Modulating the dysregulated migration of pulmonary arterial hypertensive smooth muscle cells with motif mimicking cell permeable peptides.
Migration of vascular smooth muscle cells is a key element in remodeling during pulmonary arterial hypertension (PAH). We are observing key alterations in the migratory characteristics of human pulmonary artery smooth muscle cells (HPASMC) isolated from transplanted lungs of subjects with PAH. Using wound migration and barrier removal assays, we demonstrate that the PAH cells migrate under quiescent growth conditions and in the absence of pro-migratory factors such as platelet derived growth factor (PDGF). Under the same conditions, in the absence of PDGF, non-PAH HPASMC show negligible migration. The dysregulated migration initiates, in part, through phosphorylation events signaled through the unstimulated PDGF receptor via focal adhesion kinase (FAK) whose total basal expression and phosphorylation at tyrosine 391 is markedly increased in the PAH cells and is inhibited by a motif mimicking cell-permeable peptide (MMCPP) targeting the Tyr751 region of the PDGF receptor and by imatinib. However, exposure of the PAH cells to PDGF further promotes migration. Inhibition of p21 activated kinases (PAK), LIM kinases (LIMK), c-Jun N-terminal kinases (JNK) and p38 mitogen-activated protein kinases (MAPK) reduces both the dysregulated and the PDGF-stimulated migration. Immunofluorescence microscopy confirms these observations showing activated JNK and p38 MAPK at the edge of the wound but not in the rest of the culture in the PAH cells. The upstream inhibitors FAK (PF-573228) and imatinib block this activation of JNK and p38 at the edge of the site of injury and correspondingly inhibit migration. MMCPP which inhibit the activation of downstream effectors of migration, cofilin and caldesmon, also limit the dysregulated migration. These results highlight key pathways which point to potential targets for future therapies of pulmonary hypertension with MMCPP.
Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts.
Jamie L
JL
Wilson
Department of Chemistry, Dartmouth College, Hanover, New Hampshire 03755.
Chamila
C
Rupasinghe
Department of Surgery, Warren Alpert Medical School, Brown University, Providence, Rhode Island 02903.
Anny
A
Usheva
Tupper Research Institute and Pulmonary, Critical Care, and Sleep Division, Tufts Medical Center, Boston, Massachusetts 02111, USA.
Rod
R
Warburton
Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts.
Chloe
C
Kaplan
Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts.
Linda
L
Taylor
Tupper Research Institute and Pulmonary, Critical Care, and Sleep Division, Tufts Medical Center, Boston, Massachusetts 02111, USA.
Nicholas
N
Hill
Department of Chemistry, Dartmouth College, Hanover, New Hampshire 03755.
Dale F
DF
Mierke
Tupper Research Institute and Pulmonary, Critical Care, and Sleep Division, Tufts Medical Center, Boston, Massachusetts 02111, USA.
Peter
P
Polgar
c-Jun N-terminal kinases (JNK); cell migration; p38 MAPK; platelet-derived growth factor (PDGF); pulmonary arterial hypertension; vascular smooth muscle cells