PI3K/AKT/mTOR inhibition in combination with doxorubicin is an effective therapy for leiomyosarcoma.
PI3K/AKT/mTOR inhibition in combination with doxorubicin is an effective therapy for leiomyosarcoma.
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DOI：
10.1186/s12967-016-0814-z
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发表时间：
2016-03-08
影响因子：
7.4
通讯作者：
Gladdy RA
中科院分区：
文献类型：
--
作者：
Babichev Y;Kabaroff L;Datti A;Uehling D;Isaac M;Al-Awar R;Prakesch M;Sun RX;Boutros PC;Venier R;Dickson BC;Gladdy RA
平滑肌肉瘤（LMS）是一种常见的软组织肉瘤，对标准化疗反应较差。因此，本研究的目的是确定新的选择性治疗，可能是有效的平滑肌肉瘤筛选细胞系的小分子库组成的480激酶抑制剂，功能上确定哪些信号通路可能是至关重要的LMS生长。用OICR激酶文库筛选LMS细胞系，并使用细胞活力测定来鉴定潜在有效的化合物。对前10%的命中进行二次验证以确定其EC 50，并进行免疫印迹以确认选择性药物作用。使用Calcusyn程序分析了用三种给药方案之一治疗后与多柔比星（Dox）的体外联合药物治疗的功效：同时治疗、用选择性化合物随后Dox的初始治疗或用Dox随后选择性化合物的初始治疗。然后使用LMS异种移植物在体内验证单一和组合药物疗法。靶向PI 3 K/AKT/mTOR途径的化合物（52%）最有效。测定EC 50以验证这些初始命中，并且在11个确认的命中中，10个靶向PI 3 K和/或mTOR途径的EC 50值<1 μM。因此，我们研究了BEZ 235和BKM 120，这两种在这些途径中的选择性化合物，是否会在体外抑制平滑肌肉瘤的生长。免疫印迹证实了这些化合物在PI 3 K和/或mTOR途径中的靶向作用。我们接下来研究了这些药物与一线化疗阿霉素（Dox）是否存在协同作用，这将允许更早地引入患者护理。BEZ 235和Dox的联合治疗在体外具有协同作用。为了在临床前模型中验证这些发现，平滑肌肉瘤异种移植物用单药和联合治疗进行治疗。BEZ 235处理的异种移植物（n = 8）显示肿瘤体积减少42%，而与单独的媒介物相比，BEZ 235与Dox组合（n = 8）使肿瘤体积减少68%。总之，本研究支持进一步研究PI 3 K和mTOR抑制剂单独使用以及与平滑肌肉瘤患者标准治疗联合使用。本文的在线版本（doi：10.1186/s12967-016-0814-z）包含补充材料，可供授权用户使用。
Leiomyosarcoma (LMS) is a common type of soft tissue sarcoma that responds poorly to standard chemotherapy. Thus the goal of this study was to identify novel selective therapies that may be effective in leiomyosarcoma by screening cell lines with a small molecule library comprised of 480 kinase inhibitors to functionally determine which signalling pathways may be critical for LMS growth. LMS cell lines were screened with the OICR kinase library and a cell viability assay was used to identify potentially effective compounds. The top 10 % of hits underwent secondary validation to determine their EC50 and immunoblots were performed to confirm selective drug action. The efficacy of combination drug therapy with doxorubicin (Dox) in vitro was analyzed using the Calcusyn program after treatment with one of three dosing schedules: concurrent treatment, initial treatment with a selective compound followed by Dox, or initial treatment with Dox followed by the selective compound. Single and combination drug therapy were then validated in vivo using LMS xenografts. Compounds that targeted PI3K/AKT/mTOR pathways (52 %) were most effective. EC50s were determined to validate these initial hits, and of the 11 confirmed hits, 10 targeted PI3K and/or mTOR pathways with EC50 values <1 μM. We therefore examined if BEZ235 and BKM120, two selective compounds in these pathways, would inhibit leiomyosarcoma growth in vitro. Immunoblots confirmed on-target effects of these compounds in the PI3K and/or mTOR pathways. We next investigated if there was synergy with these agents and first line chemotherapy doxorubicin (Dox), which would allow for earlier introduction into patient care. Only combined treatment of BEZ235 and Dox was synergistic in vitro. To validate these findings in pre-clinical models, leiomyosarcoma xenografts were treated with single agent and combination therapy. BEZ235 treated xenografts (n = 8) demonstrated a decrease in tumor volume of 42 % whereas combining BEZ235 with Dox (n = 8) decreased tumor volume 68 % compared to vehicle alone. In summary, this study supports further investigation into the use of PI3K and mTOR inhibitors alone and in combination with standard treatment in leiomyosarcoma patients. The online version of this article (doi:10.1186/s12967-016-0814-z) contains supplementary material, which is available to authorized users.