Large Stress Proteins in Biology and Medicine

描述（由申请方提供）：通过热休克在体外与临床相关肿瘤蛋白抗原复合的大应激/热休克蛋白（LSP）已在动物模型中显示出高效的抗肿瘤疗效。然而，LSP的免疫调节功能的分子基础仍然在很大程度上未确定。我们最初的工作导致了一个意想不到的发现，即通过外源性递送LSP增强的黑素瘤蛋白抗原（Ag）的交叉呈递涉及内质网相关降解（ERAD）机制，一种细胞内蛋白质质量控制系统。此外，我们首次发现LSPs与病原体衍生分子或PAMPs积极相互作用，导致细胞内NOD样受体（NLR）/炎性小体的协同激活。这些意见，这涉及的分子伴侣的基本免疫活动的LSP，使我们假设，LSP能够从事和桥接先天和适应性区室通过其高效的伴侣功能。本申请的总体目标是研究LSP与自身肿瘤蛋白Ag和PAMP分子相互作用的高度新颖的方面，以及不同的免疫后果。此外，将评估在生理学相关的发热样热应激条件下LSP引发的细胞应答。作为这些研究的结果，将评估通过整合Ag靶分子和PAMP分子来进一步增强伴侣疫苗方案的治疗效力的新方法。为实现本课题的研究目标，本研究将从以下几个方面进行研究：1）确定LSP增强肿瘤蛋白Ag交叉呈递的蛋白质质量控制机制。2)确定LSP与PAMP相互作用并改变所得免疫结果的能力。3)确定结合PAMP衍生的“危险”信号的改良分子伴侣疫苗方案的抗肿瘤功效。鉴于不同类别的热休克蛋白所共有的共同伴侣特性，这些研究的成功完成将为这些古老分子的独特生物活性提供重要的见解。拟议的研究还将揭示轻度热应激/高温在宿主反应中的作用，这可以在治疗上用于免疫调节。对LSP在宿主防御机制中的作用的详细了解应该为靶向癌症和其他具有临床重要性的疾病提供更有效和安全的策略。

DESCRIPTION (provided by applicant): Large stress/heat shock proteins (LSPs) that are complexed in vitro with clinically relevant tumor protein antigens by heat shock have demonstrated highly potent antitumor efficacy in animal models. However, the molecular basis of immunoregulatory features of LSPs remains largely undetermined. Our initial work led to an unexpected finding that enhanced cross-presentation of melanoma protein antigen (Ag) by exogenously delivered LSP involved the endoplasmic reticulum-associated degradation (ERAD) machinery, an intracellular protein quality control system. In addition, for the first time we have discovered that LSPs actively interact with pathogen-derived molecules or PAMPs, resulting in synergistic activation of intracellular NOD-like receptor (NLR)/inflammasome. These observations, which implicate the molecular chaperoning underlying the essential immunologic activities of LSPs, lead us to hypothesize that LSPs are capable of engaging and bridging both innate and adaptive compartments through their highly efficient chaperoning functions. The overall objective of this application is to investigate the highly novel aspects of LSPs in interacting with a self tumor protein Ag and PAMP molecules, as well as distinct immune consequences. Furthermore, LSP-primed cellular responses under physiologically relevant fever-like thermal stress conditions will be assessed. As a consequence of these studies, a novel approach to further enhance the therapeutic potency of the chaperone vaccine regimen by integrating both Ag target and PAMP molecules will be evaluated. The following specific aims will be pursued to achieve the project goal: 1) Determine the protein quality control mechanisms underlying LSP-enhanced cross-presentation of tumor protein Ag. 2) Determine the capability of LSP to interact with PAMPs and modify the resultant immune outcomes. 3) Determine the antitumor efficacy of a modified chaperone vaccine regimen incorporating a PAMP-derived 'danger' signal. Given the common chaperoning property shared by different classes of heat shock proteins, successful completion of these studies will provide important insights into the unique biological activities of these ancient molecules. The proposed studies will also shed new light on the role of mild thermal stress/hyperthermia in the host response, which could be exploited therapeutically for immune modulation. A detailed understanding of the LSP action in host defense mechanisms should provide more effective and safe strategies for targeting cancer and other diseases of clinical importance.