Division of Hematology and Hemostaseology, Department of Internal Medicine I, Medical University of Vienna, Vienna, Austria.
PLoS One. 2012;7(1):e29925. doi: 10.1371/journal.pone.0029925. Epub 2012 Jan 27.
The phosphoinositide 3-kinase (PI3-kinase) and the mammalian target of rapamycin (mTOR) are two major signaling molecules involved in growth and activation of mast cells (MC) and basophils (BA). We examined the effects of the dual PI3-kinase/mTOR blocker NVP-BEZ235 on growth of normal and neoplastic BA and MC as well as immunoglobulin E (IgE)-dependent cell activation. Growth of MC and BA were determined by measuring (3)H-thymidine uptake and apoptosis. Cell activation was determined in histamine release experiments and by measuring upregulation of CD63 and CD203c after challenging with IgE plus anti-IgE or allergen. We found that NVP-BEZ235 exerts profound inhibitory effects on growth of primary and cloned neoplastic MC. In the MC leukemia cell line HMC-1, NVP-BEZ235 showed similar IC(50) values in the HMC-1.1 subclone lacking KIT D816V (0.025 µM) and the HMC-1.2 subclone expressing KIT D816V (0.005 µM). Moreover, NVP-BEZ235 was found to exert strong growth-inhibitory effects on neoplastic MC in a xenotransplant-mouse model employing NMR1-Foxn1(nu) mice. NVP-BEZ235 also exerted inhibitory effects on cytokine-dependent differentiation of normal BA and MC, but did not induce growth inhibition or apoptosis in mature MC or normal bone marrow cells. Finally, NVP-BEZ235 was found to inhibit IgE-dependent histamine release in BA and MC (IC(50) 0.5-1 µM) as well as anti-IgE-induced upregulation of CD203c in BA and IgE-dependent upregulation of CD63 in MC. In summary, NVP-BEZ235 produces growth-inhibitory effects in immature neoplastic MC and inhibits IgE-dependent activation of mature BA and MC. Whether these potentially beneficial drug effects have clinical implications is currently under investigation.
磷酸肌醇 3-激酶(PI3-kinase)和哺乳动物雷帕霉素靶蛋白(mTOR)是参与肥大细胞(MC)和嗜碱性粒细胞(BA)生长和激活的两个主要信号分子。我们研究了双重 PI3-激酶/mTOR 抑制剂 NVP-BEZ235 对正常和肿瘤性 BA 和 MC 的生长以及免疫球蛋白 E(IgE)依赖性细胞激活的影响。通过测量[3H]胸腺嘧啶摄取和细胞凋亡来确定 MC 和 BA 的生长。通过组胺释放实验和在 IgE 加抗 IgE 或变应原刺激后测量 CD63 和 CD203c 的上调来确定细胞激活。我们发现 NVP-BEZ235 对原代和克隆肿瘤性 MC 的生长具有深远的抑制作用。在 MC 白血病细胞系 HMC-1 中,NVP-BEZ235 在缺乏 KIT D816V 的 HMC-1.1 亚克隆(0.025 µM)和表达 KIT D816V 的 HMC-1.2 亚克隆(0.005 µM)中显示出相似的 IC50 值。此外,NVP-BEZ235 在使用 NMR1-Foxn1(nu) 小鼠的异种移植小鼠模型中对肿瘤性 MC 也具有强烈的生长抑制作用。NVP-BEZ235 还对正常 BA 和 MC 的细胞因子依赖性分化产生抑制作用,但在成熟 MC 或正常骨髓细胞中不诱导生长抑制或细胞凋亡。最后,发现 NVP-BEZ235 抑制 IgE 依赖性 BA 和 MC 中的组胺释放(IC50 0.5-1 µM)以及抗 IgE 诱导的 BA 中的 CD203c 上调和 IgE 依赖性 MC 中的 CD63 上调。总之,NVP-BEZ235 对不成熟的肿瘤性 MC 产生生长抑制作用,并抑制 IgE 依赖性成熟 BA 和 MC 的激活。这些潜在的有益药物作用是否具有临床意义目前正在研究中。