栏目寄语
胆囊癌是胆道系统最常见的恶性肿瘤，具有症状隐匿、发展迅速、早期转移、预后极差的特点，被称为新的“癌中之王”。我国是胆囊癌的高发地区之一，发病率和病死率近年来都呈持续缓慢上升趋势。胆囊慢性炎症、胆囊结石、胆囊息肉等都是胆囊癌的危险因素，但是胆囊癌目前仍缺乏特异性和敏感性都较好的早期诊断手段，临床发现的胆囊癌多为中晚期。尽管医学科技不断发展，根治性手术切除仍是当前唯一可能治愈胆囊癌的手段，行之有效的系统性治疗方法依然还在不断探索寻找中。因此，深入开展胆囊癌的临床和基础研究能够帮助我们更好地应对此类恶性肿瘤。
上海交通大学医学院附属新华医院普外科肝胆胰中心长期以来致力于胆道疾病诊治难点的攻关，形成了普外科、上海市胆道疾病研究重点实验室、上海市胆道疾病研究中心、Ⅰ期临床研究病房等多位一体的转化医学研究体系。尤其在胆囊癌的基础和临床研究方面的科研水平位于国内外领先水准，相关成果发表SCI论文50余篇，其中包括NatureGenetics、GUT、Hepatology等国际顶尖学术期刊，并获得了包括科技部新药创制项目、国家自然科学基金重点项目、上海市启明星、浦江人才、上海市优秀学科带头人、上海市卫健委新优靑等在内的多项科研及人才项目。
本期关于胆囊癌的研究主要有华西医院胆道外科李富宇教授团队发表在Molecular Cancer上关于外泌体运输的lncRNA TRPM2-AS促进胆囊癌血管生成的作用机制的研究；PP4R1促进胆囊癌疾病进展的机制研究。仁济医院刘颖斌教授团队揭示了甘油三脂和酰基肉碱在胆囊癌中的恶性作用；陈涛教授团队研究发现DNMT3A与YAP/TAZ合作驱动胆囊癌转移的分子机制。以及我科团队对NONO促进 DLG1 外显子跳跃的分子机制探索。这些基础研究探讨了胆囊癌发展的分子机制，为未来的胆囊癌治疗的新靶点提供了重要的理论基础。同时，我们特别鸣谢上海市胆道疾病研究重点实验室邵荣教授、刘诗蕾博士、王化恺博士、赵成博士，对本次文献月评做出的贡献！
欢迎各位同道与我们积极交流探讨，共同推动胆囊癌研究和诊治的进步！
龚伟
1.外泌体运输的LncRNA TRPM2-AS 通过与PABPC1 相互作用激活 NOTCH1 信号通路促进胆囊癌的血管生成（IF: 37.3）
He Z, Zhong Y, Regmi P, et al. Exosomal long non-coding RNA TRPM2-AS promotes angiogenesis in gallbladder cancer through interacting with PABPC1 to activate NOTCH1 signaling pathway. Mol Cancer. 2024;23(1):65. Published 2024 Mar 27.
Methods: LncRNA sequencing was performed to identify the differentially expressed lncRNAs. RT-qPCR, western blot, FISH, and immunofluorescence were used to measure TRPM2-AS and NOTCH1 signaling pathway expression in vitro. Mouse xenograft and lung metastasis models were used to evaluate the biological function of TRPM2-AS during angiogenesis in vivo. EDU, transwell, and tube formation assays were used to detect the angiogenic ability of HUVECs. RIP, RAP, RNA pull-down, dual-luciferase reporter system, and mass spectrometry were used to confirm the interaction between TRPM2-AS, IGF2BP2, NUMB, and PABPC1.
Results: TRPM2-AS was upregulated in GBC tissues and was closely related to angiogenesis and poor prognosis in patients with GBC. The high expression level and stability of TRPM2-AS benefited from m6A modification, which is recognized by IGF2BP2. In terms of exerting pro-angiogenic effects, TRPM2-AS loaded with exosomes transported from GBC cells to HUVECs enhanced PABPC1-mediated NUMB expression inhibition, ultimately promoting the activation of the NOTCH1 signaling pathway. PABPC1 inhibited NUMB mRNA expression through interacting with AGO2 and promoted miR-31-5p and miR-146a-5p-mediated the degradation of NUMB mRNA. The NOTCH signaling pathway inhibitor DAPT inhibited GBC tumor angiogenesis, and TRPM2-AS knockdown enhanced this effect.
Conclusions: TRPM2-AS is a novel and promising biomarker for GBC angiogenesis that promotes angiogenesis by facilitating the activation of the NOTCH1 signaling pathway. Targeting TRPM2-AS opens further opportunities for future GBC treatments.
2.DNMT3A与YAP/TAZ合作驱动胆囊癌转移（IF: 15.1）
Xu S, Yuan Z, Jiang C, Chen W, Li Q, Chen T. DNMT3A Cooperates with YAP/TAZ to Drive Gallbladder Cancer Metastasis. Adv Sci (Weinh). Published online February 21, 2024.
3.NONO 通过与 IGF2BP3/RBM14 相互作用增强 DLG1 的致癌 RNA 剪接来促进胆囊癌细胞增殖（IF: 9.7）
Yang ZY, Zhao C, Liu SL, et al. NONO promotes gallbladder cancer cell proliferation by enhancing oncogenic RNA splicing of DLG1 through interaction with IGF2BP3/RBM14. Cancer Lett. 2024;587:216703. doi:10.1016/j.canlet.2024.216703
摘要：Gallbladder cancer (GBC) is a highly malignant and rapidly progressing tumor of the human biliary system, and there is an urgent need to develop new therapeutic targets and modalities. Non-POU domain-containing octamer-binding protein (NONO) is an RNA-binding protein involved in the regulation of transcription, mRNA splicing, and DNA repair. NONO expression is elevated in multiple tumors and can act as an oncogene to promote tumor progression. Here, we found that NONO was highly expressed in GBC and promoted tumor cells growth. The dysregulation of RNA splicing is a molecular feature of almost all tumor types. Accordingly, mRNA-seq and RIP-seq analysis showed that NONO promoted exon6 skipping in DLG1, forming two isomers (DLG1-FL and DLG1-S). Furthermore, lower Percent-Spliced-In (PSI) values of DLG1 were detected in tumor tissue relative to the paraneoplastic tissue, and were associated with poor patient prognosis. Moreover, DLG1-S and DLG1-FL act as tumor promoters and tumor suppressors, respectively, by regulating the YAP1/JUN pathway. N6-methyladenosine (m6A) is the most common and abundant RNA modification involved in alternative splicing processes. We identified an m6A reader, IGF2BP3, which synergizes with NONO to promote exon6 skipping in DLG1 in an m6A-dependent manner. Furthermore, IP/MS results showed that RBM14 was bound to NONO and interfered with NONO-mediated exon6 skipping of DLG1. In addition, IGF2BP3 disrupted the binding of RBM14 to NONO. Overall, our data elucidate the molecular mechanism by which NONO promotes DLG1 exon skipping, providing a basis for new therapeutic targets in GBC treatment.
4.PP4R1 通过促进 ERK1/2 介导的 PKM2 核易位来促进糖酵解和胆囊癌进展（IF: 9.7）
He Z, Zhong Y, Lv T, et al. PP4R1 promotes glycolysis and gallbladder cancer progression through facilitating ERK1/2 mediated PKM2 nuclear translocation. Cancer Lett. 2024;586:216677. doi:10.1016/j.canlet.2024.216677
摘要：Gallbladder cancer (GBC) is a common solid tumor of the biliary tract with a high mortality rate and limited curative benefits from surgical resection. Here, we aimed to elucidate the pathogenesis of GBC from the perspective of molecular mechanisms and determined that protein phosphatase 4 regulator subunit 1 (PP4R1) is overexpressed in GBC tissues and contributes to poor prognosis. Through a series of in vitro and in vivo experiments, we demonstrated that PP4R1 overexpression improved tumorigenesis in GBC cells. Further mechanistic exploration revealed that PP4R1 directly interacts with pyruvate kinase-M2 (PKM2), a key regulator of glycolysis. PP4R1 promotes the extracellular signal-related kinase 1 and 2 (ERK1/2)-mediated PKM2 nuclear translocation, thereby participating in the regulation of tumor glycolysis. Interestingly, we determined that PP4R1 strengthens the interaction between ERK1/2 and PKM2. Furthermore, PP4R1 enhanced the suppressive effects of the ERK inhibitor SCH772984 on GBC. In conclusion, our data showed that PP4R1 is a promising biomarker associated with GBC and confirmed that PP4R1 regulates PKM2-mediated tumor glycolysis, which provides a metabolic growth advantage to GBC cells, thereby promoting GBC tumor growth and metastasis.
通过经典的体外和体内实验，研究团队证实PP4R1的过表达显著提升了胆囊癌细胞的增殖、迁移和侵袭能力，以及这些细胞成瘤和促癌的能力。机制方面，研究揭示了PP4R1与PKM2之间存在直接相互作用，PP4R1能够通过激活ERK1/2的信号通路，进而促进PKM2的核内转运。核内的PKM2参与了调节GLUT1和LDHA在内的多种糖酵解相关基因的表达，从而加强了肿瘤细胞的糖酵解过程为其提供必要的能量支持，进而肿瘤细胞能够快速增殖和扩散。此外，文章还探索了PP4R1在药物治疗中的潜在应用价值，通过靶向PP4R1能够显著增强ERK抑制剂SCH772984对胆囊癌细胞的抑制效果，提供了将PP4R1作为新的治疗靶点的可能性。这项研究的成果不仅加深了我们对胆囊癌生物学机制的理解，还为其早期诊断、治疗策略的开发以及临床治疗方案的优化提供了新的思路和策略。这种对分子机制的深入剖析是开发针对性更强、副作用更小治疗药物的关键，未来有望改善胆囊癌患者的治疗效果和生存质量。
Yang Y, Li H, Liu K, et al. Acylcarnitines promote gallbladder cancer metastasis through lncBCL2L11-THOC5-JNK axis. J Transl Med. 2024;22(1):299. Published 2024 Mar 22. doi:10.1186/s12967-024-05091-0
摘要：Gallbladder cancer (GBC) is among the most common malignancies of biliary tract system due to its limited treatments. The immunotherapeutic targets for T cells are appealing, however, heterogeneity of T cells hinds its further development. We systematically construct T cell atlas by single-cell RNA sequencing; and utilized the identified gene signatures of high_CNV_T cells to predict molecular subtyping towards personalized therapeutic treatments for GBC. We identified 12 T cell subtypes, where exhausted CD8+ T cells, activated/exhausted CD8+ T cells, and regulatory T cells were predominant in tumors. There appeared to be an inverse relationship between Th17 and Treg populations with Th17 levels significantly reduced, whereas Tregs were concomitantly increased. Furthermore, we first established subtyping criterion to identify three subtypes of GBC based on their pro-tumorigenic microenvironments, e.g., the type 1 group shows more M2 macrophages infiltration, while the type 2 group is infiltrated by highly exhausted CD8+ T cells, B cells and Tregs with suppressive activities. Our study provides valuable insights into T cell heterogeneity and suggests that molecular subtyping based on T cells might provide a potential immunotherapeutic strategy to improve GBC treatment.
