Multiplexed phosphospecific flow cytometry enables large-scale signaling profiling and drug screening in blood platelets

Journal of Thrombosis and Haemostasis

Volumn 12, Issue 10, 2014, Pages 1733-1743

  Spurgeon, B E J ^a   Aburima, A ^a   Oberprieler, N G ^b   Tasken K ^b,c   Naseem, K M ^a  

^a UNIVERSITY OF HULL   (United Kingdom)

^b OSLO UNIVERSITY HOSPITAL   (Norway)

^c UNIVERSITY OF OSLO   (Norway)

Author keywords

Flow cytometry; Phosphorylation; Platelets; Protein kinase A; Signal transduction

Indexed keywords

3 BENZYL 5 (6 CARBOXYHEXYL) 1 (2 CYCLOHEXYL 2 HYDROXYETHYLAMINO)HYDANTOIN; ANTITHROMBOCYTIC AGENT; CELL PROTEIN; CYCLIC AMP; PHOSPHOSPECIFIC ANTIBODY; PROSTAGLANDIN DERIVATIVE; PROSTAGLANDIN E1 DERIVATIVE; VASODILATOR STIMULATED PHOSPHOPROTEIN; ACTIN BINDING PROTEIN; ANTIBODY; CELL ADHESION MOLECULE; CYCLIC AMP DEPENDENT PROTEIN KINASE; FLUORESCENT DYE; PHOSPHOPROTEIN; PROSTAGLANDIN; VASODILATOR-STIMULATED PHOSPHOPROTEIN;

ARTICLE; CONTROLLED STUDY; FLOW CYTOMETER; FLOW CYTOMETRY; FOLLOW UP; HIGH THROUGHPUT SCREENING; HUMAN; HUMAN CELL; IC50; PRIORITY JOURNAL; PROTEIN PHOSPHORYLATION; QUANTITATIVE ANALYSIS; SIGNAL TRANSDUCTION; THROMBOCYTE AGGREGATION; THROMBOCYTE AGGREGATION INHIBITION; THROMBOCYTE FUNCTION; ANIMAL; CHEMISTRY; DRUG DESIGN; DRUG EFFECTS; DRUG SCREENING; ELECTROPHORESIS; METABOLISM; MOUSE; PHOSPHORYLATION; THROMBOCYTE;

ANIMALS; ANTIBODIES; BLOOD PLATELETS; CELL ADHESION MOLECULES; CYCLIC AMP; CYCLIC AMP-DEPENDENT PROTEIN KINASES; DRUG DESIGN; DRUG EVALUATION, PRECLINICAL; ELECTROPHORESIS; FLOW CYTOMETRY; FLUORESCENT DYES; HUMANS; MICE; MICROFILAMENT PROTEINS; PHOSPHOPROTEINS; PHOSPHORYLATION; PLATELET AGGREGATION; PLATELET AGGREGATION INHIBITORS; PROSTAGLANDINS; SIGNAL TRANSDUCTION;

EID: 84907904401     PISSN: 15387933     EISSN: 15387836     Source Type: Journal    
DOI: 10.1111/jth.12670     Document Type: Article

References (23)

1. Jackson SP. Arterial thrombosis - insidious, unpredictable and deadly. Nat Med 2011; 17: 1423-36.
2. Dittrich M, Birschmann I, Mietner S, Sickmann A, Walter U, Dandekar T. Platelet protein interactions: map, signaling components, and phosphorylation groundstate. Arterioscler Thromb Vasc Biol 2008; 28: 1326-31.
3. Krutzik PO, Nolan GP. Fluorescent cell barcoding in flow cytometry allows high-throughput drug screening and signaling profiling. J Immunol 2006; 3: 1581-9.
4. Krutzik PO, Crane JM, Clutter MR, Nolan GP. High-content single-cell drug screening with phosphospecific flow cytometry. Nat Chem Biol 2008; 4: 132-42.
5. Butt E, Abel K, Krieger M, Palm D, Hoppe V, Hoppe J, Walter U. cAMP- and cGMP-dependent protein kinase phosphorylation sites of the focal adhesion vasodilator-stimulated phosphoprotein (VASP) in vitro and in intact human platelets. J Biol Chem 1994; 269: 14509-17.
6. Michelson AD. Advances in antiplatelet therapy. Hematology Am Soc Hematol Educ Program. 2011; 2011: 62-9.
7. Gachet C, Aleil B. Testing antiplatelet therapy. Eur Hear J Suppl 2008; 10: A28-34.
8. Harrison P, Mumford A. Screening tests of platelet function: update on their appropriate uses for diagnostic testing. Semin Thromb Hemost 2009; 35: 150-7.
9. Glenn JR, Dovlatova N, White AE, Dhillon K, Heptinstall S, Fox SC. 'VASPFix' for measurement of VASP phosphorylation in platelets and for monitoring effects of P2Y12 antagonists. Thromb Haemost 2014; 111: 539-48.
10. Roberts W, Magwenzi S, Aburima A, Naseem KM, Dc W. Thrombospondin-1 induces platelet activation through CD36-dependent inhibition of the cAMP/protein kinase A signaling cascade. Blood 2010; 116: 4297-306.
11. Aburima A, Wraith KS, Raslan Z, Law R, Magwenzi S, Naseem KM. cAMP signaling regulates platelet myosin light chain (MLC) phosphorylation and shape change through targeting the RhoA-Rho kinase-MLC phosphatase signalling pathway. Blood 2013; 122: 3533-45.
12. Kotecha N, Krutzik PO, Irish JM. Web-based analysis and publication of flow cytometry experiments. Curr Prot Cytom 2010; 53: 10.17.1-10.17.24.
13. Schwarz UR, Geiger J, Walter U, Eigenthaler M. Flow cytometry analysis of intracellular VASP phosphorylation for the assessment of activating and inhibitory signal transduction pathways in human platelets - definition and detection of ticlopidine/clopidogrel effects. Thromb Haemost 1999; 82: 1145-52.
14. Oberprieler NG, Lemeer S, Kalland ME, Torgersen KM, Heck AJR, Taskén K. High-resolution mapping of prostaglandinE2-dependent signaling networks identifies a constitutively active PKA signaling node in CD8+CD45RO+ T cells. Blood 2010; 116: 2253-65.
15. Seamon KB, Daly JW. Forskolin: a unique diterpene activator of cyclic AMP-generating systems. J Cyclic Nucleotide Res 1981; 7: 201-24.
16. Fredholm BB. On the mechanism of action of theophylline and caffeine. Acta Med Scand 1985; 217: 149-53.
17. Podzuweit T, Nennstiel P, Müller A. Isozyme selective inhibition of cGMP-stimulated cyclic nucleotide phosphodiesterases by erythro-9-(2-hydroxy-3-nonyl) adenine. Cell Signal 1995; 7: 733-8.
18. Tang KM, Jang EK, Haslam RJ. Photoaffinity labelling of cyclic GMP-inhibited phosphodiesterase (PDE III) in human and rat platelets and rat tissues: effects of phosphodiesterase inhibitors. Eur J Pharmacol 1994; 268: 105-14.
19. Corbin JD, Francis SH. Pharmacology of phosphodiesterase-5 inhibitors. Int J Clin Pract 2002; 56: 453-9.
20. Haslam RJ, Dickinson NT, Jang EK. Cyclic nucleotides and phosphodiesterases in platelets. Thromb Haemost 1999; 82: 412-23.
21. Zhang J, Chung T, Oldenburg K. A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 1999; 4: 67-73.
22. Irish JM, Czerwinski DK, Nolan GP, Levy R. Kinetics of B cell receptor signaling in human B cell subsets mapped by phosphospecific flow cytometry. J Immunol 2006; 177: 1581-9.
23. Irish JM, Hovland R, Krutzik PO, Perez OD, Bruserud Ø, Gjertsen BT, Nolan GP. Single cell profiling of potentiated phospho-protein networks in cancer cells. Cell 2004; 118: 217-28.