The CDK inhibitor purvalanol A induces neutrophil apoptosis and increases the turnover rate of Mcl‐1: potential role of p38‐MAPK in regulation of Mcl‐1 turnover

• Published in: Clinical and experimental immunology Vol. 192; no. 2; pp. 171 - 180
• Main Authors: Phoomvuthisarn, P., Cross, A., Glennon‐Alty, L., Wright, H. L., Edwards, S. W.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01-05-2018; John Wiley and Sons Inc
• Subjects: Activation; AKT protein; Apoptosis; Apoptosis - drug effects; cell activation; Cell cycle; Cells, Cultured; Cyclin-dependent kinase 2; DNA biosynthesis; Extracellular signal-regulated kinase; Gene Expression Regulation; Granulocyte-Macrophage Colony-Stimulating Factor - metabolism; Healthy Volunteers; Humans; Kinases; Leukemia; Leukocytes (granulocytic); Leukocytes (mononuclear); Leukocytes (neutrophilic); Macrophages; MAP kinase; Monocytes - drug effects; Myeloid Cell Leukemia Sequence 1 Protein - genetics; Myeloid Cell Leukemia Sequence 1 Protein - metabolism; Naphthalenes - pharmacology; Neutrophils; Neutrophils - drug effects; Neutrophils - pathology; Original; p38 Mitogen-Activated Protein Kinases - genetics; p38 Mitogen-Activated Protein Kinases - metabolism; Peripheral blood mononuclear cells; protein kinases/phophatases; Proteins; Proto-Oncogene Proteins c-akt - metabolism; Purines - pharmacology; Pyrazoles - pharmacology; Signal Transduction - drug effects; Transcription; Turnover rate; apoptosis; protein kinases/phophatases; neutrophils; cell activation
• ISSN: 0009-9104; 1365-2249
• DOI: 10.1111/cei.13107

Summary
Human neutrophils are terminally differentiated cells that do not replicate and yet express a number of enzymes, notably cell cycle‐dependent kinases (CDKs), that are associated normally with control of DNA synthesis and cell cycle progression. In neutrophils, CDKs appear to function mainly to regulate apoptosis, although the mechanisms by which they regulate this process are largely unknown. Here we show that the CDK2 inhibitor, purvalanol A, induces a rapid decrease in myeloid cell leukaemia factor‐1 (Mcl‐1) levels in human neutrophils and peripheral blood mononuclear cells (PBMCs), but only induces apoptosis in neutrophils which are dependent upon expression on this protein for survival. This rapid decrease in cellular Mcl‐1 protein levels was due to a purvalanol A‐induced decrease in stability, with the half‐life of the protein decreasing from approximately 2 h in control cells to just over 1 h after addition of the CDK2 inhibitor: it also blocked the granulocyte–macrophage colony‐stimulating factor (GM‐CSF)‐dependent stabilization of Mcl‐1. Purvanalol A blocked GM‐CSF‐stimulated activation of extracellular‐regulated kinase (Erk) and signal transducer and activator of transcription (STAT)‐3, and stimulated an additive activation of protein kinase B (Akt) with GM‐CSF. Purvalanol A alone stimulated a rapid and sustained activation of p38‐mitogen‐activated protein kinase (MAPK) and the pan p38‐MAPK inhibitor, BIRB796, partly blocked the purvalanol A‐induced apoptosis and Mcl‐1 loss. These novel effects of purvalanol A may result, at least in part, from blocking GM‐CSF‐mediated Erk activation. In addition, we propose that purvalanol A‐induced activation of p38‐MAPK is, at least in part, responsible for its rapid effects on Mcl‐1 turnover and acceleration of neutrophil apoptosis.
Human neutrophils do not replicate yet they express a number of proteins and enzymes that are normally required for cell cycle progression. Here we show that the CDK 2 inhibitor, Purvalanol A, induces a rapid induction of apoptosis of neutrophils (but not PBMCs) that is mediated via increased turnover of the anti‐apoptotic protein, Mcl‐1.