Tissue Kallikrein Is Essential for Invasive Capacity of Circulating Proangiogenic Cells

• Published in: Circulation research Vol. 108; no. 3; pp. 284 - 293
• Main Authors: Spinetti, Gaia, Fortunato, Orazio, Cordella, Daniela, Portararo, Paola, Kränkel, Nicolle, Katare, Rajesh, Sala-Newby, Graciela B., Richer, Christine, Vincent, Marie-Pascale, Alhenc-Gelas, Francois, Tonolo, Giancarlo, Cherchi, Sara, Emanueli, Costanza, Madeddu, Paolo
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD American Heart Association, Inc 04.02.2011; Lippincott Williams & Wilkins
• Subjects: Adult; Aged; Animals; Biological and medical sciences; Case-Control Studies; Cell Movement - physiology; Diabetes Mellitus, Type 2 - metabolism; Diabetes Mellitus, Type 2 - pathology; Down-Regulation; Female; Fundamental and applied biological sciences. Psychology; Hindlimb - blood supply; Humans; Ischemia - metabolism; Kallikreins - genetics; Kallikreins - metabolism; Kinins - metabolism; Leukocytes, Mononuclear - cytology; Leukocytes, Mononuclear - metabolism; Male; Matrix Metalloproteinase 2 - metabolism; Mice; Mice, Inbred C57BL; Mice, Knockout; Middle Aged; Models, Animal; Neovascularization, Physiologic - physiology; Nitric Oxide Synthase - metabolism; Receptor, Bradykinin B2 - metabolism; RNA, Messenger - metabolism; Signal Transduction - physiology; Vertebrates: cardiovascular system; kallikrein―kinin system; Angiogenesis; Tissue; Vertebrata; Mammalia; circulating proangiogenic cells; Kallikrein kinin system; Kallikrein; Circulatory system; cell invasion; Essential; Invasion
• ISSN: 0009-7330; 1524-4571; 1524-4571
• DOI: 10.1161/CIRCRESAHA.110.236786

RATIONALE:Homing of proangiogenic cells (PACs) is guided by chemoattractants and requires proteases to disrupt the extracellular matrix. The possibility that PAC recruitment involves an interaction between proteases and chemotactic factor receptors remains largely unexplored. OBJECTIVE:To determine the role of human tissue kallikrein (hK1) in PAC invasion and its dependency on kinin receptor signaling. METHODS AND RESULTS:Human mononuclear cells (MNCs) and culture-selected PACs express and release mature hK1 protein. HK1 gene (KLK1) silencing reduced PACs migratory, invasive, and proangiogenic activities. KLK1-knockout mouse bone marrow–derived MNCs showed similar impairments and were unable to support reparative angiogenesis in a mouse model of peripheral ischemia. Conversely, adenovirus-mediated KLK1 (Ad.KLK1) gene transfer enhanced PAC-associated functions, whereas the catalytically inactive variant R53H-KLK1 was ineffective. HK1-induced effects are mediated by a kinin B2 receptor (B2R)-dependent mechanism involving inducible nitric oxide synthase and metalloproteinase-2 (MMP2). Lower hK1 protein levels were observed in PACs from type 2 diabetic (T2D) patients, whereas KLK1 mRNA levels were similar to those of healthy subjects, suggesting a post-transcriptional defect. Furthermore, B2R is normally expressed on T2D-PACs but remains uncoupled from downstream signaling. Importantly, whereas Ad.KLK1 alone could not restore T2D-PAC invasion capacity, combined KLK1 and B2R expression rescued the diabetic phenotype. CONCLUSIONS:This study reveals new interactive components of the PACs invasive machinery, acting via protease- and kinin receptor–dependent mechanisms.