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Functional Proteomics for the Characterization of Impaired Cellular Responses to Glucocorticoids in Asthma

  • Konrad PazdrakEmail author
  • Alexander Kurosky
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 795)

Abstract

In chronic airway inflammatory disorders, such as asthma, glucocorticoid (GC) insensitivity is a challenging clinical problem associated with life-threatening disease progression and the potential development of serious side effects. The mechanism of steroid resistance in asthma remains unclear and may be multifactorial. Excluding noncompliance with GC treatment, abnormal steroid pharmacokinetics, and rare genetic defects in the glucocorticoid receptor (GR), the majority of GC insensitivity in asthma can be attributed to secondary defects related to GR function. Airway inflammatory cells obtained from patients with GC-resistant asthma show a number of abnormalities in cell immune responses to GC, which suggests that there is a causative defect in GR signaling in GC-resistant cells that could be further elucidated by a functional and molecular proteomics approach. Since T cells, eosinophils, and monocytes play a major role in the pathogenesis of airway inflammation, most of the work published to date has focused on these cell types as the primary therapeutic targets in GC-insensitive asthma. We herein review several distinct techniques for the assessment of (1) the cellular response to GCs including the effect of GCs on cell viability, adhesion, and mediator release; (2) the functionality of GC receptors, including phosphorylation of the GR, nuclear translocation, and binding activities; and (3) the characterization of proteins differentially expressed in steroid-resistant cells by comparative 2DE-gel electrophoresis-based techniques and mass spectrometry. These comprehensive approaches are expected to reveal novel candidates for biomarkers of steroid insensitivity, which may lead to the development of effective therapeutic interventions for patients with chronic steroid-resistant asthma.

Keywords

Steroid resistance Glucocorticoids Proteomics Asthma Mass spectrometry Cell signaling pathways Biomarkers Eosinophils Airway inflammatory cells Phosphoproteomics 

Abbreviations

7-AAD

7-Aminoactinomycin D

2DE

2-Dimensional electrophoresis

ADAM

A Disintegrin and metalloprotease domain family gene/protein

ASK1

Apoptosis signal-regulating kinase 1

CCL

C–C motif ligand

Cdk

Cyclin-dependent kinase

COPD

Chronic obstructive pulmonary disease

GC

Glucocorticoid

GR

Glucocorticoid receptor

GRE

Glucocorticoid response element

HDAC

Histone deacetylase

HPLC

High-performance liquid chromatography

HSP90

Heat-shock protein 90

IFN

Interferon

IL

Interleukin

JNK

c-JUN N-terminal kinase

LPS

Lipopolysaccharide

MKP-1

MAP kinase phosphatase 1

MS

Mass spectrometry

PBMC

Peripheral blood mononuclear cells

PE

Phycoerythrin

PP5

Protein phosphatase 5

T2M

Type 2 myeloid cell

TNFα

Tumor necrosis factor α

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular Biology, Institute for Translational SciencesNHLBI Proteomics Center for Airway Inflammation, University of Texas Medical BranchGalvestonUSA
  2. 2.Department of Biochemistry and Molecular Biology, Institute for Translational SciencesNHLBI Proteomics Center for Airway Inflammation, University of Texas Medical BranchGalvestonUSA

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