TY - JOUR
T1 - A novel small molecule target in human airway smooth muscle for potential treatment of obstructive lung diseases
T2 - A staged high-throughput biophysical screening
AU - An, Steven S.
AU - Askovich, Peter S.
AU - Zarembinski, Thomas I.
AU - Ahn, Kwangmi
AU - Peltier, John M.
AU - von Rechenberg, Moritz
AU - Sahasrabudhe, Sudhir
AU - Fredberg, Jeffrey J.
N1 - Funding Information:
This work was supported by NIH grants HL59682 (JJF) and HL33009 (JJF); by NIEHS Center grant (2P30 ES03819-11) pilot grant (SSA); and by Faculty Research Initiative Fund from Johns Hopkins Bloomberg School of Public Health (SSA).
PY - 2011/1/13
Y1 - 2011/1/13
N2 - Background: A newly identified mechanism of smooth muscle relaxation is the interaction between the small heat shock protein 20 (HSP20) and 14-3-3 proteins. Focusing upon this class of interactions, we describe here a novel drug target screening approach for treating airflow obstruction in asthma.Methods: Using a high-throughput fluorescence polarization (FP) assay, we screened a library of compounds that could act as small molecule modulators of HSP20 signals. We then applied two quantitative, cell-based biophysical methods to assess the functional efficacy of these molecules and rank-ordered their abilities to relax isolated human airway smooth muscle (ASM). Scaling up to the level of an intact tissue, we confirmed in a concentration-responsive manner the potency of the cell-based hit compounds.Results: Among 58,019 compound tested, 268 compounds caused 20% or more reduction of the polarized emission in the FP assay. A small subset of these primary screen hits, belonging to two scaffolds, caused relaxation of isolated ASM cell in vitro and attenuated active force development of intact tissue ex vivo.Conclusions: This staged biophysical screening paradigm provides proof-of-principle for high-throughput and cost-effective discovery of new small molecule therapeutic agents for obstructive lung diseases.
AB - Background: A newly identified mechanism of smooth muscle relaxation is the interaction between the small heat shock protein 20 (HSP20) and 14-3-3 proteins. Focusing upon this class of interactions, we describe here a novel drug target screening approach for treating airflow obstruction in asthma.Methods: Using a high-throughput fluorescence polarization (FP) assay, we screened a library of compounds that could act as small molecule modulators of HSP20 signals. We then applied two quantitative, cell-based biophysical methods to assess the functional efficacy of these molecules and rank-ordered their abilities to relax isolated human airway smooth muscle (ASM). Scaling up to the level of an intact tissue, we confirmed in a concentration-responsive manner the potency of the cell-based hit compounds.Results: Among 58,019 compound tested, 268 compounds caused 20% or more reduction of the polarized emission in the FP assay. A small subset of these primary screen hits, belonging to two scaffolds, caused relaxation of isolated ASM cell in vitro and attenuated active force development of intact tissue ex vivo.Conclusions: This staged biophysical screening paradigm provides proof-of-principle for high-throughput and cost-effective discovery of new small molecule therapeutic agents for obstructive lung diseases.
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U2 - 10.1186/1465-9921-12-8
DO - 10.1186/1465-9921-12-8
M3 - Article
C2 - 21232113
AN - SCOPUS:78651310497
SN - 1465-9921
VL - 12
JO - Respiratory research
JF - Respiratory research
M1 - 8
ER -