A role for Gab1/SHP2 in thrombin activation of PAK1

Gene transfer of kinase-dead pak1 inhibits injury-induced restenosis

Dong Wang, Biman C. Paria, Qiuhua Zhang, Manjula Karpurapu, Quanyi Li, William T. Gerthoffer, Yoshikazu Nakaoka, Rao Gadiparthi

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

To understand the role of epidermal growth factor receptor (EGFR) transactivation in G protein-coupled receptor (GPCR) agonist-induced signaling events, we have studied the capacity of thrombin in the activation of Gab1-SHP2 in vascular smooth muscle cells (VSMCs). Thrombin activated both Gab1 and SHP2 in EGFR-dependent manner. Similarly, thrombin induced Rac1 and Cdc42 activation, and these responses were suppressed when either Gab1 or SHP2 stimulation is blocked. Thrombin also induced PAK1 activation in a time- and EGFR-Gab1-SHP2-Rac1/Cdc42-dependent manner. Inhibition of activation of EGFR, Gab1, SHP2, Rac1, Cdc42, or PAK1 by pharmacological or genetic approaches attenuated thrombin-induced VSMC stress fiber formation and motility. Thrombin activated RhoA in a time-dependent manner in VSMCs. LARG, a RhoA-specific GEF (guanine nucleotide exchange factor), was found to be associated with Gab1 and siRNA-mediated depletion of its levels suppressed RhoA, Rac1 and PAK1 activation. Dominant negative mutant-mediated interference of RhoA activation inhibited thrombin-induced Rac1 and PAK1 stimulation in VSMCs and their stress fiber formation and migration. Balloon injury induced PAK1 activity and interference with its activation led to attenuation of SMC migration from media to intima, resulting in reduced neointima formation and increased lumen size. Inhibition of thrombin signaling by recombinant hirudin also blocked balloon injury-induced EGFR tyrosine phosphorylation and PAK1 activity. These results show that thrombin-mediated PAK1 activation plays a crucial role in vascular wall remodeling and it could be a potential target for drug development against these vascular lesions.

Original languageEnglish (US)
Pages (from-to)1066-1075
Number of pages10
JournalCirculation research
Volume104
Issue number9
DOIs
StatePublished - May 8 2009

Fingerprint

Thrombin
Transcriptional Activation
Phosphotransferases
Wounds and Injuries
Epidermal Growth Factor Receptor
Vascular Smooth Muscle
Smooth Muscle Myocytes
Stress Fibers
Hirudins
Guanine Nucleotide Exchange Factors
Neointima
G-Protein-Coupled Receptors
Small Interfering RNA
Blood Vessels
Tyrosine
Phosphorylation
Pharmacology
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

A role for Gab1/SHP2 in thrombin activation of PAK1 : Gene transfer of kinase-dead pak1 inhibits injury-induced restenosis. / Wang, Dong; Paria, Biman C.; Zhang, Qiuhua; Karpurapu, Manjula; Li, Quanyi; Gerthoffer, William T.; Nakaoka, Yoshikazu; Gadiparthi, Rao.

In: Circulation research, Vol. 104, No. 9, 08.05.2009, p. 1066-1075.

Research output: Contribution to journalArticle

Wang, Dong ; Paria, Biman C. ; Zhang, Qiuhua ; Karpurapu, Manjula ; Li, Quanyi ; Gerthoffer, William T. ; Nakaoka, Yoshikazu ; Gadiparthi, Rao. / A role for Gab1/SHP2 in thrombin activation of PAK1 : Gene transfer of kinase-dead pak1 inhibits injury-induced restenosis. In: Circulation research. 2009 ; Vol. 104, No. 9. pp. 1066-1075.
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abstract = "To understand the role of epidermal growth factor receptor (EGFR) transactivation in G protein-coupled receptor (GPCR) agonist-induced signaling events, we have studied the capacity of thrombin in the activation of Gab1-SHP2 in vascular smooth muscle cells (VSMCs). Thrombin activated both Gab1 and SHP2 in EGFR-dependent manner. Similarly, thrombin induced Rac1 and Cdc42 activation, and these responses were suppressed when either Gab1 or SHP2 stimulation is blocked. Thrombin also induced PAK1 activation in a time- and EGFR-Gab1-SHP2-Rac1/Cdc42-dependent manner. Inhibition of activation of EGFR, Gab1, SHP2, Rac1, Cdc42, or PAK1 by pharmacological or genetic approaches attenuated thrombin-induced VSMC stress fiber formation and motility. Thrombin activated RhoA in a time-dependent manner in VSMCs. LARG, a RhoA-specific GEF (guanine nucleotide exchange factor), was found to be associated with Gab1 and siRNA-mediated depletion of its levels suppressed RhoA, Rac1 and PAK1 activation. Dominant negative mutant-mediated interference of RhoA activation inhibited thrombin-induced Rac1 and PAK1 stimulation in VSMCs and their stress fiber formation and migration. Balloon injury induced PAK1 activity and interference with its activation led to attenuation of SMC migration from media to intima, resulting in reduced neointima formation and increased lumen size. Inhibition of thrombin signaling by recombinant hirudin also blocked balloon injury-induced EGFR tyrosine phosphorylation and PAK1 activity. These results show that thrombin-mediated PAK1 activation plays a crucial role in vascular wall remodeling and it could be a potential target for drug development against these vascular lesions.",
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