背景:高级别子宫内膜癌(EAC)是侵袭性肿瘤,治疗后进展风险高。由于EAC可能在RAS/MAPK通路中存在突变,我们评估了阿伏美替尼的临床前体外和体内疗效,RAF/MEK卡箍,与粘着斑激酶(FAK)抑制剂defactinib或VS-4718组合,针对多种原代EAC细胞系和异种移植物。
方法:使用全外显子组测序(WES)评估五个原代EAC细胞系的遗传格局。使用细胞活力评估了阿伏美替尼和defactinib作为单一药物和组合的体外活性,细胞周期,和细胞毒性测定。使用Western印迹测定法进行机理研究,同时在用任一载体处理的UTE10移植小鼠中完成体内实验。阿伏美替尼,VS-4718或其组合通过口服管饲法。
结果:WES结果显示多个EAC细胞系在RAS/MAPK通路中包含遗传紊乱,包括KRAS/PTEN/PIK3CA/BRAF/ARID1A,可能对FAK和RAF/MEK抑制敏感。五个EAC细胞系中的五个证明了对FAK和/或RAF/MEK抑制的体外敏感性。通过蛋白质印迹分析,EAC细胞系暴露于defactinib,阿伏美替尼,和它们的组合显示减少的磷酸化FAK(p-FAK)以及减少的p-MEK和p-ERK。在UTE10异种移植物中,在第9天开始,与单一药剂治疗和对照相比,阿伏美替尼/VS-4718的组合在体内表现出优异的肿瘤生长抑制(p<0.02和p<0.04)。
结论:阿伏美替尼,defactinib,在更大程度上它们的组合,证明了对EAC细胞系和异种移植物的体外和体内活性。这些临床前数据支持这种组合在高级别EAC患者中的潜在临床评估。
BACKGROUND: High-grade endometrial cancers (EAC) are aggressive tumors with a high risk of progression after treatment. As EAC may harbor mutations in the RAS/MAPK pathways, we evaluated the preclinical in vitro and in vivo efficacy of avutometinib, a RAF/MEK clamp, in combination with the focal adhesion kinase (FAK) inhibitors defactinib or VS-4718, against multiple primary EAC cell lines and xenografts.
METHODS: Whole-exome sequencing (WES) was used to evaluate the genetic landscape of five primary EAC cell lines. The in vitro activity of avutometinib and defactinib as single agents and in combination was evaluated using cell viability, cell cycle, and cytotoxicity assays. Mechanistic studies were performed using Western blot assays while in vivo experiments were completed in UTE10 engrafted mice treated with either vehicle, avutometinib, VS-4718, or their combination through oral gavage.
RESULTS: WES results demonstrated multiple EAC cell lines to harbor genetic derangements in the RAS/MAPK pathway including KRAS/PTEN/PIK3CA/BRAF/ARID1A, potentially sensitizing to FAK and RAF/MEK inhibition. Five out of five of the EAC cell lines demonstrated in vitro sensitivity to FAK and/or RAF/MEK inhibition. By Western blot assays, exposure of EAC cell lines to defactinib, avutometinib, and their combination demonstrated decreased phosphorylated FAK (p-FAK) as well as decreased p-MEK and p-ERK. In vivo the combination of avutometinib/VS-4718 demonstrated superior tumor growth inhibition compared to single-agent treatment and controls starting at Day 9 (p < 0.02 and p < 0.04) in UTE10 xenografts.
CONCLUSIONS: Avutometinib, defactinib, and to a larger extent their combinations, demonstrated promising in vitro and in vivo activity against EAC cell lines and xenografts. These preclinical data support the potential clinical evaluation of this combination in high-grade EAC patients.