title: CXCR4 regulates migration of lung alveolar epithelial cells through activation of Rac1 and matrix metalloproteinase-2.
abstract: Restoration of the epithelial barrier following acute lung injury is critical for recovery of lung homeostasis. After injury, alveolar type II epithelial (ATII) cells spread and migrate to cover the denuded surface and, eventually, proliferate and differentiate into type I cells. The chemokine CXCL12, also known as stromal cell-derived factor 1α, has well-recognized roles in organogenesis, hematopoiesis, and immune responses through its binding to the chemokine receptor CXCR4. While CXCL12/CXCR4 signaling is known to be important in immune cell migration, the role of this chemokine-receptor interaction has not been studied in alveolar epithelial repair mechanisms. In this study, we demonstrated that secretion of CXCL12 was increased in the bronchoalveolar lavage of rats ventilated with an injurious tidal volume (25 ml/kg). We also found that CXCL12 secretion was increased by primary rat ATII cells and a mouse alveolar epithelial (MLE12) cell line following scratch wounding and that both types of cells express CXCR4. CXCL12 significantly increased ATII cell migration in a scratch-wound assay. When we treated cells with a specific antagonist for CXCR4, AMD-3100, cell migration was significantly inhibited. Knockdown of CXCR4 by short hairpin RNA (shRNA) caused decreased cell migration compared with cells expressing a nonspecific shRNA. Treatment with AMD-3100 decreased matrix metalloproteinase-14 expression, increased tissue inhibitor of metalloproteinase-3 expression, decreased matrix metalloproteinase-2 activity, and prevented CXCL12-induced Rac1 activation. Similar results were obtained with shRNA knockdown of CXCR4. These findings may help identify a therapeutic target for augmenting epithelial repair following acute lung injury.
authors.affiliation: Department of Physiology, Univ. of Tennessee Health Science Center, Memphis, TN 38163, USA.
authors.forename: Manik C
authors.initials: MC
authors.lastname: Ghosh
authors.forename: Patrudu S
authors.initials: PS
authors.lastname: Makena
authors.forename: Vijay
authors.initials: V
authors.lastname: Gorantla
authors.forename: Scott E
authors.initials: SE
authors.lastname: Sinclair
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Waters
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D055371:Acute Lung Injury / Q000378:metabolism / Q000473:pathology; D000818:Animals; D001596:Benzylamines; D002465:Cell Movement; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000378:metabolism / Q000502:physiology; D000080027:Cyclams; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000378:metabolism / Q000502:physiology*; D055785:Gene Knockdown Techniques; D006571:Heterocyclic Compounds / Q000494:pharmacology; D008297:Male; D053511:Matrix Metalloproteinase 14 / Q000235:genetics / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D051379:Mice; D061251:Primary Cell Culture; D011650:Pulmonary Alveoli / Q000473:pathology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D020830:rac1 GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D001596:Benzylamines; D054377:Chemokine CXCL12; C486264:Cxcr4 protein, rat; D000080027:Cyclams; D004791:Enzyme Inhibitors; D006571:Heterocyclic Compounds; D019718:Receptors, CXCR4; D019717:Tissue Inhibitor of Metalloproteinase-3; D020778:Matrix Metalloproteinase 2; C522363:Mmp2 protein, rat; D053511:Matrix Metalloproteinase 14; C499134:Rac1 protein, rat; D020830:rac1 GTP-Binding Protein; C088327:plerixafor