RNAi screen of the druggable genome identifies modulators of proteasome inhibitor sensitivity in myeloma including CDK5

Blood. 2011 Apr 7;117(14):3847-57. doi: 10.1182/blood-2010-08-304022. Epub 2011 Feb 2.

Abstract

The molecular target(s) cooperating with proteasome inhibition in multiple myeloma (MM) remain unknown. We therefore measured proliferation in MM cells transfected with 13 984 small interfering RNAs in the absence or presence of increasing concentrations of bortezomib. We identified 37 genes, which when silenced, are not directly cytotoxic but do synergistically potentiate the growth inhibitory effects of bortezomib. To focus on bortezomib sensitizers, genes that also sensitized MM to melphalan were excluded. When suppressed, the strongest bortezomib sensitizers were the proteasome subunits PSMA5, PSMB2, PSMB3, and PSMB7 providing internal validation, but others included BAZ1B, CDK5, CDC42SE2, MDM4, NME7, RAB8B, TFE3, TNFAIP3, TNK1, TOP1, VAMP2, and YY1. The strongest hit CDK5 also featured prominently in pathway analysis of primary screen data. Cyclin-dependent kinase 5 (CDK5) is expressed at high levels in MM and neural tissues with relatively low expression in other organs. Viral shRNA knockdown of CDK5 consistently sensitized 5 genetically variable MM cell lines to proteasome inhibitors (bortezomib and carfilzomib). Small-molecule CDK5 inhibitors were demonstrated to synergize with bortezomib to induce cytotoxicity of primary myeloma cells and myeloma cell lines. CDK5 regulation of proteasome subunit PSMB5 was identified as a probable route to sensitization.

Publication types

  • Validation Study

MeSH terms

  • Antineoplastic Agents / administration & dosage
  • Antineoplastic Agents / isolation & purification
  • Antineoplastic Agents / pharmacology*
  • Boronic Acids / administration & dosage
  • Boronic Acids / pharmacology
  • Bortezomib
  • Cyclin-Dependent Kinase 5 / antagonists & inhibitors
  • Cyclin-Dependent Kinase 5 / genetics
  • Cyclin-Dependent Kinase 5 / isolation & purification
  • Cyclin-Dependent Kinase 5 / physiology*
  • Drug Resistance, Neoplasm / drug effects
  • Drug Resistance, Neoplasm / genetics*
  • Drug Screening Assays, Antitumor / methods
  • Drug Synergism
  • Enzyme Inhibitors / administration & dosage
  • Enzyme Inhibitors / pharmacology
  • Enzyme Inhibitors / therapeutic use*
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic / drug effects
  • Gene Regulatory Networks
  • Genome, Human / drug effects
  • High-Throughput Screening Assays
  • Humans
  • Microarray Analysis
  • Multiple Myeloma / drug therapy
  • Multiple Myeloma / genetics*
  • Multiple Myeloma / metabolism
  • Proteasome Endopeptidase Complex / genetics
  • Proteasome Endopeptidase Complex / metabolism
  • Proteasome Endopeptidase Complex / physiology
  • Proteasome Inhibitors*
  • Pyrazines / administration & dosage
  • Pyrazines / pharmacology
  • RNA Interference / physiology
  • RNA, Small Interfering / analysis
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / isolation & purification*
  • Tumor Cells, Cultured

Substances

  • Antineoplastic Agents
  • Boronic Acids
  • Enzyme Inhibitors
  • Proteasome Inhibitors
  • Pyrazines
  • RNA, Small Interfering
  • Bortezomib
  • Cyclin-Dependent Kinase 5
  • CDK5 protein, human
  • PSMB5 protein, human
  • Proteasome Endopeptidase Complex