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. 2014 Mar 3;9(3):e89621.
doi: 10.1371/journal.pone.0089621. eCollection 2014.

Pentosan polysulfate decreases myocardial expression of the extracellular matrix enzyme ADAMTS4 and improves cardiac function in vivo in rats subjected to pressure overload by aortic banding

Maria Vistnes  1 , Jan Magnus Aronsen  2 , Ida G Lunde  3 , Ivar Sjaastad  1 , Cathrine R Carlson  1 , Geir Christensen  1
Affiliations

Pentosan polysulfate decreases myocardial expression of the extracellular matrix enzyme ADAMTS4 and improves cardiac function in vivo in rats subjected to pressure overload by aortic banding

Maria Vistnes et al. PLoS One. .

Abstract

Background: We hypothesized that cleavage of the extracellular matrix (ECM) proteoglycans versican and aggrecan by ADAMTS (a disintegrin and metalloprotease with thrombospondin motifs) proteases, which contributes to stress-induced ECM-reorganization in atherogenesis and osteoarthritis, also play a role in heart failure development.

Objectives: The primary objective was to identify alterations in expression of ADAMTS versicanases and aggrecanases during development of heart failure, while evaluation of the effects of in vivo modulation of relevant changes in ADAMTS activity constituted the secondary objective.

Methods: Myocardial levels of versican, aggrecan, and their ADAMTS cleaving proteases were examined in Wistar rats six weeks after aortic banding (AB), and versican and selected ADAMTS versicanases were further analyzed in neonatal cardiomyocytes (NCM) and cardiac fibroblasts (NFB) after stimulation by inflammatory mediators. Based on the initial findings, ADAMTS4 was selected the most promising therapeutic target. Thus, rats with AB were treated with pentosan polysulfate (PPS), a polysaccharide with known ADAMTS4-inhibitory properties, and effects on versican fragmentation, left ventricular function and geometry were evaluated.

Results: We discovered that myocardial mRNA and protein levels of ADAMTS1 and -4, and mRNA levels of versican, aggrecan, and ADAMTS8 increased after AB, and TNF-α and IL-1β synergistically increased mRNA of versican and ADAMTS4 in NCM and NFB and secretion of ADAMTS4 from NCM. Furthermore, PPS-treatment improved systolic function, demonstrated by an improved fractional shortening (vehicle 48±3% versus PPS 60±1%, p<0.01) after AB. Following PPS-treatment, we observed an ∼80% reduction in myocardial ADAMTS4 mRNA (p = 0.03), and ∼50% reduction in the extracellular amount of the p150 versican fragments (p = 0.05), suggesting reduced versicanase activity.

Conclusions: Our findings suggest that AB induces an increase in myocardial ADAMTS4 versicanase activity, and that PPS-treatment improved systolic function in the pressure-overloaded heart, holding promise as a novel therapeutic agent in heart failure.

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Conflict of interest statement

Competing Interests: The authors have read the journal's policy and have the following conflicts: A patent application has been submitted by the authors based on these results. We have filed a provisional patent application (use of enzyme inhibitors) with the UKIPO, in the name of University of Oslo, with the application number 1312311.2. The patent application does not alter our adherence to all the PLOS One policies on sharing data and materials.

Figures

Figure 1
Figure 1. Elevated mRNA levels of ADAMTS1, -4 and-8 in rats after aortic banding.
Myocardial mRNA expression of ADAMTS versicanases and aggrecanases in rats with preserved (HFpFS, n = 9) or reduced FS (HFrFS, n = 6) six weeks after AB, normalized to reference gene RPL4 and relative to sham (n = 9). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; FS, fractional shortening; ribosomal protein L4, RPL4.
Figure 2
Figure 2. Increased protein expression of ADAMTS1 and -4 after aortic banding.
Myocardial protein expression of ADAMTS1 (A) and -4 (B) in rats with preserved (HFpFS, n = 9) or reduced FS (HFrFS, n = 6) six weeks after aortic banding. Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; FS, fractional shortening.
Figure 3
Figure 3. Elevated mRNA levels of versican and aggrecan in rats after aortic banding.
Myocardial mRNA expression of versican and aggrecan in rats with preserved (HFpFS, n = 9) or reduced FS (HFrFS, n = 6) six weeks after AB, normalized to the reference gene RPL4 and relative to sham (n = 9). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; FS, fractional shortening; ribosomal protein L4, RPL4.
Figure 4
Figure 4. Myocardial TIMP-3, HAS and MT4-MMP mRNA levels in rats after aortic banding.
Myocardial mRNA expression of TIMP-3, HAS and MT4-MMP in rats with preserved (HFpFS, n = 9) or reduced FS (HFrFS, n = 6) six weeks after aortic banding, normalized to the reference gene ribosomal protein L4 (RPL4) and relative to sham (n = 9). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; FS, fractional shortening; TIMP, tissue inhibitor metalloprotease; HAS, hyaluronic acid synthase; MT, membrane-type.
Figure 5
Figure 5. ADAMTS4 and versican mRNA in cardiac fibroblasts and cardiomyocytes was induced by inflammatory mediators.
mRNA in NFB and NCM of ADAMTS1 (A,B), ADAMTS4 (C,D), and versican (E,F) after stimulation with LPS (n = 6), IL-1β (n = 7), TNF-α (n = 10 (NCM)/6 (NFB)), separate and combined (n = 7 (NCM)/4 (NFB)). Untreated cells served as control (n = 15 (NCM)/18 (NFB)). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05, compared to control. NFB, neonatal fibroblasts; NCM, neonatal cardiomyocytes; LPS, lipopolysaccharide; IL, interleukin; TNF, tumor necrosis factor.
Figure 6
Figure 6. Altered ADAMTS4 protein levels in cell medium from cardiac cells after stimulation with inflammatory mediators.
Protein levels of ADAMTS4 active forms (p68 and p53) in cell medium after stimulation of IL-1β alone (NFB n = 4, NCM n = 3) and in combination with TNF-α (NFB n = 3, NCM n = 4), relative to controls (n = 4). The bars represent mean levels and the error bars SEM. *p<0.05. NFB, neonatal fibroblasts; NCM, neonatal cardiomyocytes; LPS, lipopolysaccharide; IL, interleukin; TNF, tumor necrosis factor.
Figure 7
Figure 7. Decreased myocardial ADAMTS4 mRNA level after PPS-treatment.
Myocardial mRNA expression of ADAMTS4 in rats treated with PPS (n = 10) or vehicle (n = 7) six weeks after AB, and in sham-operated rats treated with PPS (n = 3) normalized to the reference gene RPL4 and relative to vehicle-treated sham (n = 3). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; PPS, pentosan polysulfate; HF, heart failure; RPL4, ribosomal protein L4.
Figure 8
Figure 8. Myocardial mRNA levels after PPS treatment in rats after aortic banding.
Myocardial mRNA expression of ADAMTS1, -5, -8 (A), versican, aggrecan (B), HAS-1, -2, TIMP-3, and MT4-MMP (C) in rats treated with PPS (n = 9 or 10) or vehicle (n = 7 or 9) six weeks after AB, and sham-operated rats treated with PPS (n = 3 or 4) normalized to the reference gene ribosomal protein L4 (RPL4) and relative to vehicle-treated sham (n = 3). The bars represent median levels and the error bars the 75th percentile. *p<0.05. HF, heart failure, HA, hyaluronic acid.
Figure 9
Figure 9. Versican p150 fragment level was reduced after PPS-treatment in rats with aortic banding.
Myocardial levels of p150 (A) and p70 (B) versican DPEEAE fragment in NaCl-soluble ECM of rats subjected to AB with or without PPS-treatment, shown as representative immunoblot and box-plots with median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; PPS, pentosan polysulfate; ECM, extracellular matrix; HF, heart failure.
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References

    1. Spinale FG, Janicki JS, Zile MR (2013) Membrane-Associated Matrix Proteolysis and Heart Failure. Circ Res 112: 195–208. - PMC - PubMed
    1. Dorman G, Kocsis-Szommer K, Spadoni C, Ferdinandy P (2007) MMP inhibitors in cardiac diseases: an update. Recent Pat Cardiovasc Drug Discov 2: 186–194. - PubMed
    1. Hudson MP, Armstrong PW, Ruzyllo W, Brum J, Cusmano L, et al. (2006) Effects of Selective Matrix Metalloproteinase Inhibitor (PG-116800) to Prevent Ventricular Remodeling After Myocardial Infarction: Results of the PREMIER (Prevention of Myocardial Infarction Early Remodeling) Trial. Journal of the American College of Cardiology 48: 15–20. - PubMed
    1. Waehre A, Halvorsen B, Yndestad A, Husberg C, Sjaastad I, et al. (2011) Lack of Chemokine Signaling through CXCR5 Causes Increased Mortality, Ventricular Dilatation and Deranged Matrix during Cardiac Pressure Overload. PLoS ONE 6: e18668 10.1371/journal.pone.0018668 - DOI - PMC - PubMed
    1. Strand ME, Herum KM, Rana ZA, Skrbic B, Askevold ET, et al. (2013) Innate immune signaling induces expression and shedding of the heparan sulfate proteoglycan syndecan-4 in cardiac fibroblasts and myocytes, affecting inflammation in the pressure-overloaded heart. FEBS J 280: 2228–2247. - PubMed

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Grants and funding

This work was supported by University of Oslo, Anders Jahre's Fund for the Promotion of Science (www.unifor.no/Fund.aspx?site=1&fund=290), the Kristian Gerhard Jebsen foundation (www.stiftkgj.no), the South-Eastern Norway Regional Health Authority (www.helse-sorost.no), the Research Council of Norway (www.forskningsradet.no), Norway, and the Simon Fougner Hartmanns Family Fund, Denmark. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.