Assaying pharmacodynamic endpoints with targeted therapy: Flavopiridol and 17AAG induced dephosphorylation of histone H1.5 in acute myeloid leukemia

作者：Wang Liwen; Harshman Sean W; Liu Shujun; Ren Chen; Xu Hua; Sallans Larry; Grever Michael; Byrd John C; Marcucci Guido; Freitas Michael A^*

来源：Proteomics, 2010, 10(23): 4281-4292.

DOI：10.1002/pmic.201000080

• Acute myeloid leukemia
• Biomedicine
• Chemotherapy
• Histone
• Phosphorylation

Histone H1 is commonly used to assay kinase activity in vitro As many promising targeted therapies affect kinase activity of specific enzymes involved in cancer transformation H1 phosphorylation can serve as potential pharmacodynamic marker for drug activity within the cell In this study we utilized a phosphoproteomic workflow to characterize histone H1 phosphorylation changes associated with two targeted therapies in the Kasumi 1 acute myeloid leukemia cell line The phosphoproteomic workflow was first validated with standard casein phospho proteins and then applied to the direct analysis of histone HI from Kasumi 1 nuclear lysates Ten H1 phosphorylation sites were identified on the H1 variants H1 2 H1 3 Hi 4 H1 5 and H1 x LC MS profiling of intact His demonstrated global dephosphorylation of H1 5 associated with therapy by the cyclin dependent kinase inhibitor flavopiridol and the Heat Shock Protein 90 inhibitor 17 (Allylamino) 17 demethoxygeldanamycin In contrast independent treatments with a nucleotide analog proteosome inhibitor and histone deacetylase inhibitor did not exhibit decreased H1 5 phosphorylation The data presented herein demonstrate that potential of histones to assess tin cellular response of reagents that have direct and indirect effects on kinase activity that alters by tone phosphorylation As such this approach may be a highly informative marker for response to targeted therapies influencing histone phosphorylation

• 出版日期2010-12