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Osteopontin is indispensible for AP1-mediated angiotensin II-related miR-21 transcription during cardiac fibrosis.

Lorenzen JM, Schauerte C, Hübner A, Kölling M, Martino F, Scherf K, Batkai S, Zimmer K, Foinquinos A, Kaucsar T, Fiedler J, Kumarswamy R, Bang C, Hartmann D, Gupta SK, Kielstein J, Jungmann A, Katus HA, Weidemann F, Müller OJ, Haller H, Thum T - Eur. Heart J. (2015)

Bottom Line: OPN and miR-21 were significantly increased in cardiac biopsies of patients with myocardial fibrosis.This was associated with enhanced cardiac collagen content, myofibroblast activation, ERK-MAP kinase as well as AKT signalling pathway activation and a reduced expression of Phosphatase and Tensin Homologue (PTEN) as well as SMAD7 in WT but not OPN KO mice.In vitro, Ang II induced expression of miR-21 in WT cardiac fibroblasts, while miR-21 levels were unchanged in OPN KO fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular and Translational Therapeutic Strategies (IMTTS), Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany Department of Internal Medicine, Division of Nephrology and Hypertension, Hannover Medical School, Hannover, Germany lorenzen.johan@mh-hannover.de thum.thomas@mh-hannover.de.

No MeSH data available.


Related in: MedlinePlus

Ang II-induced fibrosis and miRNA expression in osteopontin wild type and knockout mice: Sirius red staining in paraffin-embedded sections of cardiac tissue of osteopontin wild type mice following vehicle-infusion (2 weeks, PBS, A) and Ang II infusion (B) as well as quantification of results (C) and osteopontin knockout mice following vehicle-infusion (D) and Ang II infusion (E) as well as quantification of results (F). Expression of Collagen I (G) and alpha smooth muscle actin (α-SMA, H) mRNA following Ang II or vehicle infusion. A global miRNA screen in hearts of osteopontin wild type and osteopontin knockout mice following Ang II infusion (I). MiR-21 expression in osteopontin wild type and knockout animals (J). n = 6 animals per group and analysis.
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EHV109F2: Ang II-induced fibrosis and miRNA expression in osteopontin wild type and knockout mice: Sirius red staining in paraffin-embedded sections of cardiac tissue of osteopontin wild type mice following vehicle-infusion (2 weeks, PBS, A) and Ang II infusion (B) as well as quantification of results (C) and osteopontin knockout mice following vehicle-infusion (D) and Ang II infusion (E) as well as quantification of results (F). Expression of Collagen I (G) and alpha smooth muscle actin (α-SMA, H) mRNA following Ang II or vehicle infusion. A global miRNA screen in hearts of osteopontin wild type and osteopontin knockout mice following Ang II infusion (I). MiR-21 expression in osteopontin wild type and knockout animals (J). n = 6 animals per group and analysis.

Mentions: In order to assess the in vivo relevance of OPN with respect to fibrosis development, we subjected OPN wild-type (WT) and OPN knockout (OPN KO) mice to chronic Ang II infusion via osmotic mini pumps for 2 weeks (Figure 2). In both groups, Ang II increased blood pressure (Supplementary material online, Table S2). Expectedly, we found that Ang II induced fibrosis in WT hearts as assessed by Sirius red staining (Figure 2A–C) and expression of the pro-fibrotic genes collagen I and α-SMA (Figure 2G and H). In contrast, in OPN KO mice, fibrosis development as well as fibrotic gene expression was strongly attenuated (Figure 2D–H). Similarly, Ang II upregulated various pro-fibrotic genes in cultured WT fibroblasts, including Collagen I alpha 2, Collagen III, matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), and α-SMA (Supplementary material online, Figure S3A–E), while there were no changes in OPN KO fibroblasts (Supplementary material online, Figure S3F–H). Ang II infusion in mice increased cardiomyocyte size in both groups (WT and OPN KO mice, Figure 3A–E), but only increased heart weight/body weight ratio in WT mice (Figure 3F), suggesting a prominent anti-fibrotic effect of OPN deficiency specifically in cardiac fibroblasts. Furthermore, Ang II induced pro-survival and pro-fibrotic signalling pathways in WT, but not in OPN KO hearts as assessed by AKT, SMAD3, and ERK phosphorylation (Figure 3G–J). Interestingly, in areas of increased collagen content (Sirius red staining), the expression of OPN was also increased, indicating a direct role in fibrosis development (Figure 3K and L). In WT fibroblasts, Ang II enhanced ERK phosphorylation as well as AKT phosphorylation (Supplementary material online, Figure S4A, B, F, and G). In addition, Ang II led to the nuclear exclusion of Foxo3a, which is associated with enhanced cellular survival (Supplementary material online, Figure S4D, J, and K). None of these events occurred in OPN KO fibroblasts (Supplementary material online, Figure S4C, E, H, I, L, and M).Figure 2


Osteopontin is indispensible for AP1-mediated angiotensin II-related miR-21 transcription during cardiac fibrosis.

Lorenzen JM, Schauerte C, Hübner A, Kölling M, Martino F, Scherf K, Batkai S, Zimmer K, Foinquinos A, Kaucsar T, Fiedler J, Kumarswamy R, Bang C, Hartmann D, Gupta SK, Kielstein J, Jungmann A, Katus HA, Weidemann F, Müller OJ, Haller H, Thum T - Eur. Heart J. (2015)

Ang II-induced fibrosis and miRNA expression in osteopontin wild type and knockout mice: Sirius red staining in paraffin-embedded sections of cardiac tissue of osteopontin wild type mice following vehicle-infusion (2 weeks, PBS, A) and Ang II infusion (B) as well as quantification of results (C) and osteopontin knockout mice following vehicle-infusion (D) and Ang II infusion (E) as well as quantification of results (F). Expression of Collagen I (G) and alpha smooth muscle actin (α-SMA, H) mRNA following Ang II or vehicle infusion. A global miRNA screen in hearts of osteopontin wild type and osteopontin knockout mice following Ang II infusion (I). MiR-21 expression in osteopontin wild type and knockout animals (J). n = 6 animals per group and analysis.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4543785&req=5

EHV109F2: Ang II-induced fibrosis and miRNA expression in osteopontin wild type and knockout mice: Sirius red staining in paraffin-embedded sections of cardiac tissue of osteopontin wild type mice following vehicle-infusion (2 weeks, PBS, A) and Ang II infusion (B) as well as quantification of results (C) and osteopontin knockout mice following vehicle-infusion (D) and Ang II infusion (E) as well as quantification of results (F). Expression of Collagen I (G) and alpha smooth muscle actin (α-SMA, H) mRNA following Ang II or vehicle infusion. A global miRNA screen in hearts of osteopontin wild type and osteopontin knockout mice following Ang II infusion (I). MiR-21 expression in osteopontin wild type and knockout animals (J). n = 6 animals per group and analysis.
Mentions: In order to assess the in vivo relevance of OPN with respect to fibrosis development, we subjected OPN wild-type (WT) and OPN knockout (OPN KO) mice to chronic Ang II infusion via osmotic mini pumps for 2 weeks (Figure 2). In both groups, Ang II increased blood pressure (Supplementary material online, Table S2). Expectedly, we found that Ang II induced fibrosis in WT hearts as assessed by Sirius red staining (Figure 2A–C) and expression of the pro-fibrotic genes collagen I and α-SMA (Figure 2G and H). In contrast, in OPN KO mice, fibrosis development as well as fibrotic gene expression was strongly attenuated (Figure 2D–H). Similarly, Ang II upregulated various pro-fibrotic genes in cultured WT fibroblasts, including Collagen I alpha 2, Collagen III, matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), and α-SMA (Supplementary material online, Figure S3A–E), while there were no changes in OPN KO fibroblasts (Supplementary material online, Figure S3F–H). Ang II infusion in mice increased cardiomyocyte size in both groups (WT and OPN KO mice, Figure 3A–E), but only increased heart weight/body weight ratio in WT mice (Figure 3F), suggesting a prominent anti-fibrotic effect of OPN deficiency specifically in cardiac fibroblasts. Furthermore, Ang II induced pro-survival and pro-fibrotic signalling pathways in WT, but not in OPN KO hearts as assessed by AKT, SMAD3, and ERK phosphorylation (Figure 3G–J). Interestingly, in areas of increased collagen content (Sirius red staining), the expression of OPN was also increased, indicating a direct role in fibrosis development (Figure 3K and L). In WT fibroblasts, Ang II enhanced ERK phosphorylation as well as AKT phosphorylation (Supplementary material online, Figure S4A, B, F, and G). In addition, Ang II led to the nuclear exclusion of Foxo3a, which is associated with enhanced cellular survival (Supplementary material online, Figure S4D, J, and K). None of these events occurred in OPN KO fibroblasts (Supplementary material online, Figure S4C, E, H, I, L, and M).Figure 2

Bottom Line: OPN and miR-21 were significantly increased in cardiac biopsies of patients with myocardial fibrosis.This was associated with enhanced cardiac collagen content, myofibroblast activation, ERK-MAP kinase as well as AKT signalling pathway activation and a reduced expression of Phosphatase and Tensin Homologue (PTEN) as well as SMAD7 in WT but not OPN KO mice.In vitro, Ang II induced expression of miR-21 in WT cardiac fibroblasts, while miR-21 levels were unchanged in OPN KO fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular and Translational Therapeutic Strategies (IMTTS), Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany Department of Internal Medicine, Division of Nephrology and Hypertension, Hannover Medical School, Hannover, Germany lorenzen.johan@mh-hannover.de thum.thomas@mh-hannover.de.

No MeSH data available.


Related in: MedlinePlus