Selective intrarenal delivery of mesenchymal stem cell-derived extracellular vesicles attenuates myocardial injury in experimental metabolic renovascular disease

Abstract

Background: Extracellular vesicles (EVs) deliver genes and proteins to recipient cells, and mediate paracrine actions of their parent cells. Intrarenal delivery of mesenchymal stem cell (MSC)-derived EVs preserves stenotic-kidney function and reduces release of pro-inflammatory cytokines in a swine model of coexisting metabolic syndrome (MetS) and renal artery stenosis (RAS). We hypothesized that this approach is also capable of blunting cardiac injury and dysfunction.Methods: Five groups of pigs were studied after 16 weeks of diet-induced MetS and RAS (MetS +RAS), MetS and MetS+RAS treated 4 weeks earlier with a single intra-renal delivery of EVsrich fraction harvested from autologous adipose tissue-derived MSCs, and Lean and MetS Shams. Cardiac structure, function, and myocardial oxygenation were assessed in-vivo using imaging, and cardiac inflammation, senescence, and fibrosis ex-vivo. Inflammatory cytokine levels were measured in circulating and renal vein blood. Results:Intrarenal EV delivery improved stenotic-kidney glomerular filtration rate and renal blood flow, and decreased renal release of monocyte-chemoattractant protein-1 and interleukin-6. Furthermore, despite unchanged systemic hemodynamics, intrarenal EV delivery in MetS+RAS normalized cardiac diastolic function, attenuated left ventricular remodeling, cellular senescence and inflammation, and improved myocardial oxygenation and capillary density in MetS+RAS. Conclusions:Intrarenal delivery of MSC-derived EVs blunts myocardial injury in experimental MetS+RAS, possibly related to improvement in renal function and systemic inflammatory profile. These observations underscore the central role of inflammation in the crosstalk between the kidney and heart, and the important contribution of renal function to cardiac structural and functional integrity in coexisting MetS and RAS.

Metabolic syndrome increases senescence-associated micro-RNAs in extracellular vesicles derived from swine and human mesenchymal stem/stromal cells

“…For EV characterization, 12 three-months-old female domestic pigs (Manthei Hog Farm, Elk River, MN) were observed for 16 weeks in two groups. Control pigs (Lean) were fed a standard chow (13% protein, 2% fat, 6% fiber, Purina Animal Nutrition LLC, MN), and MetS pigs a metabolic-atherosclerotic diet (5B4L, protein 16.1%, ether extract fat 43.0%, and carbohydrates 40.8%, Purina Test Diet, Richmond, IN) ( n = 6 each) [ 17 ]. After completion of diet, systemic parameters including body weight, blood pressure (measured with an intra-arterial catheter), and venous levels of cholesterol, triglycerides, fasting glucose, and insulin were measured by standard procedures [ 18 ].…”

“…Abdominal subcutaneous adipose tissue collection, MSC isolation and culture, and EV collection were performed as previously described [17].…”

“…MSCs were previously characterized by the expression of CD44, CD90, and CD105, and lack of expression of CD45, CD34, CD14, using immunofluorescent staining and flow-cytometry [17]. Isolated EVs were then characterized based on the expression of EV (CD9, CD29, CD63) and MSC (CD73, CD105) markers by Western blot [17]. In human subjects, adipose tissue (5-10 g) was collected during bariatric (MetS patients) or cosmetic (Lean) surgeries.…”

“…The pig RVD model and EV injection was performed as previously described [17,23]. Twenty-four additional female domestic pigs were randomized into four groups: Lean, RVD, RVD + Lean-MSC-EVs and RVD + MetS-MSC-EVs (n = 6 each).…”

A Novel Approach to Deliver Therapeutic Extracellular Vesicles Directly into the Mouse Kidney via Its Arterial Blood Supply

“…The development of IA therapeutics for kidney disease, however, relies on the availability of animal models and feasibility of surgical techniques. Previous studies that injected MSCs directly into the renal artery have used larger animal models, including cats [23], pigs [24], rats [25], and sheep [26]. Indeed, a meta-analysis of 21 studies applying MSC therapy to animal models of renal failure has suggested that IA delivery of MSCs has greater therapeutic effect compared to intravenous delivery [27]; it should be noted though that the studies used different animal and disease models, and none of them did a direct comparison between delivery routes.…”