转录因子HNF-1/4alpha调控的UGT1A3对肺腺癌IGF-1R靶向治疗的影响与机制研究

IGF-1R signaling pathway plays an important role in the tumorigenesis and development of lung adenocarcinoma, IGF-1R targeted therapy has significant effects on a variety of tumors but poor curative effects in lung cancer. In the exploration of IGF-1R target therapy, we found that intervention of downstream metabolism pathway could enhance the efficacy of IGF-1R inhibitors; in the metabolism related pathways screening, UGT1A3 was found significantly high expressed and correlated with poor prognosis in lung adenocarcinoma patients; the results of preliminary studies confirmed inhibition of UGT1A3 remarkably enhanced the antitumor effects of IGF-1R inhibitor and HNF-1/4alpha could be the potential transcriptional factor. Therefore, we speculated that the high expression of UGT1A3 regulated by HNF-1/4alpha could be an important reason for the drug resistance of IGF-1R targeted therapy, which has not been reported at home or abroad. In this study, we will use experimental techniques in vivo (tumor model in nude mice, in vivo RNA interference and imaging) and in vitro (proteomics methods, gene editing, luciferase report gene etc.) and bioinformatics analysis to clarify the effects and mechanisms of UGT1A3 on IGF-1R targeted therapy in lung adenocarcinoma from the integral, cellular and molecular levels, and reveal the transcriptional regulation mechanisms of HNF-1/4alpha on UGT1A3, hoping to provide new ideas for targeted therapy researches in lung adenocarcinoma.

IGF-1R信号通路在肿瘤发生发展中具有重要作用，IGF-1R靶向治疗对多种肿瘤疗效显著但在肺癌中疗效不佳。我们前期研究发现干预下游代谢相关通路能增强IGF-1R抑制剂疗效；对代谢通路筛选发现UGT1A3在肺腺癌中高表达且与患者不良预后显著相关；预实验证实抑制UGT1A3对IGF-1R抑制剂具有显著增效作用，HNF-1/4alpha为其转录因子。据此我们推测HNF-1/4alpha调控的UGT1A3高表达是肺腺癌IGF-1R靶向疗效不佳的重要原因，国内外对此尚无报道。本研究拟采用体内实验（裸鼠肿瘤模型，体内RNA干扰，小动物活体成像）、体外实验（蛋白质组学，基因编辑，荧光素酶报告基因等技术）和生信分析相结合的方法，从整体、细胞和分子水平阐明UGT1A3对肺腺癌IGF-1R靶向治疗的影响及机制，并深入揭示HNF-1/4alpha对UGT1A3的转录调控机制，为肺腺癌靶向治疗研究提供新思路。

肺癌是当今发病率和死亡率最高的恶性肿瘤，其分子标记和潜在治疗靶点探索一直是相关研究热点。我们前期研究发现葡萄糖醛酸转移酶1A3（UGT1A3）在肺腺癌中表达显著增高，并且其高表达与患者生存率降低显著相关。本项目中，我们对UGT1A3的转录因子进行了进一步的筛选和验证，通过研究发现，原预计调控因子HNF-1/4alpha对UGT1A3转录调控能力不强，通过生信分析及荧光素酶实验验证，我们发现转录因子上游刺激因子1（USF1）对UGT1A3的表达具有显著调控作用，目前国内外尚无相关报道。. 研究过程中，我们发现USF1、UGT1A3在肺腺癌患者广泛高表达，与患者预后显著相关；在组织水平和细胞水平验证了USF1、UGT1A3在肺腺癌中高表达，且其表达具有相关性；siRNA干扰USF1，肺腺癌细胞UGT1A3表达降低，且其增殖能力、侵袭和迁移能力受到显著抑制；体内成瘤实验证实，干扰USF1，肺腺癌细胞体内增殖受到显著抑制。我们进一步通过生信分析、表达谱芯片验证，发现neurotrophin神经营养素信号轴 (P75NTR, RIPK2, IRAK1, TRAF5 and IKKβ)在USF1对UGT1A3的调控中发挥重要作用。. 本课题研究过程按照原研究方案的不同研究层次和技术路线顺利实施，并取得了满意的研究成果。研究成果主要以SCI论文形式呈现，项目负责人董伟副教授发表相关SCI论文5篇，接收并online 1篇。为肺腺癌潜在治疗靶点USF1的研究，提供了理论和实验基础。