HSP60, Inflammation and Cardiovascular Disease

描述（由申请人提供）：以前我们已经表明，细胞外（ex）热休克蛋白（HSP）60导致心肌细胞凋亡和TNF和IL-1的合成。阻断Toll样受体（TLR）4减少，但不能消除凋亡，表明HSP 60与TLR 4的相互作用是复杂的，涉及其他蛋白质。我们的过度假设是，HSP 60可以引起细胞凋亡和炎症，这与HSP 60的异常表达、分布或翻译后修饰有关。我们将在这一重要领域扩大我们的研究，有3个具体的目标，旨在进一步确定HSP 60在心力衰竭和其他心血管疾病中的作用。SA 1-研究exHSP 60介导的细胞凋亡和炎症的机制。我们将扩大我们的调查TLR 4 exHSP 60信号，确定是否，TLR 4抑制剂，在体外和体内，可以减少细胞凋亡，并进一步表征质膜相关HSP 60。SA 2-研究细胞外HSP 60运输和翻译后修饰在其毒性中的作用。我们推测HSP 60的翻译后修饰影响其定位和细胞外毒性。研究发现，糖尿病和其他疾病患者的血液中存在高水平的HSP 60。我们将研究翻译后修饰以及外泌体在这种HSP 60毒性中的作用。其次，我们将研究膜相关的HSP 60的来源。SA 3-研究HSP 60在重复性损伤和热休克中的作用-热休克反应是保护性的，然而，当热休克反应在炎症应激后诱导时，而不是保护，因此发生损伤增加。我们发现，热休克后，炎症热休克蛋白60增加，但不是当这些刺激的顺序是颠倒的。HSP 60的增加与细胞凋亡的增加有关。此外，阻断活化的热休克因子（HSF）可减少细胞凋亡。我们将研究HSF 1和2在选择性敲除模型中的热休克悖论中的作用。HSP 60在热休克悖论中的作用将通过检查其细胞定位和操纵其表达来研究。计划中的工作将扩大PI对HSP 60及其在心血管炎症和细胞凋亡中的作用的研究，并有可能导致心力衰竭的新治疗方法。

DESCRIPTION (provided by applicant): Previously we have shown that extracellular (ex) heat shock protein (HSP)60 causes cardiac myocyte apoptosis and the synthesis of TNF and IL-1$. Blocking toll-like receptor (TLR)4 decreased, but did not abolish apoptosis, suggesting that the interaction of HSP60 with TLR4 is complex and involves other proteins. Our over-arching hypothesis is that HSP60 can cause apoptosis and inflammation, and this is associated with abnormal expression, distribution or post-translational modification of HSP60. We will expand our investigation in this important area with 3 specific aims designed to further define the role of HSP60 in heart failure and other cardiovascular disease. SA1 - Investigate the mechanism(s) of exHSP60 mediated apoptosis and inflammation. We will extend our investigation of TLR4 in exHSP60 signaling, determine whether, Eritoran, an inhibitor of TLR4, can reduce apoptosis in vitro and in vivo, and further characterize plasma membrane associated HSP60. SA2 - Investigate extracellular HSP60 trafficking and the role of post-translational modification in its toxicity. We hypothesize that post-translational modifications of HSP60 influence its localization and its extracellular toxicity. Studies have found high levels of HSP60 in the blood of patients with diabetes and other diseases. We will investigate the role of post- translational modifications as well as exosomes in the toxicity of this HSP60. Second we will examine the source of membrane associated HSP60. SA3 - Investigate the role of HSP60 in Repetitive Injury and the Heat Shock Paradox - The heat shock response is protective, and yet, when the heat shock response is induced after an inflammatory stress, rather than protection, increased injury occurs, hence. We found that HSP60 is increased after inflammation followed by heat shock, but not when the order of these stimuli is reversed. This increase in HSP60 is associated with increased apoptosis. In addition, blocking activated heat shock factor (HSF) decreased apoptosis. We will investigate the role of HSF1 and 2 in the heat shock paradox with selective knockout models. HSP60's role in the heat shock paradox will be studied by examining its cellular localization and manipulating its expression. The planned work will expand the PI's investigation of HSP60 and its role in cardiovascular inflammation and apoptosis and has the potential to lead to new therapeutic approaches to heart failure.