Tumor-Targeted Synergistic Blockade of MAPK and PI3K from a Layer-by-Layer Nanoparticle.
Tumor-Targeted Synergistic Blockade of MAPK and PI3K from a Layer-by-Layer Nanoparticle.
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DOI：
10.1158/1078-0432.ccr-15-0013
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发表时间：
2015-10-01
期刊：
影响因子：
--
通讯作者：
Hammond PT
中科院分区：
文献类型：
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作者：
Dreaden EC;Kong YW;Morton SW;Correa S;Choi KY;Shopsowitz KE;Renggli K;Drapkin R;Yaffe MB;Hammond PT
RAS/RAF/MEK/ERK和PI 3 K/AKT/mTOR细胞信号传导通路之间的串扰和反馈对于各种实体瘤中的肿瘤起始、维持和对靶向治疗的适应性抗性至关重要。这些通路的联合阻断-水平阻断-是一种有前景的治疗策略;然而，游离小分子抑制剂组合的复合剂量限制性毒性是其临床应用的显著障碍。AZD 6244（司美替尼），一种Mek 1/2的变构抑制剂，和PX-866，一种PI 3 K的共价抑制剂，被共封装在肿瘤靶向纳米级药物制剂-逐层（LbL）纳米颗粒中。除了体外细胞进入、协同细胞杀伤和组合信号阻断之外，还表征了纳米级制剂的结构、尺寸和表面电荷。通过活体动物荧光/生物发光成像、蛋白质印迹、血清细胞因子分析和免疫组织化学，在携带乳腺肿瘤异种移植物的NCR裸鼠中研究体内肿瘤靶向和治疗。来自纳米级制剂（160 ± 20 nm，−40 ± 1 mV）的MAPK和PI 3 K轴联合阻断在体外对三阴性乳腺癌（MDA-MB-231）和RAS突变型肺肿瘤细胞（KP 7 B）具有协同毒性，这种作用在包封后进一步增强。在体内，全身施用的LbL纳米颗粒优先靶向皮下MDA-MB-231肿瘤异种移植物，同时阻断末端激酶Erk和Akt的肿瘤特异性磷酸化，并且在不存在从游离药物组合观察到的剂量限制性肝毒性作用的情况下引起显著的疾病稳定。接受非靶向但双载药纳米颗粒的小鼠表现出进行性疾病。肿瘤靶向纳米级药物制剂可以为临床上协同阻断MAPK和PI 3 K提供更安全有效的手段。
Cross-talk and feedback between the RAS/RAF/MEK/ERK and PI3K/AKT/mTOR cell signaling pathways is critical for tumor initiation, maintenance, and adaptive resistance to targeted therapy in a variety of solid tumors. Combined blockade of these pathways—horizontal blockade—is a promising therapeutic strategy; however, compounded dose-limiting toxicity of free small molecule inhibitor combinations is a significant barrier to its clinical application. AZD6244 (selumetinib), an allosteric inhibitor of Mek1/2, and PX-866, a covalent inhibitor of PI3K, were co-encapsulated in a tumor-targeting nanoscale drug formulation—layer-by-layer (LbL) nanoparticles. Structure, size, and surface charge of the nanoscale formulations were characterized, in addition to in vitro cell entry, synergistic cell killing, and combined signal blockade. In vivo tumor targeting and therapy was investigated in breast tumor xenograft-bearing NCR nude mice by live animal fluorescence/bioluminescence imaging, Western blotting, serum cytokine analysis, and immunohistochemistry. Combined MAPK and PI3K axis blockade from the nanoscale formulations (160 ± 20 nm, −40 ± 1 mV) was synergistically toxic toward triple-negative breast (MDA-MB-231) and RAS-mutant lung tumor cells (KP7B) in vitro, effects that were further enhanced upon encapsulation. In vivo, systemically administered LbL nanoparticles preferentially targeted subcutaneous MDA-MB-231 tumor xenografts, simultaneously blocked tumor-specific phosphorylation of the terminal kinases Erk and Akt, and elicited significant disease stabilization in the absence of dose-limiting hepatotoxic effects observed from the free drug combination. Mice receiving untargeted, but dual drug-loaded nanoparticles exhibited progressive disease. Tumor-targeting nanoscale drug formulations could provide a more safe and effective means to synergistically block MAPK and PI3K in the clinic.