miR-21 modulates tumor outgrowth induced by human adipose tissue-derived mesenchymal stem cells in vivo.
ABSTRACT Mesenchymal stem cells (MSCs) have generated a great deal of interest in clinical situations, due principally to their potential use in regenerative medicine and tissue engineering applications. However, the therapeutic application of MSCs remains limited, unless the favorable effects of MSCs on tumor growth in vivo, and the long-term safety of the clinical applications of MSCs, can be more thoroughly understood. In this study, we determined whether microRNAs can modulate MSC-induced tumor outgrowth in BALB/c nude mice. Overexpression of miR-21 in human adipose-derived stem cells (hADSCs) inhibited hADSC-induced tumor growth, and inhibition of miR-21 increased it. Downregulation of transforming growth factor beta receptor II (TGFBR2), but not of signal transducer and activator of transcription 3, in hADSCs showed effects similar to those of miR-21 overexpression. Downregulation of TGFBR2 and overexpression of miR21 decreased tumor vascularity. Inhibition of miR-21 and the addition of TGF-β increased the levels of vascular endothelial growth factor and interleukin-6 in hADSCs. Transplantation of miR-21 inhibitor-transfected hADSCs increased blood flow recovery in a hind limb ischemia model of nude mice, compared with transplantation of control oligo-transfected cells. These findings indicate that MSCs might favor tumor growth in vivo. Thus, it is necessary to study the long-term safety of this technique before MSCs can be used as therapeutic tools in regenerative medicine and tissue engineering.
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ABSTRACT: Stem cells are one of the most powerful tools in regeneration medicine. However, many limitations remain regarding the use of adult stem cells in clinical applications, including poor cell survival and low treatment efficiency. We describe an innovative three-dimensional cell mass (3DCM) culture that is based on cell adhesion (basic fibroblast growth factor-immobilized substrate) and assess the therapeutic potential of 3DCMs composed of human adipose tissue-derived stromal cells (hASCs). For formation of a 3DCM, hASCs were cultured on a substrate with immobilized fibroblast growth factor-2. The angiogenic potential of 3DCMs was determined by immunostaining, fluorescence-activated cell sorting and protein analysis. To evaluate the vasculature ability and improved treatment efficacy of 3DCMs, the 3DCMs were intramuscularly injected into the ischemic limbs of mice. The 3DCMs released various angiogenic factors (eg, vascular endothelial growth factor and interleukin-8) and differentiated into vascular cells within 3 days in normal medium. Blood vessel and tissue regeneration was clearly observed through visual inspection in the 3DCM-injected group. hASC injection slowed limb necrosis after treatment, but 50% of the mice ultimately had limb loss within 28 days. Most mice receiving 3DCMs had limb salvage (89%) or mild limb necrosis (11%). 3DCM culture promotes the efficient vascular differentiation of stem cells, and 3DCM transplantation results in the direct vascular regeneration of the injected cells and an improved therapeutic efficacy.Cytotherapy 11/2013; · 3.06 Impact Factor
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ABSTRACT: Finding a suitable cell source for endothelial cells (ECs) for cardiovascular regeneration is a challenging issue for regenerative medicine. In the paper we describe a novel mechanism regulating induced pluripotent stem cells (iPSC) differentiation into ECs, with a particular focus on miRNAs and their targets. We first established a protocol using collagen IV and VEGF to drive the functional differentiation of iPSCs into ECs and compared the miRNA signature of differentiated and undifferentiated cells. Among the miRNAs overrepresented in differentiated cells, we focused on microRNA-21 (miR-21) and studied its role in iPSC differentiation. Overexpression of miR-21 in pre-differentiated iPSCs induced EC marker upregulation and in vitro and in vivo capillary formation; accordingly, inhibition of miR-21 produced the opposite effects. Importantly, miR-21 overexpression increased TGF-β2 mRNA and secreted protein level, consistent with the strong upregulation of TGF-β2 during iPSC differentiation. Indeed, treatment of iPSCs with TGFβ-2 induced EC marker expression and in vitro tube formation. Inhibition of SMAD3, a downstream effector of TGFβ-2, strongly decreased VE-cadherin expression. Furthermore, TGFβ-2 neutralization and knockdown inhibited miR-21-induced EC marker expression. Finally, we confirmed the PTEN/Akt pathway as a direct target of miR-21 and we showed that PTEN knockdown is required for miR-21 mediated endothelial differentiation. In conclusion, we elucidated a novel signaling pathway that promotes the differentiation of iPSC into functional ECs suitable for regenerative medicine applications.Journal of Biological Chemistry 12/2013; · 4.65 Impact Factor