Cardiac mesenchymal cells from diabetic mice are ineffective for cell therapy-mediated myocardial repair

Abstract

Although cell therapy improves cardiac function after myocardial infarction, highly variable results and limited understanding of the underlying mechanisms preclude its clinical translation. Because many heart failure patients are diabetic, we examined how diabetic conditions affect the characteristics of cardiac mesenchymal cells (CMC) and their ability to promote myocardial repair in mice. To examine how diabetes affects CMC function, we isolated CMCs from non-diabetic C57BL/6J (CMCWT) or diabetic B6.BKS(D)-Leprdb/J (CMCdb/db) mice. When CMCs were grown in 17.5 mM glucose, CMCdb/db cells showed > twofold higher glycolytic activity and a threefold higher expression of Pfkfb3 compared with CMCWT cells; however, culture of CMCdb/db cells in 5.5 mM glucose led to metabolic remodeling characterized by normalization of metabolism, a higher NAD+/NADH ratio, and a sixfold upregulation of Sirt1. These changes were associated with altered extracellular vesicle miRNA content as well as proliferation and cytotoxicity parameters comparable to CMCWT cells. To test whether this metabolic improvement of CMCdb/db cells renders them suitable for cell therapy, we cultured CMCWT or CMCdb/db cells in 5.5 mM glucose and then injected them into infarcted hearts of non-diabetic mice (CMCWT, n = 17; CMCdb/db, n = 13; Veh, n = 14). Hemodynamic measurements performed 35 days after transplantation showed that, despite normalization of their properties in vitro, and unlike CMCWT cells, CMCdb/db cells did not improve load-dependent and -independent parameters of left ventricular function. These results suggest that diabetes adversely affects the reparative capacity of CMCs and that modulating CMC characteristics via culture in lower glucose does not render them efficacious for cell therapy.

Resveratrol attenuates cerebral ischaemia reperfusion injury via modulating mitochondrial dynamics homeostasis and activating AMPK‐Mfn1 pathway

“…As shown in Figure D‐F, compared to the control group, HR injury reduced the activity of the mitochondrial respiratory complex, and their phenotypic alterations could be reversed by resveratrol at high dose. This information indicates that resveratrol sustains mitochondrial respiratory function in the setting of reperfusion injury . In addition to mitochondrial respiratory function, we also measured glucose uptake and lactic acid production.…”

“…This information indicates that resveratrol sustains mitochondrial respiratory function in the setting of reperfusion injury. 61,62 In addition to mitochondrial respiratory function, we also measured glucose uptake and lactic acid production. As shown in Figure 3C-H, compared to the control group, HR injury reduced glucose uptake and inhibited lactic acid production, suggestive of inhibition of glucose metabolism.…”

PTEN inhibition attenuates endothelial cell apoptosis in coronary heart disease via modulating the AMPK–CREB–Mfn2‐mitophagy signaling pathway

“…After three washes with PBS, the cells were incubated with goat anti-rabbit IgG (BA1105, Boster, or A0516; Beyotime) for 1 hr. After washing, DAPI was added to stain the cell nuclei (Mehra et al, 2018).…”

Echocardiography-guided percutaneous left ventricular intracavitary injection as a cell delivery approach in infarcted mice

“…Adult, female, C57BL6/J mice were subjected to 60 min of regional myocardial ischemia followed by reperfusion as previously described[15, 17, 18]. In this model, an 8–0 nylon suture was passed under left coronary artery (LCA) approximately 2 mm below the left auricle and a nontraumatic balloon occluder was applied on the artery.…”

“…CMCs[15, 16, 18, 19] and CPCs[7, 8] were isolated and cultured as previously described. All cells were from adult male mice with C57BL/6J background, some were from GFP or RFP transgenic male mice.…”