Label-free cell phenotypic drug discovery

Combinatorial Chemistry and High Throughput Screening

Volumn 17, Issue 7, 2014, Pages 566-578

  Fang, Ye ^a  

^a Biochemical Technologies Science and Technology Division Corning Incorporated ^*  (United States)

Author keywords

Label free cell phenotypic screen; Phenotypic screen; Polypharmacology; Target directed drug discovery

Indexed keywords

ARTICLE; BACTERIAL CELL; BIOSENSOR; CANCER STEM CELL; CELL ADHESION; CELL CYCLE; CELL DIFFERENTIATION; CELL FUNCTION; CELL LABELING; DRUG DEVELOPMENT; DRUG TARGETING; ENDOTHELIUM CELL; EXTRACELLULAR MATRIX; HIGH THROUGHPUT SCREENING; HUMAN; NONHUMAN; PHENOTYPE; POLYPHARMACOLOGY; PRIORITY JOURNAL; PRODUCTIVITY; SIGNAL TRANSDUCTION; ANIMAL; DEVICES; EQUIPMENT DESIGN; GENETIC PROCEDURES; PROCEDURES;

ANIMALS; BIOSENSING TECHNIQUES; DRUG DISCOVERY; EQUIPMENT DESIGN; HUMANS; PHENOTYPE; POLYPHARMACOLOGY;

EID: 84906268621     PISSN: 13862073     EISSN: 18755402     Source Type: Journal    
DOI: 10.2174/1386207317666140211100000     Document Type: Article

References (134)

1. Hart, C.P. Finding the target after screening the phenotype. Drug Discov. Today, 2005, 10, 513-519
2. Kenakin, T. Cellular assays as portals to seven-transmembrane receptor-based drug discovery. Nat. Rev. Drug Discov., 2009, 8, 617-626
3. Bleicher, K.H.; Böhm, H.J.; Müller, K.; Alanine, A.I. Hit and lead generation: beyond high-throughput screening. Nat. Rev. Drug Discov., 2003, 2, 369-378
4. Macarron, R.; Banks, M.N.; Bojanic, D.; Burns, D.J.; Cirovic, D.A.; Garyantes, T.; Green, D.V.; Hertzberg, R.P.; Janzen, W.P.; Paslay, J.W.; Schopfer, U.; Sittampalam, G.S. Impact of highthroughput screening in biomedical research. Nat. Rev. Drug Discov., 2011, 10, 188-195
5. Booth, B.; Zemmel, R. Prospects for productivity. Nat. Rev. Drug Discov., 2004, 3, 451-456
6. Paul, S.M.; Mytelka, D.S.; Dunwiddie, C.T.; Persinger, C.C.; Munos, B.H.; Lindborg, S.R.; Schacht, A.L. How to improve R&D productivity: the pharmaceutical industry's grand challenge. Nat. Rev. Drug Discov., 2010, 9, 203-214
7. Pammolli, F.; Magazzini, L.; Riccaboni, M. The productivity crisis in pharmaceutical R&D. Nat. Rev. Drug Discov., 2011, 10, 428-438
8. Scannell, J.W. Diagnosing the decline in pharamaceutical R&D efficiency. Nat. Rev. Drug Discov., 2012, 11, 191-200.
9. Clemons, P.A. Complex phenotypic assays in high-throughput screening. Curr. Opin. Chem. Biol., 2004, 8, 334-338
10. Swinney, D.C.; Anthony, J. How were new medicines discovered? Nat. Rev. Drug Discov., 2011, 10, 507-519
11. Lee, J.A.; Uhlik, M.T.; Moxham, C.M.; Tomandl, D.; Sall, D.J. Modern phenotypic drug discovery is a viable, neoclassic pharma strategy. J. Med. Chem., 2012, 55, 4527-4538
12. Swinney, D.C. Phenotypic vs target-based drug discovery for firstin- class medicines. Clin. Pharmacol. Ther., 2013, 93, 299-301
13. Eggert, U.S. The why and how of phenotypic small-molecule screens. Nat. Chem. Biol., 2013, 9, 206-209
14. Hopkins, A.L. Network pharmacology: the next paradigm in drug discovery. Nat. Chem. Biol., 2008, 4, 682-690
15. Knight, Z.A.; Lin, H.; Shokat, K.M. Targeting the cancer kinome through polypharmacology. Nat. Rev. Cancer, 2010, 10, 130-137
16. Medina-Franco, J.L.; Giulianotti, M.A.; Welmaker, G.S.; Houghten, R.A. Shifting from the single to the multitarget paradigm in drug discovery. Drug Discov. Today, 2013, 18, 495-501
17. Rask-Andersen, M.; Almén, M.S.; Schiöth, H.B. Trends in the exploitation of novel drug targets. Nat. Rev. Drug Discov., 2011, 10, 579-590
18. Terstappen, G.C.; Schlüpen, C.; Raggiaschi, R.; Gaviraghi, G. Target deconvolution strategies in drug discovery. Nat. Rev. Drug Discov., 2007, 6, 891-903
19. Titov, D.V.; Liu, J.O. Identification and validation of protein targets of bioactive small molecules. Bioorg. Med. Chem., 2012, 20, 1902-1909
20. Lee, J.; Bogyo, M. Target deconvolution techniques in modern phenotypic profiling. Curr. Opin. Chem. Biol., 2013, 17, 118-126
21. Simon, G.M.; Niphakis, M.J.; Cravatt, B.F. Determining target engagement in living systems. Nat. Chem. Biol., 2013, 9, 200-205
22. Schenone, M.; Dancík, V.; Wagner, B.K.; Clemons, P.A. Target identification and mechanism of action in chemical biology and drug discovery. Nat. Chem. Biol., 2013, 9, 232-240
23. Fu, H.; Fu, W.; Sun, M.; Shou, Q.; Zhai, Y.; Cheng, H.; Teng, L.; Mou, X.; Li, Y.; Wan, S.; Zhang, S.; Xu, Q.; Zhang, X.; Wang, J.; Zhu, J.; Wang, X.; Xu, X.; Lv, G.; Jin, L.; Guo, W.; Ke, Y. Kinetic cellular phenotypic profiling: Prediction, identification, and analysis of bioactive natural products. Anal. Chem., 2011, 83, 6518-6526
24. Fang, Y. The development of label-free cellular assays for drug discovery. Expert Opin. Drug Discov., 2011, 6, 1285-1298
25. Fang, Y. Troubleshooting and deconvoluting label-free cell phenotypic assays in drug discovery. J. Pharmacol. Tox. Methods, 2013, 67, 69-81
26. Fang, Y. Label-free biosensors for cell biology. International Journal of Electrochemistry, 2011, 2011, e460850
27. Ferrie, A.M.; Wang, C.;, Deng, H.;, Fang, Y. Label-free optical biosensor with microfluidics identifies an intracellular signalling wave mediated through the ß 2-Adrerengic receptor. Integr. Biol., 2013, 5, 1253-1261
28. Munos, B. Lessons from 60 years of pharmaceutical innovation. Nat. Rev. Drug Discov., 2009, 8, 959-968
29. David, E.; Tramontin, T.; Zemmel, R. Pharmaceutical R&D: the road to positive returns. Nat. Rev. Drug Discov., 2009, 8, 609-610
30. Zambrowicz, B.P.;, Sands, A.T. Knockouts model the 100 bestselling drugs-will they model the next 100? Nat. Rev. Drug Discov., 2003, 2, 38-51
31. Drews J. Drug discovery: a historical perspective. Science, 2000, 287,1960-1964
32. Imming, P.; Sinning, C.; Meyer A. Drugs, their targets and the nature and number of drug targets. Nat. Rev. Drug Discov., 2006, 5, 821-834
33. Overington, J. P.; Al-Lazikani, B.; Hopkins, A.L. How many drug targets are there? Nat. Rev. Drug Discov., 2006, 5, 993-996
34. Hopkins, A.L.; Groom, C.R. The druggable genome. Nat. Rev. Drug Discov., 2002, 1, 727-730
35. Yildirim, M.A.; Goh, K.I.; Cusick, M.E.; Barabasi, A.L.; Vidal, M. Drug-target network. Nat. Biotechnol., 2007, 25, 1119-1126
36. Roth, B.L.; Sheffler, D.J.; Kroeze, W.K. Magic shotguns versus magic bullets: selectively non-selective drugs for mood disorders and schizophrenia. Nat. Rev. Drug Discov., 2004, 3, 353-359
37. Strebhardt, K.; Ullrich, A. Paul Ehrlich's magic bullet concept: 100 years of progress. Nat. Rev. Cancer, 2008, 8, 473-480
38. Bell, D.W. Our changing view of the genomic landscape of cancer. J. Pathology, 2010, 220, 231-243
39. Fang, Y. Probing cancer signaling with resonant waveguide grating biosensors. Expert Opin. Drug Discov., 2010, 5, 1237-1248
40. Hillenmeyer, M.E., Fung, E.; Wildenhain, J.; Pierce, S.E.; Hoon, S.; Lee, W.; Proctor, M.; St Onge R.P.; Tyers, M.; Koller, D.; Altman, R.B.;, Davis, R.W.; Nislow, C.; Giaever, G. The chemical genomic portrait of yeast: uncovering a phenotype for all genes. Science, 2008, 320, 362-365
41. Frantz, S. Drug discovery: playing dirty. Nature, 2005, 437, 942-943
42. Kamb, A.; Wee, S.; Lengauer, C. Why is cancer drug discovery so difficult? Nat. Rev. Drug Discov., 2007, 6, 115-120
43. Keith, C.T.; Borisy, A.A.; Stockwell, B.R. Multicomponent therapeutics for networked systems. Nat. Rev. Drug Discov., 2005, 4, 1-8
44. Schadt, E.C.; Friend, S.H.; Shaywitz, D.A. A network view of disease and compound screening. Nat. Rev. Drug Discov., 2009, 8, 286-295
45. Collins, J.; Workman, P. New approaches to molecular cancer therapeutics. Nat. Chem. Biol., 2006, 2, 689-700
46. Anastassiadis, T.; Deacon, S.W.; Devarajan, K.; Ma, H.; Peterson, J.R. Comprehensive assay of kinase catalytic activity reveals feature of kinase inhibitor selectivity. Nat. Biotechnol., 2011, 29, 1039-1045
47. Davis, M.I.; Hunt, J.P.; Herrgard, S.; Ciceri, P.; Wodicka, L.M.; Pallares, G.; Hocker, M.; Treiber, D.K.; Zarrinkar, P.P. Comprehensive analysis of kinase inhibitor selectivity. Nat. Biotechnol., 2011, 29, 1046-1051
48. Wilhelm, S.M.; Adnane, L.; Newell, P.; Villanueva, A.; Llovet, J.M.; Lynch, M. Preclinical overview of sorafenib, a multikinase inhibitor that targets both Raf and VEGF and PDGF receptor tyrosine kinase signaling. Mol. Cancer Ther., 2008, 7, 3129-3140
49. Lin, X.; Huang, X.P.; Chen, G.; Whaley, R.; Peng, S.; Wang, Y.; Zhang, G.; Wang, S.X.; Wang, S.; Roth, B.L.; Huang, N. Life beyond kinases: structure-based discovery of sorafenib as nanomolar antagonist of 5-HT receptors. J. Med. Chem., 2012, 55, 5749-5759
50. Hargrave-Thomas, E.; Yu, B.; Reynisson, J. Serendipity in anticancer drug discovery. World J. Clin. Oncol., 2012, 3, 1-6
51. Vézina, C.; Kudelski, A.; Sehgal, S.N. Rapamycin (AY-22,989), a new antifungal antibiotic. J. Antibiot., 1975, 28,721-726
52. Morelon, E.; Stern, M.; Israël-Biet, D.; Correas, J.M.; Danel, C.; Mamzer-Bruneel, M.F.; Peraldi, M.N.; Kreis, H. Characteristics of sirolimus-Associated interstitial pneumonitis in renal transplant patients. Transplantation, 2001, 72, 787-790
53. Knox, C.; Law, V.; Jewison, T.; Liu, P.; Ly, S.; Frolkis, A.; Pon, A.; Banco, K.; Mak, C.; Neveu, V.; Djoumbou, Y.; Eisner, R.; Guo, A.C.; Wishart, D.S. DrugBank 3.0: a comprehensive resource for 'omics' research on drugs. Nucleic Acids Res., 2011, 39(Database issue), D1035-1041
54. Cooper, C.R. Development of Mycobacterium tuberculosis whole cell screening hits as potential antituberculosis agents. J. Med. Chem., 2013, 56, 7755-7760
55. Roemer, T.; Boone, C. Systems-level antimicrobial drug and drug synergy discovery. Nat. Chem. Biol., 2013, 9, 222-231
56. Nichols, R.J.; Sen, S.; Choo, Y.J.; Beltrao, P.; Zietek, M.;, Chaba, R.; Lee, S.;, Kazmierczak, K.M.; Lee, K.J.; Wong, A.; Shales, M.; Lovett, S.; Winkler, M.E.; Krogan, N.J.; Typas, A.;, Gross, C.A. Phenotypic landscape of a bacterial cell. Cell, 2011, 144, 143-156
57. Roemer, T.; Davies, J.; Giaever, G.; Nislow, C. Bugs, drugs and chemical genomics. Nat. Chem. Biol., 2011, 8, 46-56
58. Guiguemde, W.A.; Shelat, A.A.; Garcia-Bustos, J.F.; Diagana, T.T.; Gamo, F.J.; Guy, R.K. Global phenotypic screening for antimalarials. Chem. Biol., 2012, 19, 116-129
59. Chong, C.R.; Chen, X.; Shi, L.; Liu, J.O.;, Sullivan, D.J. Jr. A clinical drug library screen identifies astemizole as an antimalarial agent. Nat. Chem. Biol., 2006, 2, 415-416
60. Welsh, M.; Mangravite, L.; Medina, M.W.; Tantisira, K.; Zhang, W.; Huang, R.S.; McLeod, H.; Dolan, M.E. Pharmacogenomic discovery using cell-based models. Pharmacol. Rev., 2009, 61, 413-429
61. Kepp, O.; Galluzzi, L.;, Lipinski, M.; Yuan, J.; Kroemer, G. Cell death assays for drug discovery. Nat. Rev. Drug Discov., 2011, 10, 221-237
62. Kwon, H. J.; Kim, M. S.; Kim, M. J.; Nakajima, H.; Kim, K.-W. Histone deacetylase inhibitor FK228 inhibits tumor angiogenesis. Int. J. Cancer, 2002, 97, 290-296
63. Lee, J.A.; Uhlik, M.T.; Moxham, C.M.; Tomandl, D.; Sall, D.J. Modern phenotypic drug discovery is a viable, neoclassic pharma strategy. J. Med. Chem., 2012, 55, 4527-4538
64. Feng, Y.; Mitchison, T.J.; Bender, A.; Young, D.W.; Tallarico, J.A. Multi-parameter phenotypic profiling: using cellular effects to characterize small-molecule compounds. Nat. Rev. Drug Discov., 2009, 8, 567-578
65. Young, D.W.; Bender, A.; Hoyt, J.; McWhinnie, E.; Chirn, G.W.; Tao, C.Y.; Tallarico, J.A.; Labow, M.; Jenkins, J.L.; Mitchison, T.J.; Feng, Y. Integrating high-content screening and ligand-target prediction to identify mechanism of action. Nat. Chem. Biol., 2008, 4, 59-68
66. Walter, T.; Shattuck, D.W.; Baldock, R.; Bastin, M.E.; Carpenter, A.E.; Duce, S.; Ellenberg, J.; Fraser, A.; Hamilton, N.; Pieper, S.; Ragan, M.A.; Schneider, J.E.; Tomancak, P.; Hériché, J.K. Visualization of image data from cells to organisms. Nat. Methods., 2010, 7(3 Suppl): S26-41
67. Hartwell, K.A.; Miller, P.G.; Mukherjee, S.; Kahn, A.R.; Stewart, A.L.; Logan, D.J.; Negri, J.M.; Duvet, M.; Järås, M.; Puram, R.; Dancik, V.; Al-Shahrour, F.; Kindler, T.; Tothova, Z.; Chattopadhyay, S.; Hasaka, T.; Narayan, R.; Dai, M.; Huang, C.; Shterental, S.; Chu, L.P.; Haydu, J.E.; Shieh, J.H.; Steensma, D.P.; Munoz, B.; Bittker, J.A.; Shamji, A.F.; Clemons, P.A.; Tolliday, N.J.; Carpenter, A.E.; Gilliland, D.G.; Stern, A.M.; Moore, M.A.; Scadden, D.T.; Schreiber, S.L.; Ebert, B.L.; Golub, T.R. Nichebased screening identifies small-molecule inhibitors of leukemia stem cells. Nat. Chem. Biol., 2013, 9, 840-848
68. Sachlos, E.; Risueño, R.M.; Laronde, S.; Shapovalova, Z.; Lee, J.H.; Russell, J.; Malig, M.; McNicol, J.D.; Fiebig-Comyn, A.; Graham, M.; Levadoux-Martin, M.; Lee, J.B.; Giacomelli, A.O.; Hassell, J.A.; Fischer-Russell, D.; Trus, M.R.; Foley, R.; Leber, B.; Xenocostas, A.; Brown, E.D.; Collins, T.J.; Bhatia, M. Identification of drugs including a dopamine receptor antagonist that selectively target cancer stem cells. Cell, 2012, 149, 1284-1297
69. Gasque, G.; Conway, S.; Huang, J.; Rao, Y.; Vosshall, L.B. Small molecule drug screening in Drosophila identifies the 5HT2A receptor as a feeding modulation target. Sci. Rep., 2013, 3, 2120
70. Dar, A.C.; Das, T.K.; Shokat, K.M.; Cagan, R.L. Chemical genetic discovery of targets and anti-targets for cancer polypharmacology. Nature, 2012, 486, 80-84
71. MacRae, C.A.; Peterson, R.T. Zebrafish-based small molecule discovery. Chem. Biol., 2003, 10, 901-908
72. Zon, L.I.; Peterson, R.T. In vivo drug discovery in the zebrafish. Nat. Rev. Drug Discov., 2005, 4, 35-44
73. Kokel, D.; Bryan, J.; Laggner, C.; White, R.; Cheung, C.Y.; Mateus, R.; Healey, D.; Kim, S.; Werdich, A.A.; Haggarty, S.J.; Macrae, C.A.; Shoichet, B.; Peterson, R.T. Rapid behavior-based identification of neuroactive small molecules in the zebrafish. Nat. Chem. Biol., 2010, 6, 231-237
74. Fang, Y. Ligand-receptor interaction platforms and their applications for drug discovery. Expert Opin. Drug Discov., 2012, 7, 969-988
75. Pal, S.; Fauchet, P.M.; Miller, B.L. 1-D and 2-D photonic crystals as optical methods for 'amplifying' biomolecular recognition. Anal. Chem., 2012, 84, 8900-8908
76. Fang, Y. Label-free cell-based assays with optical biosensors in drug discovery. Assay Drug Dev. Technol., 2006, 4, 583-595
77. Fang, Y.; Ferrie, A.M.; Fontaine, N.H.; Mauro, J.; Balakrishnan, J. Resonant waveguide grating biosensor for living cell sensing. Biophys. J., 2006, 91, 1925-1940
78. Wegener, J.; Keese, C.R.; Giaever, I. Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces. Exp. Cell Res., 2000, 259, 158-166
79. Fang, Y. Label-free receptor assays. Drug Discov. Today Technol., 2010, 7, e5-e11
80. Yu, N.; Atienza, J.M.; Bernard, J.; Blanc, S.; Zhu, J.; Wang, X.; Xu, X.; Abassi, Y.A. Real-time monitoring of morphological changes in living cells by electronic cell sensor arrays: An approach to study G protein-coupled receptors. Anal. Chem., 2006, 78, 35-43
81. Schröder, R.; Janssen, N.; Schmidt, J.; Kebig, A.; Merten, N.; Hennen, S.; Müller, A.; Blättermann, S.; Mohr-Andrä, M.; Zahn, S.; Wenzel, J.; Smith, N.J. Gomeza, J.; Drewke, C.; Milligan, G.; Mohr, K.; Kostenis, E. Deconvolution of complex G proteincoupled receptor signaling in live cells using dynamic mass redistribution measurements. Nat. Biotechnol., 2010, 28, 943-949
82. Hennen, S.; Wang, H.; Peters, L.; Merten, N; Simon, K.; Spinrath, A.; Blättermann, S.; Akkari, R.; Schrage, R.; Schröder, R.; Schulz, D.; Vermeiren, C.; Zimmermann, K.; Kehraus, S.;, Drewke, C.; Pfeifer, A.; König, G.M.; Mohr, K.; Gillard, M.; Müller, C.E.; Lu, Q.R.; Gomeza, J.; Kostenis, E. Decoding signaling and function of the orphan G protein-coupled receptor GPR17 with a smallmolecule agonist. Sci. Signal., 2013, 6, ra93
83. Bagnaninchi, P. O.; Drummond, N. Real-time label-free monitoring of adipose-derived stem cell differentiation with electric cellsubstrate impedance sensing. Proc. Natl. Acad. Sci. U.S.A., 2011, 108, 6462-6467
84. Abassi, Y.A.; Xi, B.; Li, N.; Ouyang, W.; Seiler, A.; Watzele, M.; Kettenhofen, R.; Bohlen, H.; Ehlich, A.; Kolossov, E.; Wang, X.; Xu, X. Dynamic monitoring of beating periodicity of stem cellderived cardiomyocytes as a predictive tool for preclinical safety assessment. Br. J. Pharmacol., 2012, 165, 1424-1441
85. Pai, S.; Verrier, F.; Sun, H.; Hu, H.; Ferrie, A. M.; Eshraghi, A.; Fang, Y. Dynamic mass redistribution assay decodes differentiation of a neural progenitor stem cell. J. Biomol. Screen., 2012, 17, 1180-1191
86. Ferrie, A.M.; Sun, H.; Zaytseva, N.; Fang, Y. Divergent label-free cell phenotypic pharmacology of ligands at the overexpressed ß2- adrenergic receptors. Sci. Rep., 2014, 4, 3828
87. Ferrie, A.M.; Sun, H.; Fang, Y. Label-free integrative pharmacology on-target of drugs at the ß2-Adrenergic receptor. Sci. Rep., 2011, 1, 33
88. Morse, M.; Tran, E.; Levension, R.L.; Fang, Y. Ligand-directed functional selectivity at the mu opioid receptor revealed by labelfree on-target pharmacology. PLoS One, 2011, 6, e25643
89. Morse, M.; Sun, H.; Tran, E.; Levenson, R.; Fang, Y. Label-free integrative pharmacology ontarget of opioid ligands at the opioid receptor family. BMC Pharmacol. Tox., 2013, 14, 17
90. Verdonk, E.; Johnson, K.; McGuinness, R.; Leung, G.; Chen, Y.- W.; Tang, H.R.; Michelotti, J.M.; Liu, V.F. Cellular dielectric spectroscopy: a label-free comprehensive platform for functional evaluation of endogenous receptors. Assays Drug Dev. Technol., 2006, 4, 609-19
91. Tran, E.; Fang, Y. Duplexed label-free G protein-coupled receptor assays for high throughput screening. J. Biomol. Screen., 2008, 13, 975-985
92. Dodgson, K.; Gedge, L.; Murray, D.C.; Coldwell, M. A 100K well screen for a muscarinic receptor using the Epic label-free system - a reflection on the benefits of the label-free approach to screening seven-transmembrane receptors. J. Recept. Signal Transduct., 2009, 29, 163-72
93. Nelson, C.P.; Challiss, R.A. "Phenotypic" pharmacology: the influence of cellular environment on G protein-coupled receptor antagonist and inverse agonist pharmacology. Biochem. Pharmacol., 2007, 73, 737-751
94. Kinzer-Ursem, T.L.; Linderman, J.J. Both ligand- and cell-specific parameters control ligand agonism in a kinetic model of G proteincoupled receptor signaling. PLoS Comput. Biol., 2007, 3, e6
95. Wermuth, C.G. Multitarget drugs: the end of the 'one-target-onedisease' philosophy? Drug Discov. Today, 2004, 9, 826-827
96. Deng, H.; Hu, H.; Fang, Y. Multiple tyrosine metabolites are GPR35 agonists. Sci. Rep., 2012, 2, 373
97. Kenakin, T.; Miller, L.J. Seven transmembrane receptors as shapeshifting proteins: the impact of allosteric modulation and functional selectivity on new drug discovery. Pharmacol. Rev., 2010, 62, 265-304
98. Kenakin, T. The potential for selective pharmacological therapies through biased receptor signaling. BMC Pharmacol. Tox., 2012, 13, 3
99. Guo, D.; Mulder-Krieger, T.; Ijzerman, A.P.; Heitman, L.H. Functional efficacy of adenosine A2A receptor agonists is positively correlated to their receptor residence time. Br. J. Pharmacol., 2012, 166, 1846-1959
100. Deng, H.; Wang, C.; Su, M.; Fang, Y. Probing biochemical mechanisms of action of muscarinic M3 receptor antagonists with label-free whole-cell assays. Anal. Chem., 2012, 84, 8232-8239
101. Deng, H.; Sun, H.; Fang, Y. Label-free cell phenotypic efficacy of agonists at endogenous muscarinic M3 receptor correlates with their residence time. J. Pharmacol. Tox. Methods, 2013, 68, 323-333
102. Bock, A.; Chirinda, B.; Krebs, F.; Messerer, R.; Bätz, J.; Muth, M.; Dallanoce, C.; Klingenthal, D.; Tränkle, C.; Hoffmann, C.; De Amici, M.; Holzgrabe, U.; Kostenis, E.; Mohr, K. Dynamic ligand binding dictates partial agonism at a G protein-coupled receptor. Nat. Chem. Biol., 2014, 10, 18-20
103. Deng, H.; Wang, C.; Fang, Y. Label-free cell phenotypic assessment of the molecular mechanism of action of epidermal growth factor receptor inhibitors. RSC Adv., 2013, 3, 10370-10378
104. Ramsden, J.J.; Horvath, R. Optical biosensors for cell adhesion. J. Recept. Signal Transduct., 2009, 29, 211-223
105. Fang, Y. Label-free biosensor cellular assays for cell adhesion. J. Adhesion Sci. Technol., 2010, 24, 1011-1021
106. Zaytseva, Z.; Lynn, J.G.; Wu, Q.; Mudaliar, D.J.; Kuang, P.Q.; Fang, Y. Resonant waveguide grating biosensor-enabled label-free and fluorescence detection of cell adhesion. Sens. Actuat. B: Chem., 2013, 188, 1064-1072
107. Abassi, Y.A.; Xi, B.; Zhang, W.; Ye, P.; Kirstein, S.L.; Gaylord, M.R.; Feinstein, S.C.; Wang, X.; Xu, X. Kinetic cell-based morphological screening: prediction of mechanism of compound action and off-target effects. Chem. Biol., 2009, 16, 712-723
108. Ke, N.; Xi, B.; Ye, P.; Xu, W.; Zheng, M.; Mao, L.; Wu, M-J.; Zhu, J.; Wu, J.; Zhang, W.; Zhang, J.;, Irelan, J.; Wang, X.;, Xu, X.; Abassi, Y.A. Screening and identification of small molecule compounds perturbing mitosis using time-dependent cellular response profiles. Anal. Chem., 2010, 82, 6495-6503
109. Zhang, J.D.; Koerner, C.; Bechtel, S.; Bender, C.; Keklikoglou, I.; Schmidt, C.; Irsigler, A.; Ernst, U.; Sahin, O.; Wiemann, S.; Tschulena, U. Time-resolved human kinome RNAi screen identifies a network regulating mitotic-events as early regulators of cell proliferation. PLoS One, 2011, 6, e22176
110. Kholodenko, B.N. Cell signaling dynamics in time and space. Nat. Rev. Mol. Cell Biol., 2006, 7, 165-176
111. Scott, J.D.; Pawson, T. Cell signaling in space and time: where proteins come together and when they're apart. Science, 2009, 326, 1220-1224
112. Fang, Y.; Ferrie, A.M.; Fontaine, N.H.; Yuen, P.K. Characteristics of dynamic mass redistribution of EGF receptor signaling in living cells measured with label free optical biosensors. Anal. Chem., 2005, 77, 5720-5725
113. Calebiro, D.; Nikolaev, V.O.; Persani, L.; Lohse, M.J. Signaling by internalized G-protein-coupled receptors. Trends Pharmacol. Sci., 2010, 31, 221-228
114. Babcock, J.J.; Li, M. Inside job: Ligand-receptor pharmacology beneath the plasma membrane. Acta Pharmacol. Sinica, 2013, 34, 859-869
115. Deng, H.; Hu, H.; He, M.; Hu, J.; Niu, W.; Ferrie, A.M.; Fang, Y. Discovery of 2-(4-methylfuran-2(5H)-ylidene)malononitrile and thieno[3,2-b]thiophene-2-carboxylic acid derivatives as G proteincoupled receptor-35 (GPR35) agonists. J. Med. Chem., 2011, 54, 7385-7396
116. Deng, H.; Hu, H.; Fang, Y. Tyrphostin analogs are GPR35 agonists. FEBS Lett., 2011, 585, 1957-1962
117. Hu, H.; Deng, H.; Fang, Y. Label-free phenotypic profiling identified D-luciferin as a GPR35 agonist. PLoS One, 2012, 7, e34934
118. Fang, Y.; Li, G.; Ferrie, A.M. Non-invasive optical biosensor for assaying endogenous G protein-coupled receptors in adherent cells. J. Pharmacol. Tox. Methods, 2007, 55, 314-322
119. Fang, Y.; Ferrie, A.M. Label-free optical biosensor for liganddirected functional selectivity acting on ß2-Adrenoceptor in living cells. FEBS Lett., 2008, 582, 558-564
120. Lindholm-Sethson, B.; Nyström, J.; Malmsten, M.; Ringstad, L.; Nelson, A.; Geladi, P. Electrochemical impedance spectroscopy in label-free biosensor applications: Multivariate data analysis for an objective interpretation. Anal. Bioanal. Chem., 2010, 398, 2341-2349
121. Stallaert, W.; Dorn, J.F.; van der Westhuizen, E.; Audet, M.; Bouvier, M. Impedance responses reveal ß 2-Adrenergic receptor signaling pluridimensionality and allow classification of ligands with distinct signaling profiles. PLoS One, 2012, 7, e29420
122. Goral, V.; Jin, Y.; Sun, H.; Ferrie, A.M.; Wu, Q.; Fang, Y. Agonist-directed desensitization of the ß2-Adrenergic receptor. PLoS One, 2011, 6, e19282
123. Goral, V.; Wu, Q.; Sun, H.; Fang, Y. Label-free optical biosensor with microfluidics for sensing ligand-directed functional selectivity on trafficking of thrombin receptor. FEBS Lett., 2011, 585, 1054-1060
124. Zaytseva, N.; Miller, W.; Goral, V.; Hepburn, J.; Fang, Y. Microfluidic resonant waveguide grating biosensor system for whole cell sensing. Appl. Phys. Lett., 2011, 96, 163703
125. Copeland, R.A.; Pompliano, D.L.; Meek, T.D. Drug-target residence time and its implications for lead optimization. Nat. Rev. Drug Discov., 2006, 5, 730-739
126. Ziegler, S.; Pries, V.; Hedberg, C.; Waldmann, H. Target identification for small bioactive molecules: finding the needle in the haystack. Angew. Chem. Int. Ed. Engl., 2013, 52, 2744-2792
127. Lamb, J.; Crawford, E.D.; Peck, D.; Modell, J.W.; Blat, I.C.; Wrobel, M.J.; Lerner, J.; Brunet, J.P.; Subramanian, A.; Ross, K.N.; Reich, M.; Hieronymus, H.; Wei, G.; Armstrong, S.A.; Haggarty, S.J.; Clemons, P.A.; Wei, R.; Carr, S.A.; Lander, E.S.; Golub, T.R. The Connectivity Map: using gene-expression signatures to connect small molecules, genes, and disease. Science, 2006, 313, 1929-1935
128. Keiser, M.J.; Roth, B.L.; Armbruster, B.N.; Ernsberger, P.; Irwin, J.J.; Shoichet, B.K. Relating protein pharmacology by ligand chemistry. Nat. Biotechnol., 2007, 25, 197-206
129. Campillos, M.; Kuhn, M.; Gavin, A.C.; Jensen, L.J.; Bork, P. Drug target identification using side-effect similarity. Science, 2008, 321, 263-266
130. Laggner, C.; Kokel, D.; Setola, V.; Tolia, A.; Lin, H.; Irwin, J.J.; Keiser, M.J.; Cheung, C.Y.; Minor, D.L. Jr; Roth, B.L.; Peterson, R.T.; Shoichet, B.K. Chemical informatics and target identification in a zebrafish phenotypic screen. Nat. Chem. Biol., 2011, 8, 144-146
131. Lounkine, E.; Keiser, M.J.; Whitebread, S.; Mikhailov, S.; Hamon, J.; Jenkins, J.L.; Lavan, P.; Weber, E.; Doak, A.K.; Côté, S.; Shoichet, B.K.; Urban, L. (2012). Large-scale prediction and testing of drug activity on side-effect targets. Nature, 2012, 486, 361-367
132. Fang, Y.; Li, G.; Peng, J. Optical biosensor provides insights for bradykinin B2 receptor signaling in A431 cells. FEBS Lett., 2005, 579, 6365-6374
133. Verrier, F.; An, S.; Ferrie, A.M.; Sun, H.; Kyoung, M.; Deng, H.; Fang, Y.; Benkovic, S.J. GPCRs regulate the assembly of a multienzyme complex for purine biosynthesis. Nat. Chem. Biol., 2011, 7, 909-915
134. Henstridge, C.M.; Balenga, N.A.; Schröder, R.; Karg, J.K.; Platzer, W.; Martini, L.; Arthur. S.; Penman, J.; Whistler, J.L.; Kostenis, E.; Waldhoer, M.; Irving, A.J. GPR55 ligands promote receptor coupling to multiple signalling pathways. Br. J. Pharmacol., 2010, 160, 604-614.