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Analysis of the Secretome of Apoptotic Peripheral Blood Mononuclear Cells: Impact of Released Proteins and Exosomes for Tissue Regeneration.

Beer L, Zimmermann M, Mitterbauer A, Ellinger A, Gruber F, Narzt MS, Zellner M, Gyöngyösi M, Madlener S, Simader E, Gabriel C, Mildner M, Ankersmit HJ - Sci Rep (2015)

Bottom Line: Subsequently, in vitro assays showed that the exosome and protein fractions of irradiated and non-irradiated PBMC secretome were the major biological components that enhanced cell mobility; conversely, secreted lipids and microparticles had no effects.We found that the potency for preventing ventricular remodeling was similar with the GMP-compliant and experimentally-prepared PBMC secretomes.Our results indicate that irradiation modulates the release of proteins, lipid-mediators and extracellular vesicles from human PBMCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Medical University of Vienna, Austria.

ABSTRACT
We previously showed that, when peripheral blood mononuclear cells (PBMCs) were stressed with ionizing radiation, they released paracrine factors that showed regenerative capacity in vitro and in vivo. This study aimed to characterize the secretome of PBMCs and to investigate its biologically active components in vitro and vivo. Bioinformatics analysis revealed that irradiated PBMCs differentially expressed genes that encoded secreted proteins. These genes were primarily involved in (a) pro-angiogenic and regenerative pathways and (b) the generation of oxidized phospholipids with known pro-angiogenic and inflammation-modulating properties. Subsequently, in vitro assays showed that the exosome and protein fractions of irradiated and non-irradiated PBMC secretome were the major biological components that enhanced cell mobility; conversely, secreted lipids and microparticles had no effects. We tested a viral-cleared PBMC secretome, prepared according to good manufacturing practice (GMP), in a porcine model of closed chest, acute myocardial infarction. We found that the potency for preventing ventricular remodeling was similar with the GMP-compliant and experimentally-prepared PBMC secretomes. Our results indicate that irradiation modulates the release of proteins, lipid-mediators and extracellular vesicles from human PBMCs. In addition our findings implicate the use of secretome fractions as valuable material for the development of cell-free therapies in regenerative medicine.

No MeSH data available.


Related in: MedlinePlus

Analysis of changes in PBMC transcriptome.PBMCs were either irradiated or not irradiated ex vivo, before culturing for 2, 4, or 20 h. At the indicated time points, total RNA was isolated, and microarray analyses were performed to evaluate gene expression. (Left) Venn diagrams show the numbers of upregulated transcripts with expression changes >2-fold above expression after cell separation for (a) non-irradiated and (c) irradiated PBMCs. Each circle depicts the genes detected at the indicated time point; overlapping sections indicate the number of genes that were upregulated at multiple time points. (Right) The heatmaps show that a significant number of genes were upregulated in (b) non-irradiated and (d) irradiated PBMCs with a strong time dependence, and expression was most prominent at 20 h after cultivation; green = downregulated; red = upregulated in cultured samples; n = 4 for each time point.
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f2: Analysis of changes in PBMC transcriptome.PBMCs were either irradiated or not irradiated ex vivo, before culturing for 2, 4, or 20 h. At the indicated time points, total RNA was isolated, and microarray analyses were performed to evaluate gene expression. (Left) Venn diagrams show the numbers of upregulated transcripts with expression changes >2-fold above expression after cell separation for (a) non-irradiated and (c) irradiated PBMCs. Each circle depicts the genes detected at the indicated time point; overlapping sections indicate the number of genes that were upregulated at multiple time points. (Right) The heatmaps show that a significant number of genes were upregulated in (b) non-irradiated and (d) irradiated PBMCs with a strong time dependence, and expression was most prominent at 20 h after cultivation; green = downregulated; red = upregulated in cultured samples; n = 4 for each time point.

Mentions: We first identified transcripts that were upregulated during the cultivation period in either non-irradiated (Fig. 2a) or irradiated PBMCs (Fig. 2c). We only analyzed transcripts with a FC ≥ 2 compared to baseline values. The heatmaps displayed 525 and 1099 genes that were upregulated in at least two of the three time points in non-irradiated (Fig. 2b) and irradiated PBMCs (Fig. 2d), respectively. The bioinformatics analysis identified 167 transcripts that encoded actively secreted proteins in non-irradiated PBMCs and 213 that encoded secreted proteins in irradiated PBMCs (supplementary Table S1).


Analysis of the Secretome of Apoptotic Peripheral Blood Mononuclear Cells: Impact of Released Proteins and Exosomes for Tissue Regeneration.

Beer L, Zimmermann M, Mitterbauer A, Ellinger A, Gruber F, Narzt MS, Zellner M, Gyöngyösi M, Madlener S, Simader E, Gabriel C, Mildner M, Ankersmit HJ - Sci Rep (2015)

Analysis of changes in PBMC transcriptome.PBMCs were either irradiated or not irradiated ex vivo, before culturing for 2, 4, or 20 h. At the indicated time points, total RNA was isolated, and microarray analyses were performed to evaluate gene expression. (Left) Venn diagrams show the numbers of upregulated transcripts with expression changes >2-fold above expression after cell separation for (a) non-irradiated and (c) irradiated PBMCs. Each circle depicts the genes detected at the indicated time point; overlapping sections indicate the number of genes that were upregulated at multiple time points. (Right) The heatmaps show that a significant number of genes were upregulated in (b) non-irradiated and (d) irradiated PBMCs with a strong time dependence, and expression was most prominent at 20 h after cultivation; green = downregulated; red = upregulated in cultured samples; n = 4 for each time point.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4645175&req=5

f2: Analysis of changes in PBMC transcriptome.PBMCs were either irradiated or not irradiated ex vivo, before culturing for 2, 4, or 20 h. At the indicated time points, total RNA was isolated, and microarray analyses were performed to evaluate gene expression. (Left) Venn diagrams show the numbers of upregulated transcripts with expression changes >2-fold above expression after cell separation for (a) non-irradiated and (c) irradiated PBMCs. Each circle depicts the genes detected at the indicated time point; overlapping sections indicate the number of genes that were upregulated at multiple time points. (Right) The heatmaps show that a significant number of genes were upregulated in (b) non-irradiated and (d) irradiated PBMCs with a strong time dependence, and expression was most prominent at 20 h after cultivation; green = downregulated; red = upregulated in cultured samples; n = 4 for each time point.
Mentions: We first identified transcripts that were upregulated during the cultivation period in either non-irradiated (Fig. 2a) or irradiated PBMCs (Fig. 2c). We only analyzed transcripts with a FC ≥ 2 compared to baseline values. The heatmaps displayed 525 and 1099 genes that were upregulated in at least two of the three time points in non-irradiated (Fig. 2b) and irradiated PBMCs (Fig. 2d), respectively. The bioinformatics analysis identified 167 transcripts that encoded actively secreted proteins in non-irradiated PBMCs and 213 that encoded secreted proteins in irradiated PBMCs (supplementary Table S1).

Bottom Line: Subsequently, in vitro assays showed that the exosome and protein fractions of irradiated and non-irradiated PBMC secretome were the major biological components that enhanced cell mobility; conversely, secreted lipids and microparticles had no effects.We found that the potency for preventing ventricular remodeling was similar with the GMP-compliant and experimentally-prepared PBMC secretomes.Our results indicate that irradiation modulates the release of proteins, lipid-mediators and extracellular vesicles from human PBMCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Medical University of Vienna, Austria.

ABSTRACT
We previously showed that, when peripheral blood mononuclear cells (PBMCs) were stressed with ionizing radiation, they released paracrine factors that showed regenerative capacity in vitro and in vivo. This study aimed to characterize the secretome of PBMCs and to investigate its biologically active components in vitro and vivo. Bioinformatics analysis revealed that irradiated PBMCs differentially expressed genes that encoded secreted proteins. These genes were primarily involved in (a) pro-angiogenic and regenerative pathways and (b) the generation of oxidized phospholipids with known pro-angiogenic and inflammation-modulating properties. Subsequently, in vitro assays showed that the exosome and protein fractions of irradiated and non-irradiated PBMC secretome were the major biological components that enhanced cell mobility; conversely, secreted lipids and microparticles had no effects. We tested a viral-cleared PBMC secretome, prepared according to good manufacturing practice (GMP), in a porcine model of closed chest, acute myocardial infarction. We found that the potency for preventing ventricular remodeling was similar with the GMP-compliant and experimentally-prepared PBMC secretomes. Our results indicate that irradiation modulates the release of proteins, lipid-mediators and extracellular vesicles from human PBMCs. In addition our findings implicate the use of secretome fractions as valuable material for the development of cell-free therapies in regenerative medicine.

No MeSH data available.


Related in: MedlinePlus