CRTH2 promotes endoplasmic reticulum stress‐induced cardiomyocyte apoptosis through m‐calpain

• DOI: 10.15252/emmm.201708237
• Publication Date: 2018-01-16T01:10:19Z
• Authors: Shengkai Zuo; Deping Kong; Chenyao Wang; Jiao Liu; Yuanyang Wang; Qiangyou Wan; Shuai Yan; Jian Zhang; Juan Tang; Qianqian Zhang; Luheng Lyu; Xin Li; Zhixin Shan; Li Qian; Yujun Shen; Ying Yu
• Keywords: Christian ministry; Biological sciences

Research Article15 January 2018Open Access Source DataTransparent process CRTH2 promotes endoplasmic reticulum stress-induced cardiomyocyte apoptosis through m-calpain Shengkai Zuo Department of Pharmacology, Key Laboratory Immune Microenvironment and Disease (Ministry Education), School Basic Medical Sciences, Tianjin University, Tianjin, China Food Safety Research, Institute for Nutritional Shanghai Institutes Biological Chinese Academy Shanghai, Search more papers by this author Deping Kong Chenyao Wang orcid.org/0000-0002-0957-1956 Jiao Liu Yuanyang Qiangyou Wan Shuai Yan Jian Zhang Juan Tang Qianqian Luheng Lyu Biology, University Miami College Arts Science, Miami, FL, USA Xin Li Zhixin Shan Guangdong General Hospital, Cardiovascular Institute, Guangzhou, Guangdong, Qian McAllister Heart North Carolina at Chapel Hill, NC, Yujun Shen Corresponding Author [email protected] orcid.org/0000-0002-9266-9064 Ying Yu orcid.org/0000-0002-6476-1752 Information Zuo1,2, Kong1, Wang2, Liu2, Wang1, Wan2, Yan2, Zhang1, Tang2, Zhang2, Lyu2,3, Li1, Shan4, Qian5, *,1 *,1,2 1Department 2Key 3Department 4Medical 5McAllister *Corresponding author. Tel/Fax: [phone]; E-mail: EMBO Mol Med (2018)10:e8237https://doi.org/10.15252/emmm.201708237 PDFDownload PDF article text main figures. Peer ReviewDownload a summary the editorial decision including letters, reviewer comments responses to feedback. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Apoptotic death cardiac myocytes is associated with ischemic heart disease chemotherapy-induced cardiomyopathy. Chemoattractant receptor-homologous molecule expressed on T helper type 2 cells (CRTH2) highly in heart. However, its specific role cardiomyopathy not fully understood. Here, we demonstrated that disruption markedly improved recovery mice postmyocardial infarction doxorubicin challenge suppressing apoptosis. Mechanistically, activation specifically facilitated (ER) via caspase-12-dependent pathway. Blockage prevented CRTH2-mediated under ER stress caspase-12 activity. was coupled Gαq elicit intracellular Ca2+ flux activated m-calpain/caspase-12 cascade cardiomyocytes. Knockdown caspase-4, an alternative humans, alleviated CRHT2 activation-induced human response anoxia. Our findings revealed unexpected promoting apoptosis, suggesting inhibition has therapeutic potential Synopsis T-helper type-2 (CRTH2), which mediates chemoattractant effect prostaglandin (PG) D2 leukocytes, shown here promote PGD2/CRTH2 signaling cardiomyocytes upon such as anoxia (DOX) challenge. Gαq/m-calpain/caspase-12 confers cardioprotection against myocardial infaction or DOX treatment reducing Introduction diseases continue be leading cause morbidity mortality worldwide (Benjamin et al, 2017). failure, characterized inability ventricle sufficiently pump blood, end stage various forms cardiovascular diseases, (MI), valvular disease, (Harjola 2016). Progressive loss key pathogenic factor development failure. Based morphological manifestations, three distinct types cell death, namely necrosis, possibly autophagy, occur during MI failure (Lee Gustafsson, 2009; Whelan 2010). While signal transduction pathways involved have been widely investigated, how initiate different stresses, ischemia toxic chemicals, remains unclear. Apoptosis programmed plays important progression 2009). Two classic pathways—the extrinsic intrinsic pathways—mediate apoptotic mammalian (Moe Marin-Garcia, The triggered ligands, Fas tumor necrosis factor-α. binding ligands their individual receptors leads caspase-8-mediated cascade. pathway, also called mitochondrial initiated stress, oxidative DNA damage, ultimately results membrane permeabilization, cytochrome C release, subsequent caspase-9-mediated Caspase-12 resident caspase recently identified mediate high calcium concentration low oxygen (Nakagawa Yuan, 2000; Nakagawa 2000). Ischemia can increase (Lam 2013; Xu 2015; Fu are observed tissues from patients MI, dilated cardiomyopathy, (Narula 1996; Olivetti Saraste 1997), well animal models injuries (Fliss Gattinger, Qin 2005). Pharmacological genetic diminishes infarct size improves function after (Whelan Therefore, targeting promising preventive strategy (Yang 2013a). Prostaglandin bioactive metabolite arachidonic acid produced sequential reaction cyclooxygenases (COXs) PGD2 synthases. exerts functions D-prostanoid receptor 1 (DP1) (CRTH2, named DP2). It implicated pathophysiological events, especially inflammation (Santus Radovanovic, DP1 abundantly brain tissues, mast cells, macrophages, immune cell-enriched organs (Sawyer 2002; Santus Since detectable (Katsumata 2014), PGD2/DP1 axis glucocorticoid-induced (Tokudome 2009), probably M2 macrophage-mediated timely resolution injured hearts (Kong 2016, unknown. In present study, upregulated treatment, increased Unexpectedly, deletion attenuated DOX-induced conferred mice. deficiency suppressed ER-specific Ca2+-dependent cysteine protease activity cardiomyocytes, promoted anoxia-induced activating homolog mouse caspase-12. Thus, our Results To determine whether PGs isolated primary neonatal examined alterations PG production expression PGD2, PGE2, PGF2α, TxB2 were significantly elevated (Fig 1A Appendix Fig S1A). CRTH2, IP, FP, EP1, EP4 whereas barely detected. Anoxia boosted (2.7-fold), while repressed FP EP1 1B S1B). investigate changes contribute directly caspase-3, effector caspase, these agonist-treated agonist DK-PGD2 induced caspase-3 but other agonists (Lat-FA, agonist; ONO-DI-004, misoprostol, agonist) had no overt effects 1C). Moreover, gradually time-dependent manner 1D). murine model permanent border zone day post-MI peaked 3, compared remote region 1E). These indicate anoxic stress. Figure 1. challenged h. Data represent mean ± SEM.**P < 0.0001 vs. control (Mann–Whitney U-test); n = 6. Relative mRNA levels exposed SEM. **P Western blot analysis treated condition. Lat-FA, DK-PGD2, agonist. manner. 10 min, *P 0.029, 0 min (one-way ANOVA); 30 0.000284 60 0.00016, regions post-MI. 1, 0.000765, area (unpaired two-tailed t-test); 0.0003, 7, 0.00542, data available online figure. [emmm201708237-sup-0003-SDataFig1.pdf] Download figure PowerPoint protects ischemia-induced resulted ~18.2% cultured within h; both TUNEL staining EV1A B) flow cytometric EV1C D) decreased CRTH2−/− EV1E). agreement vitro observations, reduced TUNEL+ zones 2A 2C) without affecting autophagy (Appendix S2A B), therefore improving 14 2D–F). dissection showed significant reduction WT (28.5 2.2% 19.7 1.4%, P 0.01; 2G H). lower ratio mass body weight (HW/BW, 2I), higher survival rate (87.5% versus 68.4% WT; 2J), less collagen deposition than 2K L). Similarly, blockade selective antagonist CAY10595 protected evidenced increasing EV2A–C) sizes EV2D–E). immunostaining confirmed CAY10595-treated EV2F–G). differences found capillary density (CD31+) S3A pro-angiogenic growth factors, VEGF, FGF, HGF, PDGF S3C), risk between Click expand EV1. attenuates Representative TUNEL-stained images Green, TUNEL-positive nuclei; blue, DAPI-stained red, labeled antibody α-actinin; scale bar, 50 μm. Quantification (A). 0.00202, Flow cytometry annexin V propidium iodide (PI) following treatment. V+ (C). 0.0002, 2. (MI) A. peri-infarct DAPI; B. 0.0147, WT, 6; CRTH2−/−, 8. C. D–F. M-mode echocardiographic EF, ejection fraction (D); FS, fractional shortening (E); LVPWs, left ventricular posterior wall thickness end-systole. (F). 0.0015, 0.00805, 10; 12. G. Evans blue TTC-stained Scale 500 H. infarcted MI. 0.00354, 10. I. weight-to-body subjected 0.0119, (Sham), 8; J. Kaplan–Meier curves 0.0356, (log-rank test).WT, 29; 26. K. Masson's trichrome hearts. 20 L. content (K). 0.0196, 7; 9. [emmm201708237-sup-0004-SDataFig2.pdf] EV2. A–C. Echocardiographic (5 mg/kg/day) (A); (B); systolic end-systole 0.0346, Vehicle; 0.0138, 0.0129, Vehicle Vehicle, CAY10595, D. TTC E. (D). 0.0172, vehicle F. 0.0163, Functional reprogrammed fibroblasts transcriptional factors—Gata4, Mef2c, Tbx5 (GMT; Ieda 2010; 2015). We further investigated reprogramming into using GMT system. Immunostaining troponin (cTnT)-positive (~15%) GMT, difference efficiency detected EV3A B); similar beating rates cTnT-positive transdifferentiated time points tested EV3C). Consistently, cardiomyocyte-specific genes EV3D). suggested fibroblast. EV3. Effect immunofluorescence (cTnT) (CFs) co-transfection transcription (GMT). Blue, cells; total cTnT+ SEM; (Vector), Percentage among cells. 5. Cardiac-specific gene GMT-infected CFs weeks postinfection. 0.05, vector CRTH2+/+ bone marrow (BM) reconstitution does influence repair An appropriate inflammatory required (Dutta Nahrendorf, Both macrophages (CD68+) neutrophils (Ly6G+) recruited one week test. recruitment S4A–D) related cytokines S4E) Th2 migration (Satoh 2006). retarded T-cell infiltration (CD4+) S5A (IL-4, IL-5, IL-13) inflamed S5C). explore diminished mice, reconstituted BM EV4A). Decreased restored transplantation (WT→KO), received (KO→WT) EV4B C). did failed prevent EV4D E). EV4F G). may mainly ascribed BM-derived Th2. EV4. Genotyping respective transplanted CD4+ undergoing 7 CD4-positive cell; (B). 0.0125, KO→WT WT→WT; 0.0148, KO→KO WT→KO 7. pe