The incorporation of extracellular vesicles from MSC into HSC increase their clonogenic capacity and engraftment ability
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PROVIDER: PRJNA494576 | ENA |
REPOSITORIES: ENA
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Project description:Treatment of graft failure after allogeneic stem cell transplantation with mesenchymal stromal cells (MSC) is being evaluated clinically. MSC can exert their functions by the release of extracellular vesicles (EV). Our aim is to analyze changes induced in hematopoietic stem cells (HSC) after the incorporation of MSC-EV. MSC were isolated from healthy donors. MSC-EV were isolated by ultracentrifugation and characterized by flow cytometry (FC), Western-blot, electron microscopy and Nanosight. EV incorporation into HSC was analyzed by FC and confocal microscopy. To study HSC gene expression, RT-PCR and arrays were performed. Apoptosis and cell cycle were evaluated by FC and clonal growth by clonogenic assays in methylcellulose. The engraftment capacity of HSC was analyzed four weeks after transplantation in NOD-SCID mice. Our results showed that MSC-EV presented the characteristic morphology, size and inmunophenotype and were able to be incorporated into HSC. MSC-EV incorporation induced an a down-regulation of pro-apoptotic genes, an overexpression of genes involved in colony formation and cell proliferation together with an activation of the JAK-STAT pathway in HSC. A significant decrease in apoptosis (p=0.001), an increase of the percentage of cells in phase S and an increased CD44 expression (p=0.013) were confirmed by FC in HSC with MSC-EV. In addition, these HSC displayed a higher CFU-GM clonogenic potential (p=0.010). Finally, in the xenotransplantation model, the engraftment ability of human HSC with MSC-EV was significantly higher (p= 0.027). In summary, the incorporation of MSC-EV induces genomic and functional changes in HSC, increasing their clonogenic capacity and their engraftment ability.
2020-10-31 | GSE120803 | GEO
Project description:The bone marrow harbours multipotent mesenchymal stromal cells (MSCs) that nurture haematopoietic stem cells (HSCs). The extracellular matrix (ECM) is an integral part of the bone marrow, and the aim of this study was therefore to (a) examine the effect of engineered ECM substrates on MSC gene expression over time, and (b) to determine the functional ability of ECM-cultured MSCs to support HSCs. ECMs were surface immobilised using films of maleic anhydride to covalently immobilise tropocollagen or fibrillar collagen type I to the substrate. Where indicated, collagen type 1 fibrils were supplemented with heparin or hyaluronic acid. All surfaces maintained MSC viability, though cell expansion was ECM dependent. Microarray analysis indicated that both time in culture and culture substrate were important factors regulating gene expression. Surprisingly, a number of MSC genes critical for the support of HSC self-renewal were down regulated during the study period. The functional effects of the different substrates were quantitatively assessed by determining cobblestone area-forming cell frequencies and colony-forming units (CFUs). Progenitor frequencies were similar for all substrates during the first 3 weeks in culture, but thereafter the reference material, plasma-treated polystyrene (PTP), outperformed ECM-based culture substrates, as predicted from expression studies. Likewise, PTP was able to maintain CFUs for extended periods, whereas HSCs rapidly exhausted themselves when cultured on any of the ECM substrates tested. The ability to regulate the expression of stromal factors using reconstituted ECM is exciting and warrants further studies to identify the ECM components/combinations that maximise the expansion of clonogenic HSCs. Keywords: time course, human MSC, extracellular matrix
2009-08-01 | GSE12635 | GEO
Project description:hBM-MSC were expanded from p2 until p10 and monitored by FACS, proliferation kinetics and differentiation ability;õA analysis were performed p2/p5 (n=10)and p5/p10 (n=5) to examine if the genetic background of hBM-MSC is changeing during their high-grade expansion;
2007-06-09 | E-MARS-6 | biostudies-arrayexpress
Project description:hBM-MSC were expanded from p2 until p10 and monitored by FACS, proliferation kinetics and differentiation ability;õA analysis were performed p2/p5 (n=10)and p5/p10 (n=5) to examine if the genetic background of hBM-MSC is changeing during their high-grade expansion;
2007-06-09 | E-MARS-5 | biostudies-arrayexpress
Project description:Human mesenchymal stem cells (hMSC) play an important role in the maintenance of bone and blood. Most protocols rely on their in vitro expansion prior to clinical use. However, several groups including our own have shown that hMSC lose proliferation and differentiation ability with serial passage in culture, limiting their clinical applications. Here we show that targeting prion protein (PrP) by chemical intervention delays this process. When PrP expression was knocked down, cultures showed significant reduction in proliferation and differentiation capacity. In contrast hMSC expanded in the presence of small molecule modulator of PrP expression, 3/689, showed extended lifespan up to 10 population doublings. Upon re-plating cultures contained more than double the number of clonogenic progenitors and showed a 10 fold increase in engraftment levels in bone marrow 5 weeks post-transplant. Human MSC treated with 3/689 showed enhanced protection from DNA damage and enhanced cell cycle progression. Gene expression profiling revealed upregulation of superoxide dismutase 2 in cells treated with 3/689, dependent on PrP expression, and suggests increased scavenging of reactive oxygen species as mechanism of action. These data point to PrP as a good target to delay loss of proliferation and differentiation of hMSC with expansion in culture. Gene expression changes in hMSC expanded in the presence of 10uM small molecule modulator of prion protein 3/689 or DMSO were analysed at passage 2 and passage 8 from 3 separate donors.
2012-01-30 | E-GEOD-31205 | biostudies-arrayexpress
Project description:Human mesenchymal stem cells (hMSC) play an important role in the maintenance of bone and blood. Most protocols rely on their in vitro expansion prior to clinical use. However, several groups including our own have shown that hMSC lose proliferation and differentiation ability with serial passage in culture, limiting their clinical applications. Here we show that targeting prion protein (PrP) by chemical intervention delays this process. When PrP expression was knocked down, cultures showed significant reduction in proliferation and differentiation capacity. In contrast hMSC expanded in the presence of small molecule modulator of PrP expression, 3/689, showed extended lifespan up to 10 population doublings. Upon re-plating cultures contained more than double the number of clonogenic progenitors and showed a 10 fold increase in engraftment levels in bone marrow 5 weeks post-transplant. Human MSC treated with 3/689 showed enhanced protection from DNA damage and enhanced cell cycle progression. Gene expression profiling revealed upregulation of superoxide dismutase 2 in cells treated with 3/689, dependent on PrP expression, and suggests increased scavenging of reactive oxygen species as mechanism of action. These data point to PrP as a good target to delay loss of proliferation and differentiation of hMSC with expansion in culture.
2012-01-30 | GSE31205 | GEO
Project description:Cardiac resident c-kit+ cells aroused much interest among the cardiologists at the beginning of the century. These cells are selfrenewing, clonogenic, and multipotent and possess the potential to differentiate into all cardiovascular lineages. However, one of the critical problems faced by c-kit+ cells is the poor engraftment and survival after transplantation. Therefore, researchers are focusing on developing next generation CSC products that will enhance the engraftment, survival, and regenerative capabilities of these cells. Considering this, we made a proposal that cardiac stem cells preconditioned with exosomes derived from bone marrow mesenchymal stem cells could survive and function better in a myocardial infarction model.
2015-10-01 | GSE73598 | GEO
Project description:Tyrosine kinase inhibitors (TKI) are highly effective in treatment of chronic myeloid leukemia (CML) but do not eliminate leukemia stem cells (LSC), which remain a potential source of relapse. TKI treatment effectively inhibits BCR-ABL kinase activity in CML LSC, suggesting that additional kinase-independent mechanisms contribute to LSC preservation. We investigated whether signals from the bone marrow (BM) microenvironment protect CML LSC from TKI treatment. Coculture with human BM mesenchymal stromal cells (MSC) significantly inhibited apoptosis and preserved CML stem/progenitor cells following TKI exposure, maintaining colony forming ability and engraftment potential in immunodeficient mice. We found that the N-Cadherin receptor plays an important role in MSC-mediated protection of CML progenitors from TKI. N-Cadherin-mediated adhesion to MSC was associated with increased cytoplasmic N-Cadherin-β-catenin complex formation, as well as enhanced β-catenin nuclear translocation and transcriptional activity. Increased exogenous Wnt-mediated β-catenin signaling played an important role in MSC-mediated protection of CML progenitors from TKI treatment. Our results reveal a close interplay between N-Cadherin and the Wnt-β-catenin pathway in protecting CML LSC during TKI treatment. Importantly, these results reveal novel mechanisms of resistance of CML LSC to TKI treatment, and suggest new targets for treatment designed to eradicate residual LSC in CML patients.
2013-01-01 | GSE43225 | GEO
Project description:Therapeutic use of mesangial stem cells (MSCs) for tissue repair, including heart, has great potential. MSCs from multiple sources, including those derived from human umbilical matrix, namely Wharton's jelly, may serve as a resourceful candidate. Even after more than a decade, low engraftment efficacy of MSCs, in vivo, remains a limitation which requires in-depth understanding of the mechanisms and factors involved to facilitate a better and clinically relevant outcome. We recently hypothesized and demonstrated that the sex hormone – 17-beta estradiol (E2) – facilitates protective processes within the cardiovascular system by modulating MSC function, in vitro. Here, using a proteomic approach, we investigated the angiogenic potential of MSCs in vivo and the modulatory actions of E2 on mechanisms involved in tissue repair (potentially post-ischemic injury). Employing an in vivo approach with an ovariectomized (OVX) mice model, we evaluated the effects of E2 on MSC-induced angiogenesis using Matrigel plug assay.
2022-02-17 | PXD026380 | Pride
Project description:Engraftment and differentiation of donor hematopoietic stem cells is decisive for the clinical success of allogeneic stem cell transplantation (alloSCT) and depends on the recipient’s bone marrow (BM) niche. Presumably, a damaged niche contributes to poor graft function post-alloSCT, but the underlying mechanisms and specifically the role of BM multipotent mesenchymal stroma cells (MSC) are ill defined. Upon multivariate analysis in a cohort of 732 individuals, we observed a reduced presence of proliferation-capable MSC in BM-aspirates from patients (N=196) who had undergone alloSCT (OR 0.61, p=.028). This was confirmed by paired analysis in 30 patients showing a higher frequency of samples with a lack of MSC presence post-alloSCT compared to pre-alloSCT. This reduced MSC presence was associated with reduced survival of patients following alloSCT and specifically with impaired graft function. Post-alloSCT MSC showed diminished in vitro proliferation along with a transcriptional antiproliferative signature, upregulation of epithelial-mesenchymal transition and extracellular matrix pathways and abnormal cytokine release in coculture with hematopoietic cells. In order to avoid in-vitro-culture bias we isolated the CD146+/CD45-/HLA-DR- BM cell fraction, which comprised the entire MSC population. The post-alloSCT isolated native CD146+MSC showed a similar reduction in proliferation capacity and shared the same antiproliferative transcriptomic signature as for post-alloSCT adherence-isolated MSC. Taken together, our data show that alloSCT confers damage to the proliferative capacity of native MSC, which is associated with reduced patient survival after alloSCT and impaired engraftment of allogeneic hematopoiesis. These data represent the basis to elucidate mechanisms of BM-niche reconstitution post-alloSCT and its therapeutic manipulation.
2023-05-01 | GSE203246 | GEO