Inhibition of NF-κB-mediated signaling by the cyclin-dependent kinase inhibitor CR8 overcomes prosurvival stimuli to induce apoptosis in chronic lymphocytic leukemia cells

Clin Cancer Res. 2013 May 1;19(9):2393-405. doi: 10.1158/1078-0432.CCR-12-2170. Epub 2013 Mar 26.

Abstract

Purpose: Chronic lymphocytic leukemia (CLL) is currently incurable with standard chemotherapeutic agents, highlighting the need for novel therapies. Overcoming proliferative and cytoprotective signals generated within the microenvironment of lymphoid organs is essential for limiting CLL progression and ultimately developing a cure.

Experimental design: We assessed the potency of cyclin-dependent kinase (CDK) inhibitor CR8, a roscovitine analog, to induce apoptosis in primary CLL from distinct prognostic subsets using flow cytometry-based assays. CLL cells were cultured in in vitro prosurvival and proproliferative conditions to mimic microenvironmental signals in the lymphoid organs, to elucidate the mechanism of action of CR8 in quiescent and proliferating CLL cells using flow cytometry, Western blotting, and quantitative real-time PCR.

Results: CR8 was 100-fold more potent at inducing apoptosis in primary CLL cells than roscovitine, both in isolated culture and stromal-coculture conditions. Importantly, CR8 induced apoptosis in CD40-ligated CLL cells and preferentially targeted actively proliferating cells within these cultures. CR8 treatment induced downregulation of the antiapoptotic proteins Mcl-1 and XIAP, through inhibition of RNA polymerase II, and inhibition of NF-κB signaling at the transcriptional level and through inhibition of the inhibitor of IκB kinase (IKK) complex, resulting in stabilization of IκBα expression.

Conclusions: CR8 is a potent CDK inhibitor that subverts pivotal prosurvival and proproliferative signals present in the tumor microenvironment of CLL patient lymphoid organs. Our data support the clinical development of selective CDK inhibitors as novel therapies for CLL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Antineoplastic Agents / pharmacology*
  • Apoptosis*
  • CD40 Ligand / physiology
  • Cell Cycle Checkpoints / drug effects
  • Cell Proliferation
  • Cell Survival / drug effects*
  • Cyclin-Dependent Kinases / antagonists & inhibitors
  • Cyclin-Dependent Kinases / metabolism
  • Drug Evaluation, Preclinical
  • Female
  • Gene Expression Regulation, Leukemic / drug effects
  • Humans
  • Inhibitory Concentration 50
  • Interleukin-4 / physiology
  • Leukemia, Lymphocytic, Chronic, B-Cell / drug therapy*
  • Male
  • Middle Aged
  • Myeloid Cell Leukemia Sequence 1 Protein
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / metabolism*
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Purines / pharmacology*
  • Pyridines / pharmacology*
  • RNA Polymerase II / antagonists & inhibitors
  • Roscovitine
  • Signal Transduction / drug effects
  • Tumor Cells, Cultured / drug effects
  • X-Linked Inhibitor of Apoptosis Protein / genetics
  • X-Linked Inhibitor of Apoptosis Protein / metabolism

Substances

  • Antineoplastic Agents
  • CR8 compound
  • IL4 protein, human
  • Myeloid Cell Leukemia Sequence 1 Protein
  • NF-kappa B
  • Proto-Oncogene Proteins c-bcl-2
  • Purines
  • Pyridines
  • X-Linked Inhibitor of Apoptosis Protein
  • XIAP protein, human
  • Roscovitine
  • CD40 Ligand
  • Interleukin-4
  • Cyclin-Dependent Kinases
  • RNA Polymerase II