HSP60, Inflammation and Cardiovascular Disease

描述(由申请人提供)：以前我们已经证明细胞外(EX)热休克蛋白(HSP)60导致心肌细胞凋亡以及肿瘤坏死因子和白介素1的合成。阻断Toll样受体(TLR4)减少，但不能消除细胞凋亡，提示HSP60与TLR4的相互作用是复杂的，涉及其他蛋白质。我们的总体假设是，HSP60可以引起细胞凋亡和炎症，这与HSP60的异常表达、分布或翻译后修饰有关。我们将扩大我们在这一重要领域的研究，有3个具体目标，旨在进一步确定HSP60在心力衰竭和其他心血管疾病中的作用。SA1-探讨exHSP60介导细胞凋亡和炎症的机制(S)。我们将扩大我们对TLR4在exHSP60信号转导中的研究，确定TLR4的抑制剂厄立特里亚是否能在体外和体内减少细胞凋亡，并进一步鉴定质膜相关的HSP60。SA2-研究细胞外HSP60的运输和翻译后修饰在其毒性中的作用。我们假设HSP60的翻译后修饰会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HSP60水平很高。我们将研究翻译后修饰以及外切体在该HSP60毒性中的作用。其次，我们将考察膜相关HSP60的来源。SA3-研究HSP60在重复损伤中的作用和热休克悖论-热休克反应是保护性的，但是，当炎症应激后诱导热休克反应而不是保护时，损伤会增加，因此。我们发现，在炎症和热休克之后，HSP60会增加，但当这些刺激的顺序颠倒时不会。HSP60的这种增加与细胞凋亡的增加有关。此外，阻断激活的热休克因子(HSF)可减少细胞凋亡。我们将使用选择性基因敲除模型来研究HSF1和HSF2在热休克悖论中的作用。热休克蛋白60的S在热休克悖论中的作用将通过检测其细胞定位和调控其表达来研究。这项计划中的工作将扩大PI对HSP60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.