François Rony A, Maeng Kyungah, Nawab Akbar, Kaye Frederic J, Hochwald Steven N, Zajac-Kaye Maria
Department of Anatomy and Cell Biology, University of Florida College of Medicine, Gainesville, FL (RAF, KM, AN, MZK); Department of Medicine, University of Florida College of Medicine, Gainesville, FL (FJK); Department of Surgical Oncology, Roswell Park Cancer Institute, Buffalo, NY (SNH).
J Natl Cancer Inst. 2015 May 12;107(8). doi: 10.1093/jnci/djv123. Print 2015 Aug.
Focal adhesion kinase (FAK) mediates survival of normal pancreatic islets through activation of AKT. Upon malignant transformation of islet cells into pancreatic neuroendocrine tumors (PanNETs), AKT is frequently overexpressed and mutations in the AKT/mTOR pathway are detected. Because mTOR inhibitors rarely induce PanNET tumor regression, partly because of feedback activation of AKT, novel combination strategies are needed to target FAK/AKT/mTOR signaling.
We characterized the activation of FAK in PanNETs using immunohistochemistry and Western blot analysis and tested the FAK inhibitor PF-04554878 in human PanNET cells in vitro and in vivo (at least three mice per group). In addition, we evaluated the effect of combined FAK and mTOR inhibition on PanNET viability and apoptosis. All statistical tests were two-sided.
We found that FAK is overexpressed and hyperphosphorylated in human PanNETs and that PF-04554878 strongly inhibited FAK (Tyr397) autophosphorylation in a dose-dependent manner. We found that PF-04554878 inhibited cell proliferation and clonogenicity and induced apoptosis in PanNET cells. Moreover, oral administration of PF-04554878 statistically significantly reduced tumor growth in a patient-derived xenograft model of PanNET (P = .02) and in a human PanNET xenograft model of peritoneal carcinomatosis (P = .03). Importantly, PF-04554878 synergized with the mTOR inhibitor everolimus by preventing feedback AKT activation.
We demonstrate for the first time that FAK is overexpressed in PanNETs and that inhibition of FAK activity induces apoptosis and inhibits PanNET proliferation. We found that the novel FAK inhibitor PF-04554878 synergizes with everolimus, a US Food and Drug Administration-approved agent for PanNETs. Our findings warrant the clinical investigation of combined FAK and mTOR inhibition in PanNETs.
粘着斑激酶(FAK)通过激活AKT介导正常胰岛的存活。当胰岛细胞恶性转化为胰腺神经内分泌肿瘤(PanNETs)时,AKT经常过度表达,并且在AKT/mTOR通路中检测到突变。由于mTOR抑制剂很少能诱导PanNET肿瘤消退,部分原因是AKT的反馈激活,因此需要新的联合策略来靶向FAK/AKT/mTOR信号传导。
我们使用免疫组织化学和蛋白质印迹分析来表征PanNETs中FAK的激活情况,并在体外和体内(每组至少三只小鼠)的人PanNET细胞中测试FAK抑制剂PF-04554878。此外,我们评估了联合抑制FAK和mTOR对PanNET活力和凋亡的影响。所有统计检验均为双侧检验。
我们发现FAK在人PanNETs中过度表达且高度磷酸化,并且PF-04554878以剂量依赖性方式强烈抑制FAK(Tyr397)的自磷酸化。我们发现PF-04554878抑制PanNET细胞的增殖和克隆形成能力并诱导其凋亡。此外,口服PF-04554878在PanNET的患者来源异种移植模型(P = 0.02)和腹膜癌的人PanNET异种移植模型(P = 0.03)中均使肿瘤生长在统计学上显著降低。重要的是,PF-04554878通过防止AKT的反馈激活与mTOR抑制剂依维莫司协同作用。
我们首次证明FAK在PanNETs中过度表达,并且抑制FAK活性可诱导凋亡并抑制PanNET增殖。我们发现新型FAK抑制剂PF-04554878与依维莫司协同作用,依维莫司是一种美国食品药品监督管理局批准用于治疗PanNETs的药物。我们的研究结果值得对联合抑制FAK和mTOR在PanNETs中的应用进行临床研究。