熱休克蛋白90抑制劑AUY922對於發炎反應的免疫調控

Immunomodulation of HSP90 inhibitor NVP-AUY922 on the inflammatory response

賈志元(Chih-Yuan Chia)

指導教授 ： 呂思潔

臺北醫學大學/醫學院/醫學科學研究所/碩士(2014年)

摘要

熱休克蛋白90是一種伴隨蛋白，經由與目標蛋白結合後幫助目標蛋白摺疊成正確的蛋白質結構，可以調控細胞週期、自我凋亡和訊息傳遞，這些都跟癌症有關，例如：細胞存活、增生、侵犯、轉移及血管新生，所以目前有多種Hsp90抑制物正在人體試驗，評估是否能治療癌症。本篇所研究的AUY922是一種熱休克蛋白90的抑制劑，會與熱休克蛋白90α上的ATP 結合位結合，以達到抑制的作用。已知AUY922可減低多種癌症腫瘤大小，例如：胃癌（gastric cancer）、多發性骨髓瘤（multiple myeloma）、乳癌（breast cancer）、前列腺癌（prostate cancer）及實質固態瘤（solid tumor）等，而AUY922正在人體試驗中被研究是否能治療這些癌症。
過去的文獻中指出熱休克蛋白90抑制劑會抑制發炎反應中的訊息傳遞因子，減少前發炎激素的表現量，藉此降低發炎反應。本篇利用PMA誘導THP-1細胞分化為巨噬細胞，再用LPS活化巨噬細胞，藉此觀察AUY922對巨噬細胞以及巨噬細胞活化過程的影響。我們使用沒有細胞毒性的AUY922濃度，在巨噬細胞及活化過程中發現，AUY922能有意義的減少前發炎激素IL-1β表現量，但不影響IL-8、IL-10、TNF-α的表現量，也會增加熱休克蛋白70表現量，但磷酸化的GSK-3（Ser21/9）則無明顯變化，也不會影響活性氧化物的表現量。在巨噬細胞活化的過程中，AUY922可以減少MMP-9的分泌量，但不會影響MMP-2的分泌量，也不會影響巨噬細胞的細胞表面分子。在敗血症小鼠模式中AUY922會減少IL-6、IL-10、IL-12p70、IFN-γ、MCP-1的表現量，也會減少體重下降的幅度。總之，AUY922在細胞及小鼠中都能減少發炎反應的產生。

關鍵字

熱休克蛋白90 ； AUY922 ； 發炎反應 ； 巨噬細胞

並列摘要

Heat shock protein 90(Hsp90) is a kind of chaperone, binding client protein to help it to fold the correct protein structure. Hsp90 maintains cell cycle, apotosis and signal transduction. These are all about cancer, for example: cell viability, proliferation, invasion, migration and angiogenesis. Now, many Hsp90 inhibitors try to therapy cancer in clinical trial. In the study, AUY922 is the Hsp90 inhibitor and binds ATP-binding site of Hsp90α to inhibit Hsp90. It can decrease the size of tumor in many cancers, example: gastric cancer, multiple myeloma, breast cancer, prostate cancer and solid tumor. In previous study, Hsp90 inhibitor can inhibit signal transduction factor of inflammation and reduce expression of pro-inflammatory cytokine for decreasing inflammation. This study used PMA to induce THP-1 cell differentiation to macrophage, and then use lipopolysaccharide to induce macrophage activation. We observe the infleunce of AUY922 in PMA-induced macrophage and LPS-activated macrophage. We treat cells with non-cytotoxic concentration of AUY922 in this experiment. We found that AUY922 decreased expression of IL-1β, but no influence in expression of IL-8, IL-10, TNF-α in PMA-induced macrophage and LPS-activated macrophage. AUY922 increased expression of Hsp70, but no influence in expression of p-GSK-3(Ser21/9) and reactive oxygen species. In LPS- activated macrophage, AUY922 decreased expression of extracellular MMP-9 but not MMP-2. In sepsis mice model, AUY922 decrease expression of IL-6, IL-10, IL-12p70, IFN-γ, MCP-1 and range of mice weight decline. In conclusion, AUY922 may decrease inflammation in vitro and in vivo.

並列關鍵字

hsp90 ； AUY922 ； inflammatory response ； macrophage

參考文獻

1. F. Ritossa. A new puffing pattern induced by temperature shock and DNP in drosophila. Experientia. 18:571-573 (1962).

2. M.P. Goetz, D.O. Toft, M.M. Ames, and C. Erlichman. The Hsp90 chaperone complex as a novel target for cancer therapy. Ann Oncol. 14:1169-1176 (2003).

3. M. Akerfelt, R.I. Morimoto, and L. Sistonen. Heat shock factors: integrators of cell stress, development and lifespan. Nature reviews Molecular cell biology. 11:545-555 (2010).

4. K. Bedardand K.H. Krause. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiol Rev. 87:245-313 (2007).

5. M. Taipale, D.F. Jarosz, and S. Lindquist. HSP90 at the hub of protein homeostasis: emerging mechanistic insights. Nature reviews Molecular cell biology. 11:515-528 (2010).