Obesity reduces the pro-angiogenic potential of adipose tissue stem cell-derived extracellular vesicles (EVs) by impairing miR-126 content: impact on clinical applications

Abstract

Background/Objectives:Soluble factors and cell-derived extracellular vesicles (EVs) are crucial tissue repair mediators in cell-based therapy. In the present study, we investigate the therapeutic impact of EVs released by adipose tissue-derived stem cells (ASCs) recovered from obese subjects' visceral and subcutaneous tissues.Methods:ASCs were recovered from 10 obese (oASCs) and 6 non-obese (nASCs) participants and characterized. In selected experiments, nASCs and oASCs were cultured with palmitic acid (PA) or high glucose (HG), respectively. EVs from obese (oEVs) and non-obese (nEVs) subjects' visceral and subcutaneous ASCs were collected after ultracentrifugation and analyzed for their cargo: microRNA-126 (miR-126), vascular endothelial growth factor (VEGF), and matrix metalloproteinase 2 (MMP-2), and for their biological effects on endothelial cells (ECs). Western blotting analysis and loss- and gain-of function experiments were performed.Results:oEVs show impaired angiogenic potential compared with nEVs. This effect depends on EV cargo: reduced content of VEGF, MMP-2 and, more importantly, miR-126. We demonstrate, using gain- and loss-of-function experiments, that this reduced miR-126 content leads to Spred1 upregulation and the inhibition of the extracellular signal–regulated kinase 1/2 mitogen-activated protein kinase pathway in ECs. We also show that PA treatment of nASCs translates into the release of EVs that recapitulate oEV cargo. Moreover, HG treatment of oASCs further reduces miR-126 EV content and EV-mediated in vitro angiogenesis. Finally, impaired pro-angiogenic potential is also detected in EVs released from obese subcutaneous adipose tissue-derived ASCs.Conclusions:These results indicate that obesity impacts on EV pro-angiogenic potential and may raise concerns about the use of adipose tissue-derived EVs in cell-based therapy in the obese setting.

Protective Role of Stem Cell Derived Extracellular Vesicles in an In Vitro Model of Hyperglycemia-Induced Endothelial Injury

“…This confirms that ASC and MSC EVs have both pro-angiogenic properties [11,24] and are able to restore tube formation ability in endothelial cells damaged by high glucose conditioning. These biological effects are consistent with previous observation that MSC EVs and ASC EVs contain several proangiogenic factors [11][12][13][14][24][25][26][27] and miRNAs [20][21][22]46]. …”

“…Moreover, they carry matrix metalloproteinases (MMPs) that play an important role in angiogenesis by facilitating endothelial cell migration and by promoting activation of angiogenic growth factors and other signaling molecules [25]. It has been also shown that the proangiogenic content of ASC EVs is modulated by the microenvironment, for example in vitro by the addition of different growth factors [26], or in vivo by obesity, which reduces their pro-angiogenic potential [27].…”

Insight into the development of obesity: functional alterations of adipose‐derived mesenchymal stem cells

“…In a mouse model, obese ASCs reduced their ability to form capillarylike branch structures (97). Recently, it has been shown that EVs derived from obese ASCs display impaired angiogenic potential compared with EVs from non-obese ASCs (140). Further analysis demonstrates that angiogenic-related factors like VEGF and miR-126 were reduced in obese ASC EVs, leading to the decreased activation of the Erk1/2 pathway, critical for the VEGF induced angiogenesis transduction (140).…”

MicroRNAs as paracrine signaling mediators in cancers and metabolic diseases

“…These EV mRNAs and miRNAs could target several pathways such as angiogenesis, cellular transport, or apoptosis [36]. In contrast, reduced levels of vascular endothelial growth factor (VEGF), Matrix metalloproteinase-2 (MMP-2) and especially miR-126 can impair the angiogenic potential of EV in obesity [37]. Alterations in miR-126 levels can deregulate Sprouty-related, EVH1 domain-containing 1 (Spred1) expression and inhibit the Erk1/2 mitogen-activated protein kinase (MAPK) pathway in endothelial cells and impaired angiogenic ability.…”