RESUMO
Interleukin-8 (IL8) is highly expressed by injured arteries in a variety of diseases and is a chemoattractant for neutrophils which express IL8 receptors IL8RA and RB (IL8RA/B) on their membranes. Neutrophils interact with the damaged endothelium and initiate an inflammatory cascade at the site of injury. We have generated a novel translational targeted cell therapy for acute vascular injury using adenoviral vectors to overexpress IL8RA/B and green fluorescent protein (GFP) on the surface of endothelial cells (ECs) derived from human induced pluripotent stem cells (HiPS-IL8RA/B-ECs). We hypothesize that HiPS-IL8RA/B-ECs transfused intravenously into rats with balloon injury of the carotid artery will target to the injured site and compete with neutrophils, thus inhibiting inflammation and neointima formation. Young adult male Sprague-Dawley rats underwent balloon injury of the right carotid artery and received intravenous transfusion of saline vehicle, 1.5 × 10(6) HiPS-ECs, 1.5 × 10(6) HiPS-Null-ECs, or 1.5 × 10(6) HiPS-IL8RA/B-ECs immediately after endoluminal injury. Tissue distribution of HiPS-IL8RA/B-ECs was analyzed by a novel GFP DNA qPCR method. Cytokine and chemokine expression and leukocyte infiltration were measured in injured and uninjured arteries at 24 h postinjury by ELISA and immunohistochemistry, respectively. Neointimal, medial areas, and reendothelialization were measured 14 days postinjury. HiPS-IL8RA/B-ECs homed to injured arteries, inhibited inflammatory mediator expression and inflammatory cell infiltration, accelerated reendothelialization, and attenuated neointima formation after endoluminal injury while control HiPS-ECs and HiPS-Null-ECs did not. HiPS-IL8RA/B-ECs transfused into rats with endoluminal carotid artery injury target to the injured artery and provide a novel strategy to treat vascular injury.
Assuntos
Artérias Carótidas/patologia , Lesões das Artérias Carótidas/terapia , Terapia Baseada em Transplante de Células e Tecidos/métodos , Células-Tronco Pluripotentes Induzidas/transplante , Neointima/prevenção & controle , Receptores de Interleucina-8/imunologia , Animais , Artérias Carótidas/imunologia , Lesões das Artérias Carótidas/imunologia , Lesões das Artérias Carótidas/patologia , Células Endoteliais , Ensaio de Imunoadsorção Enzimática , Proteínas de Fluorescência Verde/genética , Humanos , Imuno-Histoquímica , Células-Tronco Pluripotentes Induzidas/imunologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Inflamação , Masculino , Neointima/imunologia , Neointima/patologia , Reação em Cadeia da Polimerase , Ratos , Ratos Sprague-Dawley , Receptores de Interleucina-8/genéticaRESUMO
OBJECTIVE: Interleukin-8 (IL-8) receptors IL8RA and IL8RB (IL8RA/B) on neutrophil membranes bind to IL-8 with high affinity and play a critical role in neutrophil recruitment to sites of injury and inflammation. This study tested the hypothesis that administration of rat pulmonary arterial endothelial cells (ECs) overexpressing IL8RA/B can accelerate the adhesion of ECs to the injured lung and inhibit monocrotaline-induced pulmonary inflammation, arterial thickening and hypertension, and right ventricular hypertrophy. APPROACH AND RESULTS: The treatment groups included 10-week-old ovariectomized Sprague-Dawley rats that received subcutaneous injection of PBS (vehicle), a single injection of monocrotaline (monocrotaline alone, 60 mg/kg, SC), monocrotaline followed by intravenous transfusion of ECs transduced with the empty adenoviral vector (null-EC), and monocrotaline followed by intravenous transfusion of ECs overexpressing IL8RA/B (1.5 × 10(6) cells/rat). Two days or 4 weeks after monocrotaline treatment, endothelial nitric oxide synthase, inducible nitric oxide synthase, cytokine-induced neutrophil chemoattractant-2ß (IL-8 equivalent in rat), and monocyte chemoattractant protein-1 expression, neutrophil and macrophage infiltration into pulmonary arterioles, and arteriolar and alveolar morphology were measured by histological and immunohistochemical techniques. Proinflammatory cytokine/chemokine protein levels were measured by Multiplex rat-specific magnetic bead-based sandwich immunoassay in total lung homogenates. Transfusion of ECs overexpressing IL8RA/B significantly reduced monocrotaline-induced neutrophil infiltration and proinflammatory mediator (IL-8, monocyte chemoattractant protein-1, inducible nitric oxide synthase, cytokine-induced neutrophil chemoattractant, and macrophage inflammatory protein-2) expression in lungs and pulmonary arterioles and alveoli, pulmonary arterial pressure, and pulmonary arterial and right ventricular hypertrophy and remodeling. CONCLUSIONS: These provocative findings suggest that targeted delivery of ECs overexpressing IL8RA/B is effective in repairing the injured pulmonary vasculature.
Assuntos
Células Endoteliais/transplante , Terapia Genética/métodos , Hipertensão Pulmonar/prevenção & controle , Monocrotalina , Artéria Pulmonar/metabolismo , Receptores de Interleucina-8/biossíntese , Adenoviridae/genética , Animais , Pressão Arterial , Células Cultivadas , Quimiocina CCL2/metabolismo , Quimiocinas CXC/metabolismo , Modelos Animais de Doenças , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Hipertensão Pulmonar Primária Familiar , Feminino , Vetores Genéticos , Hipertensão Pulmonar/genética , Hipertensão Pulmonar/metabolismo , Hipertensão Pulmonar/patologia , Hipertensão Pulmonar/fisiopatologia , Hipertrofia Ventricular Direita/genética , Hipertrofia Ventricular Direita/metabolismo , Hipertrofia Ventricular Direita/fisiopatologia , Hipertrofia Ventricular Direita/prevenção & controle , Macrófagos/metabolismo , Infiltração de Neutrófilos , Neutrófilos/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Ovariectomia , Artéria Pulmonar/patologia , Artéria Pulmonar/fisiopatologia , Ratos , Ratos Sprague-Dawley , Receptores de Interleucina-8/genética , Transdução Genética , Regulação para Cima , Função Ventricular Direita , Remodelação VentricularRESUMO
Elevated brachial blood pressure (BP) in childhood tracks into adulthood. Central BP and measures of arterial stiffness, such as aortic augmentation index (AIx) and pulse wave velocity (PWV), have been associated with future cardiovascular disease. This pilot study assessed the feasibility of noninvasively measuring these parameters in preschool children and explored factors that may be associated with elevated BP in this age group. Brachial BP was measured using an electronic oscillometric unit (Dinamap PRO 100) and defined as elevated when systolic BP (SBP) and/or diastolic BP (DBP) was ≥ the 90th percentile for age, gender, and height. Central BP, AIx, and PWV were measured using applanation tonometry (SphygmoCor). C-reactive protein (CRP) was measured in serum samples. Sixteen African-American preschool children were recruited (4.4 ± 0.8 years, 69% males), 6 (38%) of whom had an elevated brachial BP (110 ± 10/69 ± 4 vs. 96 ± 8/55 ± 6 mm Hg, Cohen's d = 2.2). Children with elevated brachial BP had higher central SBP (d = 1.6) and DBP (d = 1.96) (97 ± 6/68 ± 4 vs. 85 ± 8/57 ± 6 mm Hg), AIx (d = 0.88) (31 ± 8 vs. 18 ± 16%, standardized to heart rate), and CRP (3.1 [2.3-6.3] vs. 0.1 [0.1-0.3] mg/dL, d = 2). There was no significant difference in PWV between groups (d = 0.26). CRP and SBP (Spearman r = 0.70), DBP (r = 0.68), central SBP (r = 0.58), and central DBP (r = 0.71) were positively correlated. Wide confidence intervals for the estimated effect sizes indicated a large degree of uncertainty about all estimates due to the small sample size. Noninvasive assessment of central BP and arterial stiffness is feasible in preschool children. Vascular inflammation may be an important factor that influences BP at an early age. Further studies in preschool children are needed to elucidate mechanisms of early onset hypertension.