Overexpression of Nitric Oxide Synthase Restores Circulating Angiogenic Cell Function in Patients With Coronary Artery Disease: Implications for Autologous Cell Therapy for Myocardial Infarction

Abstract

BackgroundCirculating angiogenic cells (CACs) are peripheral blood cells whose functional capacity inversely correlates with cardiovascular risk and that have therapeutic benefits in animal models of cardiovascular disease. However, donor age and disease state influence the efficacy of autologous cell therapy. We sought to determine whether age or coronary artery disease (CAD) impairs the therapeutic potential of CACs for myocardial infarction (MI) and whether the use of ex vivo gene therapy to overexpress endothelial nitric oxide (NO) synthase (eNOS) overcomes these defects.Methods and ResultsWe recruited 40 volunteers varying by sex, age (< or ≥45 years), and CAD and subjected their CACs to well‐established functional tests. Age and CAD were associated with reduced CAC intrinsic migration (but not specific response to vascular endothelial growth factor, adherence of CACs to endothelial tubes, eNOS mRNA and protein levels, and NO production. To determine how CAC function influences therapeutic potential, we injected the 2 most functional and the 2 least functional CAC isolates into mouse hearts post MI. The high‐function isolates substantially improved cardiac function, whereas the low‐function isolates led to cardiac function only slightly better than vehicle control. Transduction of the worst isolate with eNOS cDNA adenovirus increased NO production, migration, and cardiac function of post‐MI mice implanted with the CACs. Transduction of the best isolate with eNOS small interfering RNA adenovirus reduced all of these capabilities.ConclusionsAge and CAD impair multiple functions of CACs and limit therapeutic potential for the treatment of MI. eNOS gene therapy in CACs from older donors or those with CAD has the potential to improve autologous cell therapy outcomes.

Chronic stress is associated with reduced circulating hematopoietic progenitor cell number: A maternal caregiving model

“…A previously published study by our lab found that psychological stress was associated with reduced migratory and paracrine function of CACs (a cell culture model containing CPCs) in vitro (19). In vitro CAC function is highly correlated with vascular repair capacity in vivo (49). The effects of stress on cell-mediated vascular repair in vivo could be tested by isolating human CACs or CD34 + cells from high versus low-stress healthy adults, and transplanting them into an animal model of vascular injury (49).…”

“…In vitro CAC function is highly correlated with vascular repair capacity in vivo (49). The effects of stress on cell-mediated vascular repair in vivo could be tested by isolating human CACs or CD34 + cells from high versus low-stress healthy adults, and transplanting them into an animal model of vascular injury (49). …”

Circulating angiogenic cell function is inhibited by cortisol in vitro and associated with psychological stress and cortisol in vivo

“…The current study provides a foundation to test stress pathways using human-to-animal CAC transplant models of vascular repair (e.g., post-myocardial infarction or leg ischemia)(Sonnenschein et al, 2011; Chen et al, 2016). For example, a previous study of patients with metabolic syndrome demonstrated that exercise improved the ability of these patients’ CACs to repair a carotid endothelial injury in vivo in a nude mouse model (Sonnenschein et al, 2011).…”

“…CAC migration in vitro reflects the capacity of CACs to migrate toward sites of tissue damage and promote repair via paracrine secretion of growth factors. CAC migration is decreased in patients with coronary artery disease (Vasa et al, 2001), atherosclerosis (Ohtsuka et al, 2013), diabetes (Thum et al, 2007), and older age (Chen et al, 2016). Among healthy individuals without cardiovascular disease or diabetes, reduced CAC migration prospectively predicts greater carotid artery intima-media thickness (Keymel et al, 2008) and correlates with metabolic risk factors (Aschbacher et al, 2012a) and better endothelial function (Van Craenenbroeck et al, 2010).…”

Associations of theNOS3rs1799983 polymorphism with circulating nitric oxide and lipid levels: a systematic review and meta-analysis

“…Among them, NOS3 is responsible for synthesising circulating NO and plays a crucial role in the pathogenesis of CAD 4 5. Chen et al 4 demonstrated that the mRNA and protein levels of NOS3 were significantly decreased in patients with CAD as compared with healthy subjects, and overexpression of NOS3 increased NO production and restored cardiac function after myocardial infarction in mice. In a mouse model of arterial restenosis, the antirestenotic effect of liraglutide was completely abolished by N-omega-nitro-L-arginine methyl ester, an inhibitor of NOS3 5…”