光敏剂Verteporfin诱导YAP1 SUMO化对YAP1功能的调控在子宫内膜癌靶向治疗中的机理研究。

The incidence of Endometrial cancer (EC) is increasing and the age of population of EC is getting younger. To explore the new targeting drugs is the focus of current oncological studies. The clinic data show more than 90% EC has the activation of PI3K/Akt/mTOR pathway and YAP1 that is the major downstream effector of Hippo pathway could up-regulate GAB2 to active the PI3K/Akt/mTOR pathway and is closed to the malignancy and survival of EC. Therefore, YAP1 could be able to be a promising target for EC target therapy. Verteporfin was revealed as an inhibitor of YAP and we found it has an inhibited effect on proliferation and invasion of EC cells by experiments at the cellular level and animal model. in additional, the effects of VP was relieved by the decrease of YAP1. How does Verteporfin inhibit the function of YAP1, which is unknown in EC. Our previous work preliminarily found the SUMOylation of YAP1 induced by Verteporfin. We hypothesize that Verteporfin could inhibit EC by inducing the SUMOylation of YAP1, which can inhibit the function of YAP1. Based on our previous work, this study will validate our hypothesis deeply via in vivo and in vitro and explore the mechanism of SUMOylation of YAP1 by Verteporfin as well as the effects of SUMOylation of YAP1 on EC cells. It will provide the evidences that support the therapy targeting YAP1 for EC and the possibility that Verteporfin could be able to be a new promising drug for the targeting therapy of EC in the future.

子宫内膜癌（EC）发病率逐年升高且呈年轻化趋势，其靶向药物是近年来的研究焦点。临床资料显示90%以上的EC有PI3K/Akt/mTOR通路的激活，前期工作证实Hippo通路效应分子YAP1可以上调GAB2激活PI3K/Akt/mTOR通路，与EC的恶性程度及生存时间密切相关。 因此，YAP1可成为EC治疗的新靶点。Verteporfin（VP）是YAP的抑制剂，前期通过细胞实验和动物模型均证实VP具有明显抑制内膜癌细胞增殖和侵袭的作用，降调YAP1导致VP作用减弱。VP如何调节YAP1？目前尚不明了。前期发现VP诱导YAP1的SUMO化，我们推测VP诱导YAP1的SUMO化抑制YAP1的功能，从而发挥抑制EC的作用。本研究将在前期工作基础上，从体内外、多环节验证假设，揭示VP对YAP1的作用机制及其对YAP1功能的影响，为今后针对YAP1为靶点，将VP用于EC靶向治疗提供前期基础。

子宫内膜癌是女性生殖系统常见的三大恶性肿瘤之一，其发病率逐年升高。近年来研究发现子宫内膜癌主要由PI3K/Akt/mTOR通路驱动，与HIPPO/YAP通路存在密切联系。而子宫内膜癌中普遍存在YAP1激活，YAP1激活水平与子宫内膜癌的恶性程度和预后密切相关。Verteporfin 是 YAP 的特异抑制剂。我们前期工作首次报道Verteporfin 明显抑制子宫内膜癌细胞的增殖，侵袭能力，而且动物模型证实 Verteporfin 明显抑制移植瘤生长速度，降调 YAP1 水平减弱 Verteporfin 的作用，提示Verteporfin可能通过 YAP1 起到抑制子宫内膜癌的作用，但其具体调节机制尚无探究。.本项目借助蛋白免疫共沉淀、RNA测序、TCGA数据库分析等技术，发现Verteporfin对子宫内膜癌细胞具有抑制增殖和促进凋亡作用，同时使子宫内膜癌细胞的贴壁减少，迁移能力减弱，同时可以抑制子宫内膜不典型和子宫内膜癌的类器官。Verteporfin促进子宫内膜癌细胞中YAP1蛋白与SUMO1蛋白发生结合，YAP1蛋白第280位赖氨酸位点突变后，YAP1与SUMO蛋白结合减少，而YAP1蛋白第127位丝氨酸位点突变后，会抑制YAP蛋白和SUMO蛋白的结合。除了SUMO化修饰外，Verteporfin还诱导子宫内膜癌细胞发生广泛的NEDD8蛋白修饰。此外，Verteporfin抑制子宫内膜癌细胞线粒体功能，同时抑制线粒体的分布和形态，降低线粒体中酮戊二酸脱氢酶的含量。.本项目对YAP1 SUMO化的修饰机制的研究，进一步明确Verteporfin如何调节YAP1的功能，同时并靶向YAP1 SUMO化修饰的具体位点，为今后 Verteporfin 可能用于子宫内膜癌的靶向治疗提供重要的前期基础。此外，本项目首次发现Verteporfin在子宫内膜癌细胞线粒体呼吸功能中的作用进行研究，有助于理解Verteporfin在子宫内膜癌发生发展中的作用。.项目执行期间，发表高水平学术论文6篇，其中SCI收录4篇，中文核心期刊2篇。培养硕士研究生2名。