Enhanced anti-inflammatory effects of mesenchymal stromal cells mediated by the transient ectopic expression of CXCR4 and IL10

Abstract

Background Mesenchymal stromal cells (MSCs) constitute one of the cell types most frequently used in cell therapy. Although several studies have shown the efficacy of these cells to modulate inflammation in different animal models, the results obtained in human clinical trials have been more modest. Here, we aimed at improving the therapeutic properties of MSCs by inducing a transient expression of two molecules that could enhance two different properties of these cells. With the purpose of improving MSC migration towards inflamed sites, we induced a transient expression of the C-X-C chemokine receptor type 4 (CXCR4). Additionally, to augment the anti-inflammatory properties of MSCs, a transient expression of the anti-inflammatory cytokine, interleukin 10 (IL10), was also induced. Methods Human adipose tissue-derived MSCs were transfected with messenger RNAs carrying the codon-optimized versions of CXCR4 and/or IL10. mRNA-transfected MSCs were then studied, first to evaluate whether the characteristic phenotype of MSCs was modified. Additionally, in vitro and also in vivo studies in an LPS-induced inflamed pad model were conducted to evaluate the impact associated to the transient expression of CXCR4 and/or IL10 in MSCs. Results Transfection of MSCs with CXCR4 and/or IL10 mRNAs induced a transient expression of these molecules without modifying the characteristic phenotype of MSCs. In vitro studies then revealed that the ectopic expression of CXCR4 significantly enhanced the migration of MSCs towards SDF-1, while an increased immunosuppression was associated with the ectopic expression of IL10. Finally, in vivo experiments showed that the co-expression of CXCR4 and IL10 increased the homing of MSCs into inflamed pads and induced an enhanced anti-inflammatory effect, compared to wild-type MSCs. Conclusions Our results demonstrate that the transient co-expression of CXCR4 and IL10 enhances the therapeutic potential of MSCs in a local inflammation mouse model, suggesting that these mRNA-modified cells may constitute a new step in the development of more efficient cell therapies for the treatment of inflammatory diseases.

Co-administration of human MSC overexpressing HIF-1α increases human CD34+ cell engraftment in vivo

“…HIF-1α overexpression in MSC is not the only modification that has been implemented to enhance MSC properties in different experimental scenarios. For instance, it has been shown that modification of MSC by overexpression of key molecules involved in homing and immunosuppression (as CXCR4 and IL10) or by exofucosylation can improve MSC properties [ 28 – 30 ]. Preconditioning of these cells with pro-inflammatory cytokines, irradiation, pharmacological or chemical agents may have similar effects [ 31 – 33 ].…”

Mesenchymal Stem/Stromal Cells Overexpressing CXCR4R334X Revealed Enhanced Migration: A Lesson Learned from the Pathogenesis of WHIM Syndrome

“…Some reported functional improvements in different preclinical conditions, include transplantation 52 – 54 , myocardial infarction 55 , 56 , osteoporosis 57 , cerebral ischemia 58 , graft versus host disease (GVHD) 59 , acute lung injury 60 , diabetic retinopathy 61 , and colitis-associated tumorigenesis 62 models. In addition to stable overexpression of CXCR4, Hervás-Salcedo et al showed that mRNA-transfected MSCs also possess enhanced transient CXCR4 expression and increased migration towards SDF-1 in an LPS-induced inflamed mouse model 63 . Our results regarding the comparison of Wild-type CXCR4 overexpressing MSCs with unmanipulated MSCs are in line with their results.…”

“…Upon successful animal model studies, recent gene-editing methods could also be employed to accelerate the translational process, including AAV-mediated gene therapy, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered interspaced short palindromic repeats (CRISPR) in combination with CRISPR-associated protein (Cas) 70 . Another considerable strategy to facilitate the clinical application could be transiently expressing Mutant CXCR4 , using other safer approaches such as in vitro transcription 63 . As a prominent cell-free method, extracellular vesicles (EVs) could also attenuate the potential risk of genetic engineering and pave the way towards the clinic.…”

Mesenchymal Stem Cell-Derived Extracellular Vesicles: A Potential Therapeutic Strategy for Acute Kidney Injury

“…In addition to their anti-inflammatory and vascular-support effects, the homing ability of MSCs supplements their paracrine function, and is involved in protecting microvessel density ( 32 ). Transplanted MSCs detect signals from injured kidney cells and are chemoattracted to the damaged site ( 33 ). During AKI, endothelial cells express high levels of tumor necrosis factor (TNF) and interleukins (ILs), which can up-regulate the β subunit of very late antigen-4 and vascular adhesion molecule 1 to mediate the effect of bone marrow MSCs on endothelial cell adhesion ( 33 ).…”

“…Transplanted MSCs detect signals from injured kidney cells and are chemoattracted to the damaged site ( 33 ). During AKI, endothelial cells express high levels of tumor necrosis factor (TNF) and interleukins (ILs), which can up-regulate the β subunit of very late antigen-4 and vascular adhesion molecule 1 to mediate the effect of bone marrow MSCs on endothelial cell adhesion ( 33 ). CXC motif chemokine receptor 4 (CXCR4) is a specific receptor for chemokine stromal cell-derived factor-1 (CXCL12), and CXCR4 cells are responsible for the renal repair function in AKI ( 34 ).…”