  • Phosphorylation
  • Protein Binding
  • Protein Kinase Inhibitors / pharmacology
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / metabolism
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-pim-1 / antagonists & inhibitors
  • Proto-Oncogene Proteins c-pim-1 / genetics
  • Proto-Oncogene Proteins c-pim-1 / metabolism*
  • RNA, Small Interfering / genetics
  • Receptors, CXCR4 / metabolism*
  • Spleen / metabolism
  • Stromal Cells / drug effects
  • Stromal Cells / metabolism
  • Tumor Microenvironment

Substances

  • DNA-Binding Proteins
  • Nuclear Proteins
  • PIM2 protein, human
  • Protein Kinase Inhibitors
  • Proto-Oncogene Proteins
  • RAG2 protein, human
  • RNA, Small Interfering
  • Receptors, CXCR4
  • Protein-Serine-Threonine Kinases
  • Proto-Oncogene Proteins c-pim-1
  • proto-oncogene proteins pim
  • Extracellular Signal-Regulated MAP Kinases