Discovery of Thymosin Beta-4 as a Human Exerkine and Growth Factor

2021 ◽  
Author(s):  
Alba Gonzalez-Franquesa ◽  
Ben Stocks ◽  
Melissa L. Borg ◽  
Michael Kuefner ◽  
Emilie Dalbram ◽  
...  
Keyword(s):  
Growth Factor ◽  
Potential Role ◽  
C2c12 Myotubes ◽  
Osteoblast Proliferation ◽  
Endocrine Organ ◽  
Thymosin Beta 4 ◽  
Diet Induced Obesity ◽  
The Media ◽  
Exercise Induced

Skeletal muscle is an endocrine organ secreting exercise-induced factors (exerkines), which play a pivotal role in inter-organ crosstalk. Using mass spectrometry (MS)-based proteomics, we characterized the secretome and identified thymosin beta-4 (TMSB4X) as the most upregulated secreted protein in the media of contracting C2C12 myotubes. TMSB4X was also acutely increased in plasma of exercising humans irrespective of the insulin resistance condition or exercise mode. Treatment of mice with TMSB4X did not ameliorate the metabolic disruptions associated with diet induced-obesity, nor did it enhance muscle regeneration in vivo. However, TMSB4X increased osteoblast proliferation and neurite outgrowth, consistent with its WADA-classification as a prohibited growth factor. Therefore, we report TMSB4X as a human exerkine with a potential role in cellular crosstalk.

Absence of transforming growth factor beta 1 in murine platelets reduces neointima formation without affecting arterial thrombosis

2017 ◽  
Vol 117 (09) ◽  
pp. 1782-1797 ◽  
Author(s):  
Eva Schütz ◽  
Magdalena L. Bochenek ◽  
Dennis R. Riehl ◽  
Markus Bosmann ◽  
Thomas Münzel ◽  
...  
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Arterial Injury ◽  
Neointima Formation ◽  
Proliferating Cells ◽  
The Media

SummaryPlatelet degranulation at the site of vascular injury prevents bleeding and may affect the chronic vascular wound healing response. Transforming Growth Factor (TGF)-β1 is a major component of platelet α-granules known to accumulating in thrombi. It was our aim to determine the role of TGFβ1 released from activated platelets for neointima formation following arterial injury and thrombosis. Mice with platelet-specific deletion of TGFβ1 (Plt.TGFβ-KO) underwent carotid artery injury. Immunoassays confirmed the absence of active TGFβ1 in platelet releasates and plasma of Plt.TGFβ-KO mice. Whole blood analyses revealed similar haematological parameters, and tail cut assays excluded major bleeding defects. Platelet aggregation and the acute thrombotic response to injury in vivo did not differ between Plt.TGFβ-KO and Plt.TGFβ-WT mice. Morphometric analysis revealed that absence of TGFβ1 in platelets resulted in a significant reduction of neointima formation with lower neointima area, intima-to-media ratio, and lumen stenosis. On the other hand, the media area was enlarged in mice lacking TGFβ1 in platelets and contained increased amounts of proteases involved in latent TGFβ activation, including MMP2, MMP9 and thrombin. Significantly increased numbers of proliferating cells and cells expressing the mesenchymal markers platelet-derived growth factor receptor-β or fibroblast-specific protein-1, and the macrophage antigen F4/80, were observed in the media of Plt.TGFβ-KO mice, whereas the medial smooth muscle-actin-immuno-positive area and collagen content did not differ between genotypes. Our findings support an essential role for platelet-derived TGFβ1 for the vascular remodelling response to arterial injury, apparently independent from the role of platelets in thrombosis or haemostasis.Supplementary Material to this article is available online at www.thrombosis-online.com.

scholarly journals Sustained Release of Transforming Growth Factor-β1 from Platelet-Rich Chondroitin Sulfate Glycosaminoglycan Gels

2017 ◽  
Vol 31 (05) ◽  
pp. 410-415 ◽  
Author(s):  
Kate Birdwhistell ◽  
Lohitash Karumbaiah ◽  
Samuel Franklin
Keyword(s):  
Growth Factor ◽  
Chondroitin Sulfate ◽  
Cartilage Repair ◽  
Transforming Growth Factor ◽  
Platelet Rich Plasma ◽  
Enzyme Linked Immunosorbent Assay ◽  
The Media ◽  
Tgf Β1

AbstractActivated platelet-rich plasma (PRP), also referred to as platelet-rich fibrin (PRF), has been used to augment numerous techniques of cartilage repair in the knee but does not always result in superior quality of repair tissue. One possible reason that PRF does not consistently result in excellent cartilage regeneration is the transiency of growth factor provision with PRF. The objective of this study was to compare the release of transforming growth factor (TGF)-β1 from PRF and from PRP combined with a novel chondroitin sulfate glycosaminoglycan (CS-GAG) gel. PRP was prepared from nine healthy dogs and split into two aliquots: one activated with bovine thrombin and calcium chloride (CaCl2) to form PRF and the other aliquot was used to rehydrate a lyophilized CS-GAG gel. Both PRF and the CS-GAG gels were incubated in media for 13 days and media were collected, stored, and replaced every 48 hours and the concentration of TGF-β1 quantified in the media using an enzyme-linked immunosorbent assay. Concentrations of TGF-β1 in the media were up to three times greater with the CS-GAG gels and were significantly (p < 0.05) greater than with PRF on days 3, 5, 7, 9, and 13. Furthermore, TGF-β1 elution was still substantial at day 13 with the use of the CS-GAG gels. Additional in vitro work is warranted to characterize TGF-β1 elution from this CS-GAG gel with human PRP and to determine whether the use of these CS-GAG gels can augment cartilage repair in vivo.

scholarly journals Modulation of collagen synthesis by a growth factor and by the extracellular matrix: comparison of cellular response to two different stimuli.

1983 ◽  
Vol 97 (3) ◽  
pp. 803-809 ◽  
Author(s):  
S C Tseng ◽  
N Savion ◽  
D Gospodarowicz ◽  
R Stern
Keyword(s):  
Extracellular Matrix ◽  
Growth Factor ◽  
Cellular Response ◽  
Collagen Synthesis ◽  
Basal Surface ◽  
Matrix Deposition ◽  
The Matrix ◽  
The Media ◽  
In Cells

Cultured bovine corneal endothelial cells can be grown in three ways: on plastic, on plastic with fibroblast growth factor present in the media, and on their own preformed extracellular matrix. On plastic alone, cells grow in a disorderly fashion and secrete matrix on all cell surfaces. Cells grown on plastic with growth factor or on a matrix, at confluence, have matrix deposition only on the basal surface of the cells and an orderly contact-inhibited pattern of growth. This correlates with the polarity they demonstrate histologically. This cell-matrix pattern resembles the pattern observed in vivo. Both the soluble growth factor and the extracellular matrix are able to modulate the pattern of collagen synthesis and deposition by cells, but they do so in two entirely different ways. In cells grown on the extracellular matrix, total collagen synthesis is lower but more efficient. Collagen is deposited primarily into the cell layer even at the early sparse stage of culture. In cells grown on plastic with growth factor in the media, collagen is initially secreted into the media and does not become incorporated into the matrix. The deposition of collagen on the basal surface of cell occurs only late in the culture, and is achieved by increments in a stepwise manner. The in vivo-like pattern is not manifest until confluence has been reached. Thus, the extracellular matrix functions not only as a structural support, but is also instructional to the cells plated on it. In this case, the matrix regulates the level of collagen synthesis in the cells and modulates the pattern of collagen deposition. Soluble growth factors may act in part by enhancing a cell's ability to elaborate an appropriate matrix pattern necessary for the cell's own growth and accurate function.

scholarly journals Influence of statins on distinct circulating microRNAs during prolonged aerobic exercise

2016 ◽  
Vol 120 (6) ◽  
pp. 711-720 ◽  
Author(s):  
Pil-Ki Min ◽  
Joseph Park ◽  
Stephanie Isaacs ◽  
Beth A. Taylor ◽  
Paul D. Thompson ◽  
...  
Keyword(s):  
Muscle Contraction ◽  
Muscle Injury ◽  
C2c12 Myotubes ◽  
Circulating Microrna ◽  
Extracellular Release ◽  
Boston Marathon ◽  
Exercise Induced ◽  
Statin Use

Statins exacerbate exercise-induced skeletal muscle injury. Muscle-specific microRNAs (myomiRs) increase in plasma after prolonged exercise, but the patterns of myomiRs release after statin-associated muscle injury have not been examined. We examined the relationships between statin exposure, in vitro and in vivo muscle contraction, and expression of candidate circulating myomiRs. We measured plasma levels of myomiRs, circulating microRNA-1 (c-miR-1), c-miR-133a, c-miR-206, and c-miR-499-5p from 28 statin-using and 28 nonstatin-using runners before (PRE), immediately after (FINISH), and 24 h after they ran a 42-km footrace (the 2011 Boston marathon) (POST-24). To examine these cellular-regulation myomiRs, we used contracting mouse C2C12 myotubes in culture with and without statin exposure to compare intracellular and extracellular expression of these molecules. In marathoners, c-miR-1, c-miR-133a, and c-miR-206 increased at FINISH, returned to baseline at POST-24, and were unaffected by statin use. In contrast, c-miR-499-5p was unchanged at FINISH but increased at POST-24 among statin users compared with PRE and runners who did not take statins. In cultured C2C12 myotubes, extracellular c-miR-1, c-miR-133a, and c-miR-206 were significantly increased by muscle contraction regardless of statin use. In contrast, extracellular miR-499-5p was unaffected by either isolated statin exposure or isolated carbachol exposure but it was increased when muscle contraction was combined with statin exposure. In summary, we found that statin-potentiated muscle injury during exercise is accompanied by augmented extracellular release of miR-499-5p. Thus c-miR-499-5p may serve as a biomarker of statin-potentiated muscle damage.

scholarly journals Mutation and Analysis of Dan, the Founding Member of the Dan Family of Transforming Growth Factor β Antagonists

2001 ◽  
Vol 21 (2) ◽  
pp. 636-643 ◽  
Author(s):  
Marc S. Dionne ◽  
William C. Skarnes ◽  
Richard M. Harland
Keyword(s):  
Growth Factor ◽  
Potential Role ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Axonal Outgrowth ◽  
Frog Embryo ◽  
Founding Member ◽  
Evolutionarily Conserved ◽  
Developing Neurons

ABSTRACT The Dan family of transforming growth factor β antagonists is a large, evolutionarily conserved family of proteins. Little is known about either the specificity of these antagonists or the biological roles of these proteins. We have characterized Dan, the founding member of this family, with regard to both its biochemical specificity and its biological roles. Although DAN is not an efficient antagonist of BMP-2/4 class signals, we found that DAN was able to interact with GDF-5 in a frog embryo assay, suggesting that DAN may regulate signaling by the GDF-5/6/7 class of BMPs in vivo. Intriguingly, in developing neurons, Dan mRNA was localized to axons, suggesting a potential role for the DAN protein in axonal outgrowth or guidance. Mice lacking Dan activity were generated by gene targeting and displayed subtle, background-dependent defects.

scholarly journals Growth Factor-Dependent Activation of a MYC-Induced Latent AML Program in Human Hematopoietic Cells

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 2533-2533
Author(s):  
Elizabeth Bulaeva ◽  
Davide Pellacani ◽  
Naoto Nakamichi ◽  
Philip Beer ◽  
Colin Hammond ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Potential Role ◽  
Lymphoid Cells ◽  
Current Evidence ◽  
In Vivo Model ◽  
Hematopoietic Stem ◽  
Gm Csf

Background: Current evidence suggests that genetic and epigenetic abnormalities drive the development of human Acute Myeloid Leukemias (AMLs). However, whether these are sufficient to establish a permanent, self-sustaining AML population, and the potential role of shared perturbed downstream pathways is unknown. We hypothesized that a modest upregulated expression of MYC might play such a role given its commonly increased expression in many AML patients' cells. To test this hypothesis, we assessed the dynamics and types of cells produced in sublethally irradiated NOD-Rag1-/--IL2Rγc-/-(NRG) mice transgenically producing human IL3, GM-CSF and SCF (NRG-3GS mice) following their transplantation with freshly isolated subsets of normal CD34+ cord blood (CB) cells that were first lentivirally transduced with a human MYC cDNA. Results: FACS and Western blot analyses indicated this produced a 2 to 5-fold increase in MYC mRNA and protein levels in MYC-transduced CD34+ CB cells, and 21/22 NRG-3GS mice injected with ≥6,500 of these cells developed a fatal human AML population within 7 weeks. Histological analysis of their bone marrow and spleen cells showed both contained a prominent human CD123+CD33+CD15±CD34-CD14-CD19-CD3- blast population. Additional limiting dilution transplants showed that both the CD34+CD38- cells (enriched for hematopoietic stem cells) and the more differentiated CD34+ GMPs were similarly highly susceptible (at frequencies of 1/14 and 1/46, respectively) and, in both cases, generated progeny that could initiate serially transplantable leukemias with the same phenotypic and transcriptomic features. Comparison to normal CB cells indicated these most closely resembled GMPs, and comparison to pediatric AML patient samples indicated a similarity to myelomonocytic leukemias with enhanced MYC expression. Interestingly, 14 sublethally irradiated NRG mice (the parental strain not producing human 3GS) transplanted with matched aliquots of CD34+ MYC-transduced cells regenerated a normal spectrum of CD19+ lymphoid cells, CD14+ and CD15+ GM cells and readily detectable CD34+ cells for up to 32 weeks of follow-up with no evidence of leukemogenesis. However, transfer of these regenerated human cells into secondary NRG-3GS mice, even after this extended period, enabled their rapid production of a lethal human AML in all 5 mice tested. In contrast, matched aliquots transplanted into 5 NRG recipients produced declining grafts of normal cells. This finding was then exploited to determine which growth factors were responsible for activating the AML program by transplanting NRG mice with CD34+ CB cells transduced with MYC and just a single growth factor, or all 3 as a positive control. In this set of experiments, a lethal human AML was obtained when MYC was paired with human IL3 or GM-CSF (or all 3 together), but not with SCF (or no growth factors). Conclusion: We report here a new in vivo model of MYC-induced human myeloid leukemogenesis that produces a serially transplantable AML closely resembling human pediatric myelomonocytic leukemias with elevated MYC expression. The rapidity, consistency, and high frequency of this transformation process obtained by transducing late granulopoietic as well as early types of normal human CD34+ progenitor cells makes this system highly attractive for future mechanistic and therapeutic testing experiments. The discovery that MYC deregulation alone generates a stable "latent program" that can be rapidly activated by exposure to exogenous growth factors typical of inflammatory states also raises intriguing questions about the potential role of such events in the genesis of AML populations that arise in patients. Disclosures Beer: Karus therapeutics Ltd.: Employment.

Abstract 15255: Statins Augment Circulating MicroRNA-499-5p Release With Muscle Contraction and Aerobic Exercise

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Pil-Ki Min ◽  
Joseph Park ◽  
Stephanie Isaacs ◽  
Beth A Taylor ◽  
Paul D Thompson ◽  
...  
Keyword(s):  
Muscle Contraction ◽  
Muscle Injury ◽  
Fold Change ◽  
C2c12 Myotubes ◽  
Circulating Microrna ◽  
Cultured Myotubes ◽  
Exercise Induced ◽  
Statin Use

Background: Statins exacerbate exercise-induced skeletal muscle injury. Muscle-specific microRNAs (myomiRs) increase in plasma after prolonged exercise, but the patterns of myomiRs release after statin-associated muscle injury have not been examined. Objectives: We examined the relationships among statin exposure, in vitro and in vivo muscle contraction, and candidate circulating myomiRs expression. Methods: We measured plasma levels of the myomiRs, circulating microRNA-1 (c-miR-1), c-miR-133a, c-miR-206 and c-miR-499-5p, from 28 statin-using and 28 non-statin using runners before (PRE), immediately after (FINISH), and 24 hours after a 42 km footrace (POST-24). We subsequently used contracting mouse C2C12 cultured myotubes with and without statin exposure to compare intracellular and extracellular expression of these molecules. Fold-changes of microRNAs are presented as median [interquartile range]. Results: In marathoners, c-miR-1, c-miR-133a, and c-miR-206 increased at FINISH, returned to baseline at POST-24, and were unaffected by statin use. In contrast, c-miR-499-5p was unchanged at FINISH in both groups, but c-miR-499-5p increased in statin users at the POST-24 time point compared to PRE [2.9 (1.3, 8.6) vs. 1.0 fold change, p <0.001] and to non-statin using runners [2.9 (1.3, 8.6) vs. 1.4 (0.9, 3.2) fold change, p < 0.05]. In cultured C2C12 myotubes, intracellular levels of candidate myomiRs remained stable except for modest declines of miR-1 and miR-206 with isolated myotube contraction (carbachol exposure) or simultaneous statin and myotube contraction. Extracellular miR-1, 133a, and 206 increased with contraction regardless of statin use. In contrast, extracellular miR-499-5p was unaffected by either isolated statin exposure or isolated contraction but increased with contraction + statin [4.8 (1.9, 8.1) vs. 1.0 (0.7, 1.5) fold change, p < 0.05 vs. control]. Conclusions: Statin-potentiated muscle injury during exercise is accompanied by augmented extracellular release of miR-499-5p. c-miR-499-5p may serve as a biomarker of statin-potentiated muscle damage.

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