title: A novel small molecule target in human airway smooth muscle for potential treatment of obstructive lung diseases: a staged high-throughput biophysical screening.
abstract: 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.
authors.affiliation: Division of Physiology, Department of Environmental Health Sciences, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA. san@jhsph.edu
authors.forename: Steven S
authors.initials: SS
authors.lastname: An
authors.forename: Peter S
authors.initials: PS
authors.lastname: Askovich
authors.forename: Thomas I
authors.initials: TI
authors.lastname: Zarembinski
authors.forename: Kwangmi
authors.initials: K
authors.lastname: Ahn
authors.forename: John M
authors.initials: JM
authors.lastname: Peltier
authors.forename: Moritz
authors.initials: M
authors.lastname: von Rechenberg
authors.forename: Sudhir
authors.initials: S
authors.lastname: Sahasrabudhe
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Fredberg
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism*; D000818:Animals; D001993:Bronchodilator Agents / Q000737:chemistry / Q000494:pharmacology*; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery / Q000379:methods*; D005454:Fluorescence Polarization; D005583:Fourier Analysis; D050886:HSP20 Heat-Shock Proteins / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D006801:Humans; D066298:In Vitro Techniques; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D008173:Lung Diseases, Obstructive / Q000188:drug therapy* / Q000378:metabolism; D008280:Magnetics; D008297:Male; D015394:Molecular Structure; D009126:Muscle Relaxation / Q000187:drug effects*; D009130:Muscle, Smooth / Q000187:drug effects* / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D011916:Rats, Inbred F344; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects; D054852:Small Molecule Libraries; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance; D013997:Time Factors
chemical_list: D048948:14-3-3 Proteins; D001993:Bronchodilator Agents; D050886:HSP20 Heat-Shock Proteins; D054852:Small Molecule Libraries