Actomyosin and the MRTF-SRF pathway downregulate FGFR1 in mesenchymal stromal cells

Abstract Both biological and mechanical signals are known to influence cell proliferation. However, biological signals are mostly studied in two-dimensions (2D) and the interplay between these different pathways is largely unstudied. Here, we investigated the influence of the cell culture environment on the response to bFGF, a widely studied and important proliferation growth factor. We observed that human mesenchymal stromal cells (hMSCs), but not fibroblasts, lose the ability to respond to soluble or covalently bound bFGF when cultured on microfibrillar substrates. This behavior correlated with a downregulation of FGF receptor 1 (FGFR1) expression of hMSCs on microfibrillar substrates. Inhibition of actomyosin or the MRTF/SRF pathway decreased FGFR1 expression in hMSCs, fibroblasts and MG63 cells. To our knowledge, this is the first time FGFR1 expression is shown to be regulated through a mechanosensitive pathway in hMSCs. These results add to the sparse literature on FGFR1 regulation and potentially aid designing tissue engineering constructs that better control cell proliferation.

Download Full-text

Proinflammatory stimuli induce galectin-9 in human mesenchymal stromal cells to suppress T-cell proliferation

European Journal of Immunology ◽

10.1002/eji.201343335 ◽

2013 ◽

Vol 43 (10) ◽

pp. 2741-2749 ◽

Cited By ~ 57

Author(s):

Friederike Gieseke ◽

Anne Kruchen ◽

Nikolay Tzaribachev ◽

Frank Bentzien ◽

Massimo Dominici ◽

...

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

T Cell Proliferation ◽

Human Mesenchymal Stromal Cells

Download Full-text

Efficient Transformation of Primary Human Mesenchymal Stromal Cells by Adenovirus Early Region 1 Oncogenes

Journal of Virology ◽

10.1128/jvi.01782-16 ◽

2016 ◽

Vol 91 (1) ◽

Cited By ~ 4

Author(s):

Thomas Speiseder ◽

Helga Hofmann-Sieber ◽

Estefanía Rodríguez ◽

Anna Schellenberg ◽

Nuray Akyüz ◽

...

Keyword(s):

Stem Cell ◽

Progenitor Cells ◽

Cell Lines ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Human Cells ◽

Target Cells ◽

Cell Type ◽

Human Mesenchymal Stromal Cells ◽

First Time

ABSTRACT Previous observations that human amniotic fluid cells (AFC) can be transformed by human adenovirus type 5 (HAdV-5) E1A/E1B oncogenes prompted us to identify the target cells in the AFC population that are susceptible to transformation. Our results demonstrate that one cell type corresponding to mesenchymal stem/stroma cells (hMSCs) can be reproducibly transformed by HAdV-5 E1A/E1B oncogenes as efficiently as primary rodent cultures. HAdV-5 E1-transformed hMSCs exhibit all properties commonly associated with a high grade of oncogenic transformation, including enhanced cell proliferation, anchorage-independent growth, increased growth rate, and high telomerase activity as well as numerical and structural chromosomal aberrations. These data confirm previous work showing that HAdV preferentially transforms cells of mesenchymal origin in rodents. More importantly, they demonstrate for the first time that human cells with stem cell characteristics can be completely transformed by HAdV oncogenes in tissue culture with high efficiency. Our findings strongly support the hypothesis that undifferentiated progenitor cells or cells with stem cell-like properties are highly susceptible targets for HAdV-mediated cell transformation and suggest that virus-associated tumors in humans may originate, at least in part, from infections of these cell types. We expect that primary hMSCs will replace the primary rodent cultures in HAdV viral transformation studies and are confident that these investigations will continue to uncover general principles of viral oncogenesis that can be extended to human DNA tumor viruses as well. IMPORTANCE It is generally believed that transformation of primary human cells with HAdV-5 E1 oncogenes is very inefficient. However, a few cell lines have been successfully transformed with HAdV-5 E1A and E1B, indicating that there is a certain cell type which is susceptible to HAdV-mediated transformation. Interestingly, all those cell lines have been derived from human embryonic tissue, albeit the exact cell type is not known yet. We show for the first time the successful transformation of primary human mesenchymal stromal cells (hMSCs) by HAdV-5 E1A and E1B. Further, we show upon HAdV-5 E1A and E1B expression that these primary progenitor cells exhibit features of tumor cells and can no longer be differentiated into the adipogenic, chondrogenic, or osteogenic lineage. Hence, primary hMSCs represent a robust and novel model system to elucidate the underlying molecular mechanisms of adenovirus-mediated transformation of multipotent human progenitor cells.

Download Full-text

Human mesenchymal stromal cells suppress T-cell proliferation independent of heme oxygenase-1

Cytotherapy ◽

10.1016/j.jcyt.2014.11.010 ◽

2015 ◽

Vol 17 (4) ◽

pp. 382-391 ◽

Cited By ~ 8

Author(s):

Seema R. Patel ◽

Ian B. Copland ◽

Marco A. Garcia ◽

Richard Metz ◽

Jacques Galipeau

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Heme Oxygenase ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

T Cell Proliferation ◽

Heme Oxygenase 1 ◽

Human Mesenchymal Stromal Cells

Download Full-text

Mechanosensitive regulation of FGFR1 through the MRTF-SRF pathway

10.1101/782243 ◽

2019 ◽

Cited By ~ 1

Author(s):

Jip Zonderland ◽

Silvia Rezzola ◽

Lorenzo Moroni

Keyword(s):

Tissue Engineering ◽

Cancer Biology ◽

Control Cell ◽

Tissue Culture Plastic ◽

Cancer Treatments ◽

Differentiation Potential ◽

Fgf Receptor ◽

Proliferation And Differentiation ◽

First Time ◽

Fgfr1 Expression

AbstractControlling basic fibroblast growth factor (bFGF) signaling is important for both tissue-engineering purposes, controlling proliferation and differentiation potential, and for cancer biology, influencing tumor progression and metastasis. Here, we observed that human mesenchymal stromal cells (hMSCs) no longer responded to soluble or covalently bound bFGF when cultured on microfibrillar substrates, while fibroblasts did. This correlated with a downregulation of FGF receptor 1 (FGFR1) expression of hMSCs on microfibrillar substrates, compared to hMSCs on conventional tissue culture plastic (TCP). hMSCs also expressed less SRF on ESP scaffolds, compared to TCP, while fibroblasts maintained high FGFR1 and SRF expression. Inhibition of actin-myosin tension or the MRTF/SRF pathway decreased FGFR1 expression in hMSCs, fibroblasts and MG63 osteosarcoma cells. This downregulation was functional, as hMSCs became irresponsive to bFGF in the presence of MRTF/SRF inhibitor. Together, our data show that hMSCs, but not fibroblasts, are irresponsive to bFGF when cultured on microfibrillar susbtrates by downregulation of FGFR1 through the MRTF/SRF pathway. This is the first time FGFR1 expression has been shown to be mechanosensitive and adds to the sparse literature on FGFR1 regulation. These results could open up new targets for cancer treatments and could aid designing tissue engineering constructs that better control cell proliferation.

Download Full-text

Impact of Tranexamic Acid on Chondrocytes and Osteogenically Differentiated Human Mesenchymal Stromal Cells (hMSCs) In Vitro

Journal of Clinical Medicine ◽

10.3390/jcm9123880 ◽

2020 ◽

Vol 9 (12) ◽

pp. 3880

Author(s):

Mike Wagenbrenner ◽

Tizian Heinz ◽

Konstantin Horas ◽

Axel Jakuscheit ◽

Joerg Arnholdt ◽

...

Keyword(s):

Cell Proliferation ◽

Tranexamic Acid ◽

Osteogenic Differentiation ◽

Mesenchymal Stromal Cells ◽

Topical Application ◽

Stromal Cells ◽

Specific Marker ◽

Human Mesenchymal Stromal Cells

The topical application of tranexamic acid (TXA) helps to prevent post-operative blood loss in total joint replacements. Despite these findings, the effects on articular and periarticular tissues remain unclear. Therefore, this in vitro study examined the effects of varying exposure times and concentrations of TXA on proliferation rates, gene expression and differentiation capacity of chondrocytes and human mesenchymal stromal cells (hMSCs), which underwent osteogenic differentiation. Chondrocytes and hMSCs were isolated and multiplied in monolayer cell cultures. Osteogenic differentiation of hMSCs was induced for 21 days using a differentiation medium containing specific growth factors. Cell proliferation was analyzed using ATP assays. Effects of TXA on cell morphology were examined via light microscopy and histological staining, while expression levels of tissue-specific genes were measured using semiquantitative RT-PCR. After treatment with 50 mg/mL of TXA, a decrease in cell proliferation rates was observed. Furthermore, treatment with concentrations of 20 mg/mL of TXA for at least 48 h led to a visible detachment of chondrocytes. TXA treatment with 50 mg/mL for at least 24 h led to a decrease in the expression of specific marker genes in chondrocytes and osteogenically differentiated hMSCs. No significant effects were observed for concentrations beyond 20 mg/mL of TXA combined with exposure times of less than 24 h. This might therefore represent a safe limit for topical application in vivo. Further research regarding in vivo conditions and effects on hMSC functionality are necessary to fully determine the effects of TXA on articular and periarticular tissues.

Download Full-text

Indoleamine 2,3-dioxygenase mediates inhibition of virus-specific CD8+ T cell proliferation by human mesenchymal stromal cells

Cytotherapy ◽

10.1016/j.jcyt.2016.01.009 ◽

2016 ◽

Vol 18 (5) ◽

pp. 621-629 ◽

Cited By ~ 9

Author(s):

Jian Hong ◽

Angela Hueckelhoven ◽

Lei Wang ◽

Anita Schmitt ◽

Patrick Wuchter ◽

...

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Cd8 T Cell ◽

T Cell Proliferation ◽

Human Mesenchymal Stromal Cells ◽

Indoleamine 2,3 Dioxygenase

Download Full-text

Multi-pronged approach to human mesenchymal stromal cells senescence quantification with a focus on label-free methods

Scientific Reports ◽

10.1038/s41598-020-79831-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Weichao Zhai ◽

Jerome Tan ◽

Tobias Russell ◽

Sixun Chen ◽

Dennis McGonagle ◽

...

Keyword(s):

Flow Cytometry ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Preclinical Model ◽

Label Free ◽

Current Standard ◽

Differentiation Potential ◽

Human Mesenchymal Stromal Cells ◽

Endogenous Fluorophores

AbstractHuman mesenchymal stromal cells (hMSCs) have demonstrated, in various preclinical settings, consistent ability in promoting tissue healing and improving outcomes in animal disease models. However, translation from the preclinical model into clinical practice has proven to be considerably more difficult. One key challenge being the inability to perform in situ assessment of the hMSCs in continuous culture, where the accumulation of the senescent cells impairs the culture’s viability, differentiation potential and ultimately leads to reduced therapeutic efficacies. Histochemical $$\upbeta $$ β -galactosidase staining is the current standard for measuring hMSC senescence, but this method is destructive and not label-free. In this study, we have investigated alternatives in quantification of hMSCs senescence, which included flow cytometry methods that are based on a combination of cell size measurements and fluorescence detection of SA-$$\upbeta $$ β -galactosidase activity using the fluorogenic substrate, C$${_{12}}$$ 12 FDG; and autofluorescence methods that measure fluorescence output from endogenous fluorophores including lipopigments. For identification of senescent cells in the hMSC batches produced, the non-destructive and label-free methods could be a better way forward as they involve minimum manipulations of the cells of interest, increasing the final output of the therapeutic-grade hMSC cultures. In this work, we have grown hMSC cultures over a period of 7 months and compared early and senescent hMSC passages using the advanced flow cytometry and autofluorescence methods, which were benchmarked with the current standard in $$\upbeta $$ β -galactosidase staining. Both the advanced methods demonstrated statistically significant values, (r = 0.76, p $$\le $$ ≤ 0.001 for the fluorogenic C$${_{12}}$$ 12 FDG method, and r = 0.72, p $$\le $$ ≤ 0.05 for the forward scatter method), and good fold difference ranges (1.120–4.436 for total autofluorescence mean and 1.082–6.362 for lipopigment autofluorescence mean) between early and senescent passage hMSCs. Our autofluroescence imaging and spectra decomposition platform offers additional benefit in label-free characterisation of senescent hMSC cells and could be further developed for adoption for future in situ cellular senescence evaluation by the cell manufacturers.

Download Full-text

Alloreactive Immune Response Associated to Human Mesenchymal Stromal Cells Treatment: A Systematic Review

Journal of Clinical Medicine ◽

10.3390/jcm10132991 ◽

2021 ◽

Vol 10 (13) ◽

pp. 2991

Author(s):

Raquel Sanabria-de la Torre ◽

María I. Quiñones-Vico ◽

Ana Fernández-González ◽

Manuel Sánchez-Díaz ◽

Trinidad Montero-Vílchez ◽

...

Keyword(s):

Systematic Review ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Immune Rejection ◽

Human Mesenchymal Stromal Cells ◽

Immunological Profile ◽

Tolerability Data ◽

Donor Specific Antibodies ◽

The Impact ◽

Hla Mismatches

The well-known immunomodulatory and regenerative properties of mesenchymal stromal cells (MSCs) are the reason why they are being used for the treatment of many diseases. Because they are considered hypoimmunogenic, MSCs treatments are performed without considering histocompatibility barriers and without anticipating possible immune rejections. However, recent preclinical studies describe the generation of alloantibodies and the immune rejection of MSCs. This has led to an increasing number of clinical trials evaluating the immunological profile of patients after treatment with MSCs. The objective of this systematic review was to evaluate the generation of donor specific antibodies (DSA) after allogeneic MSC (allo-MSC) therapy and the impact on safety or tolerability. Data from 555 patients were included in the systematic review, 356 were treated with allo-MSC and the rest were treated with placebo or control drugs. A mean of 11.51% of allo-MSC-treated patients developed DSA. Specifically, 14.95% of these patients developed DSA and 6.33% of them developed cPRA. Neither the production of DSA after treatment nor the presence of DSA at baseline (presensitization) were correlated with safety and/or tolerability of the treatment. The number of doses administrated and human leucocyte antigen (HLA) mismatches between donor and recipient did not affect the production of DSA. The safety of allo-MSC therapy has been proved in all the studies and the generation of alloantibodies might not have clinical relevance. However, there are very few studies in the area. More studies with adequate designs are needed to confirm these results.

Download Full-text