子宫癌肉瘤（UCS）是罕见的、具有生物学侵袭性的肿瘤。由于 UCS 可能存在 RAS/MAPK 通路基因突变，我们评估了 RAF/MEK 钳 avutometinib 联合粘着斑激酶 (FAK) 抑制剂 defactinib 或 VS-4718 对多种原代 UCS 细胞系的临床前体外和体内疗效全外显子组测序 (WES) 用于评估 5 个原代 UCS 细胞系的遗传景观。使用针对原代 UCS 细胞系的细胞活力和细胞周期测定来评估 avutometinib ± FAK 抑制剂的体外活性。使用蛋白质印迹分析进行机制研究，同时在通过口服强饲法接受 avutometinib ± FAK 抑制剂治疗的 UCS 荷瘤小鼠中完成体内实验。WES 结果表明多个 UCS 细胞系存在遗传改变，包括 KRAS、PTK2、BRAF、MAP2K 和 MAP2K1 ，可能对 FAK 和 RAF/MEK 抑制敏感。当单独或组合使用时，五分之四的 UCS 细胞系在体外表现出对 FAK 和/或 RAF/MEK 抑制的敏感性。通过蛋白质印迹测定，将 UCS 细胞系暴露于 defactinib/avutometinib 组合后证明磷酸化 (p)-FAK 和 p-ERK 均减少。在体内，在 UCS 异种移植物中，与单药治疗和对照从第 10 天开始 (p < 0.002) 相比，avutometinib/VS-4718 组合表现出优异的肿瘤生长抑制作用和更长的生存期。avutometinib 和 defactinib 的组合在体外和临床试验中表现出良好的前景。针对原代 UCS 细胞系和异种移植物的体内抗肿瘤活性。版权所有 © 2024 Elsevier Inc. 保留所有权利。

Uterine carcinosarcomas (UCS) are rare, biologically aggressive tumors. Since UCS may harbor mutations in RAS/MAPK pathway genes we evaluated the preclinical in vitro and in vivo efficacy of the RAF/MEK clamp avutometinib in combination with the focal adhesion kinase (FAK) inhibitors defactinib or VS-4718 against multiple primary UCS cell lines and xenografts.Whole-exome-sequencing (WES) was used to evaluate the genetic landscape of 5 primary UCS cell lines. The in vitro activity of avutometinib ± FAK inhibitor was evaluated using cell viability and cell cycle assays against primary UCS cell lines. Mechanistic studies were performed using western blot assays while in vivo experiments were completed in UCS tumor bearing mice treated with avutometinib ± FAK inhibitor by oral gavage.WES results demonstrated multiple UCS cell lines harbor genetic alterations including KRAS, PTK2, BRAF, MAP2K, and MAP2K1, potentially sensitizing to FAK and RAF/MEK inhibition. Four out of five of the UCS cell lines demonstrated in vitro sensitivity to FAK and/or RAF/MEK inhibition when used alone or in combination. By western blot assays, exposure of UCS cell lines to the combination of defactinib/avutometinib demonstrated decreased phosphorylated (p)-FAK as well as decreased p-ERK. In vivo, the combination of avutometinib/VS-4718 demonstrated superior tumor growth inhibition and longer survival compared to single agent treatment and controls starting at day 10 (p < 0.002) in UCS xenografts.The combination of avutometinib and defactinib demonstrates promising in vitro and in vivo anti-tumor activity against primary UCS cell lines and xenografts.Copyright © 2024 Elsevier Inc. All rights reserved.