Recovery of Myocardial Infarction via Unique Modulation of the Cardiac Microenvironment
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1 2016 춘계심혈관통합학술대회, 경주 Recovery of Myocardial Infarction via Unique Modulation of the Cardiac Microenvironment Youngkeun Ahn, MD, PhD Department of Cardiology, Cardiovascular Center Chonnam National University Hospital
2 Main Events in Cardiac Injury Cardiomyocytes Loss Fibrosis Delayed Angiogenesis Remodeling & Dysfunction Uncontrolled Inflammation
3 Current Strategies for Cardiac Repair Well-Being Life Style Medical Check-Up Prevention PCI Statins Conventional Therapy (Attenuation) ACEi -Blockers Reprogramming Cocktail ipsc-cm Fb-CM Regeneration Primed MSC ESC-CM MSC ipsc CSC
4 Cardiac Regeneration by New CM 2016 Circ Res
5 Cardiac Regeneration by Nrg1 cardiomyocyte 2015 Nat Cell Biol, 2015 elife, 2015 Sci Trans Med, 2015 Nature
6 Cardiac Regeneration by FSTL1 2015, Nature, 2016 NEJM
7 Myocytes and Nonmyocytes in the Myocardium By Cell Vol. By Cell Mass By Cell No. Mostly Fb 20~33 90~95 Fb ~10 25~35 67~80 ~90 65~75 Myocytes Nonmyocytes
8 Introduction of BIO Tyrian purple dye Kim YS, PhD Jeong HY, MS 2 Z,3 E-6-Bromoindirubin-3 -oxime No Further Study 2006 Chem Biol
9 Proliferating CM (fold) Proliferating cfb (fold) Effect of BIO on Proliferation of CM and cfb * * BIO ( M), 5d Tx BIO ( M), 5d Tx 0 * *
10 Effect of BIO on Proliferation of CM and cfb Proliferating cells Non-proliferating cells CM (% fold) BIO 2 days cfb (% fold) * DAPI (blue) BrdU (green) TnI (red) BIO 5 days CM (% fold) * 5 µm BIO for 2 days 5 µm BIO for 5 days BIO 5 days cfb (% fold) BrdU(+)cTnI(+) Proliferating CM *
11 Cardiac Fibroblasts Fibroblasts Proliferation Migration TGF- Inflammatory mediators ECM Pathological Fibrosis
12 20 mm LiCl, BIO 3 days Tx Effect of BIO on Pro-fibrotic cfb
13 BIO 5 days Tx Effect of BIO on Akt and p27 in cfb
14 MI tissue infiltrate Cardiac Macrophages 3 7 days post MI IL-1 TNF- IL-12 Arg1 IL-10 CD206 NO M1 M2 Inflammatory phase Reparative phase Arter Thromb Vasc Biol 2009, J Am Coll Cardiol 2012, Circ Res 2014
15 inos NOHA NO + L-Citrulline Cytotoxicity Inflammation L-Arginine Arginase-1 L-Ornithine + urea OAT ODC Putrescine P5C L-Proline Spermidine Collagen Spermine Tissue Regeneration Cell Proliferation Anti-inflammation
16 Cardiac Recovery by Macrophage Modulation J Cell Mol Med 2014, Sci Rep 2015
17 Anti-inflammatory Effect of BIO on RAW264.7 cells LPS (100 ng/ml) treatment for 24 hours 20 mm LiCl
18 Fibroblasts and Macrophages 2013 J Mol Cell Cardiol
19 IL-10 mrna (fold) Arginase activity (fold) Effect of BIO on Crosstalk between cfb and (1) Macrophages * * LPS BIO LiCl Co-culture with cfb Macrophages * * * - LPS LPS LPS 5 µm BIO or 10 ng/ml IL-4 20 mm LiCl Co-culture of cfb with THP-1 MO for 72 h induced MФ differentiation - - IL4 BIO LiCl BIO LiCl
20 Arginase activity (fold) IL-10 mrna (fold) Effect of BIO on Crosstalk between cfb and (2) * * TGF - LPS LPS LPS IL4 - - BIO LiCl 50 ng/ml TGF-β * * * - LPS LPS LPS - - IL4 BIO LiCl BIO LiCl
21 Effect of BIO on Activated Macrophages inos - LPS - - BIO GAPDH TAK1 4hr 24hr Control LPS LPS+BIO LPS LPS+BIO IKKα IKKβ TBK1 γ(nemo ) TANK TBK1IKKε IKKγ(NEMO) GAPDH Raw cell 264.7, BIO 5μM
22 Effect of BIO on IKK in Macrophages Nucleus Cytosol Control LPS LPS/BIO Control LPS LPS/BIO pikkε Laminin B GAPDH
23 Survival rate (%) BrdU(+) CM Ventricle area (pixels) Cardiac Regeneration in Zebra Fish HF Model * - AA AA AA * * - AA AA AA - - BIO LiCl - - BIO LiCl - - BIO 72 hpf Tg(cmlc2:GFP) transgenic zebrafish larvae, 2.5 µm aristolochic acid for 3 h * - AA AA
24 Therapeutic Effect of BIO on MI Model 0.2 mg/kg BIO treatment
25
26 Anti-inflammatory in BIO/MI Group MI+Veh: CD68(+)Arg1(+) MI+BIO: CD68(+)Arg1(+) DAPI CD68 DAPI CD68 Arg1 Merge Arg1 Merge
27 Anti-inflammatoroy M
28 Microarray Analysis of BIO-treated CM D mrna (fold) E mrna (fold) 0 BIO (hr) BIO (hr)
29 Summary: Pleiotropic Effects on Cardiac Environment Cells BIO enhances the proliferation of CM and reduces the proliferation of Fb. BIO produces anti-fibrotic effects in Fb. BIO promotes monocyte differentiation into anti-inflammatory M. These beneficial effects of BIO can be observed in a zebrafish HF model and a rat MI model. This study supports the further development of BIO as a drug to target the cardiac microenvironment and improve remodeling after MI. F/E of BIO derivatives
30
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