胆囊癌文献月评（202405期）

健康健康 2024-06-29 10:00 上海

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呈现了多个领域的新进展，涵盖了TOPAZ-1临床研究的报告结果、Olfactomedin 4在胆囊癌发病机制中的潜在作用、术中冰冻切片诊断方法、TAT在胆囊癌肝转移过程中的生物学功能以及禁食诱导的RNF152提高胆囊癌细胞对吉西他滨敏感性的分子机制等方面。这些研究在不同层面上揭示了胆囊癌治疗的新可能性，进一步推动了胆囊癌临床和基础研究的发展。因此，我们特别鸣谢上海交通大学附属新华医院肿瘤科周晴教授，上海交通大学附属新华医院病理科管雯斌教授，上海交通大学附属新华医院普外科李冬冬医师，以及上海市胆道疾病研究重点实验室李春杰研究员、吴自友博士、张健硕士做出的贡献！

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.Durvalumab 联合吉西他滨和顺铂治疗晚期胆道癌（TOPAZ-1）：一项随机、双盲、安慰剂对照的3期试验的患者报告结果（IF:51.1）

Burris HA 3rd, Okusaka T, Vogel A, et al. Durvalumab plus gemcitabine and cisplatin in advanced biliary tract cancer (TOPAZ-1): patient-reported outcomes from a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet Oncol. 2024;25(5):626-635.

摘要：Background: In the ongoing, randomised, double-blind phase 3 TOPAZ-1 study, durvalumab, a PD-L1 inhibitor, plus gemcitabine and cisplatin was associated with significant improvements in overall survival compared with placebo, gemcitabine, and cisplatin in people with advanced biliary tract cancer at the pre-planned intermin analysis. In this paper, we present patient-reported outcomes from TOPAZ-1.Methods: In TOPAZ-1 (NCT03875235), participants aged 18 years or older with previously untreated, unresectable, locally advanced, or metastatic biliary tract cancer with an Eastern Cooperative Oncology Group performance status of 0 or 1 and one or more measurable lesions per Response Evaluation Criteria in Solid Tumors (RECIST; version 1.1) were randomly assigned (1:1) to the durvalumab group or the placebo group using a computer-generated randomisation scheme. Participants received 1500 mg durvalumab or matched placebo intravenously every 3 weeks (on day 1 of the cycle) for up to eight cycles in combination with 1000 mg/m2 gemcitabine and 25 mg/m2 cisplatin intravenously on days 1 and 8 every 3 weeks for up to eight cycles. Thereafter, participants received either durvalumab (1500 mg) or placebo monotherapy intravenously every 4 weeks until disease progression or other discontinuation criteria were met. Randomisation was stratified by disease status (initially unresectable vs recurrent) and primary tumour location (intrahepatic cholangiocarcinoma vs extrahepatic cholangiocarcinoma vs gallbladder cancer). Patient-reported outcomes were assessed as a secondary outcome in all participants who completed the European Organisation for Research and Treatment of Cancer's 30-item Quality of Life of Cancer Patients questionnaire (QLQ-C30) and the 21-item Cholangiocarcinoma and Gallbladder Cancer Quality of Life Module (QLQ-BIL21). We calculated time to deterioration-ie, time from randomisation to an absolute decrease of at least 10 points in a patient-reported outcome that was confirmed at a subsequent visit or the date of death (by any cause) in the absence of deterioration-and adjusted mean change from baseline in patient-reported outcomes.Findings: Between April 16, 2019, and Dec 11, 2020, 685 participants were enrolled and randomly assigned, 341 to the durvalumab group and 344 to the placebo group. Overall, 345 (50%) of participants were male and 340 (50%) were female. Data for the QLQ-C30 were available for 318 participants in the durvalumab group and 328 in the placebo group (median follow-up 9·9 months [IQR 6·7 to 14·1]). Data for the QLQ-BIL21 were available for 305 participants in the durvalumab group and 322 in the placebo group (median follow-up 10·2 months [IQR 6·7 to 14·3]). The proportions of participants in both groups who completed questionnaires were high and baseline scores were mostly similar across treatment groups. For global health status or quality of life, functioning, and symptoms, we noted no difference in time to deterioration or adjusted mean changes from baseline were observed between groups. Median time to deterioration of global health status or quality of life was 7·4 months (95% CI 5·6 to 8·9) in the durvalumab group and 6·7 months (5·6 to 7·9) in the placebo group (hazard ratio 0·87 [95% CI 0·69 to 1·12]). The adjusted mean change from baseline was 1·23 (95% CI -0·71 to 3·16) in the durvalumab group and 0·35 (-1·63 to 2·32) in the placebo group.Interpretation: The addition of durvalumab to gemcitabine and cisplatin did not have a detrimental effect on patient-reported outcomes. These results suggest that durvalumab, gemcitabine, and cisplatin is a tolerable treatment regimen in patients with advanced biliary tract cancer.

周晴教授（上海交通大学附属新华医院肿瘤科）：

这是一项关于度伐利尤单抗联合吉西他滨和顺铂治疗晚期胆道癌的随机、双盲、安慰剂对照的三期研究(TOPAZ-1)。与安慰剂相比，度伐利尤单抗（PD-L1抑制剂）加上吉西他滨和顺铂组总生存期得到显著改善。

在TOPAZ-1 (NCT03875235)中，入组年龄在18岁及以上，患有既往未经治疗的、不可切除的、局部进展或转移性胆道癌。从2019年4月16日至2020年12月11日期间，685名参与者被招募并随机分配(1:1)，341名被分配到度伐利尤单抗组，安慰剂组344例。每3周的第1天分别接受1500 mg 度伐利尤单抗或安慰剂静脉注射，联合1000mg /m²吉西他滨和25 mg/m²顺铂静脉滴注（每3周的第1天和第8天，最多8个周期）。之后每4周静脉注射度伐利尤单抗(1500mg)或安慰剂单药治疗，直至病情进展或符合其他停药标准。按疾病状态随机化分层：最初不可切除vs复发；原发肿瘤位置(肝内胆管癌vs肝外胆管癌，胆管癌vs胆囊癌)。入组者完成了欧洲癌症研究和治疗组织的30项癌症患者的生活质量问卷(QLQ-C30)和21项胆管癌和胆囊癌患者生活质量问卷(QLQ-BIL21)。度伐利尤单抗组318名入组者和安慰剂组328名入组者的QLQ-C30数据被采用，中位随访时间为9.9个月[IQR为6.7 ~ 14.1]，度伐利尤单抗组305例和安慰剂组322例的QLQ-BIL21数据被采用，中位随访10.2个月[IQR 6.7～14.3]。两组入组者完成问卷的比例都很高，各治疗组的基线得分情况相似。度伐利尤单抗组总体健康状况或生活质量恶化的中位时间为7.4个月(95% CI 5.6 ~ 8.9)，安慰剂组为6.7个月(5.6 ~ 7.9)(风险比0.87 [95% CI 0.69 ~ 1.12])。与基线相比，度伐利尤单抗组调整后平均变化值为1.23 (95% CI - 0.71至3.16)，安慰剂组为0.35(- 1.63至2.32)。统计出的恶化时间至少有10个点的绝对下降，在之后的随访或是非恶化的死亡时间（任何原因）得到证实。

在吉西他滨和顺铂的基础上加用度伐利尤单抗对患者未增加明显毒副作用，是晚期胆道癌患者可以耐受的治疗方案。

2.术中冰冻切片分析指导的疑似胆囊癌的单期管理：一项回顾性队列研究（IF:15.3）

Banh S, Fehervari M, Flod S, et al. Single stage management of suspected gallbladder cancer guided by intraoperative frozen section analysis: A retrospective cohort study. Int J Surg. Published online May 3, 2024.

摘要：Background: The management of radiologically suspected gallbladder cancers (GBC) that lack definitive radiological features usually involves performing a first-stage routine laparoscopic cholecystectomy, followed by an open second-stage liver resection (segments IVB and V) and hilar lymphadenectomy (extended cholecystectomy) if subsequent formal histology confirms a malignancy. Performing a cholecystectomy with an intraoperative frozen section to guide the need for conversion to an extended cholecystectomy as a single-stage procedure has multiple benefits compared to a two-stage approach. However, the safety and efficacy of this approach have not yet been evaluated in a tertiary setting.Methods: A retrospective cohort study was performed using a database of all consecutive patients with suspected GBC who had been referred to our tertiary unit. Following routine cholecystectomy, depending on the operative findings, the gallbladder specimen was removed and sent for frozen-section analysis. If malignancy was confirmed, the depth of tumour invasion was evaluated, followed by simultaneous extended cholecystectomy, when appropriate. The sensitivity and specificity of frozen section analysis for the diagnosis of GBC were measured using formal histopathology as a reference standard.Results: A total of 37 consecutive cholecystectomies were performed. In nine cases, GBC was confirmed by intraoperative frozen section analysis, three of which had standard cholecystectomy only as their frozen section showed adenocarcinoma to be T1a or below (n=2) or were undetermined (n=1). In the remaining six cases, malignant invasion beyond the muscularis propria (T1b or above) was confirmed; thus, a synchronous extended cholecystectomy was performed. The sensitivity (95% CI 66.4%-100%) and specificity (95% CI 87.7%-100%) for identifying GBC using frozen section analysis were both 100%. The net cost of the single-stage pathway in comparison to the two-stage pathway resulted in overall savings of £3894.Conclusion: Intraoperative frozen section analysis is a reliable tool for guiding the use of a safe, single-stage approach for the management of GBC in radiologically equivocal cases. In addition to its lower costs compared to a conventional two-stage procedure, intraoperative analysis also affords the benefit of a single hospital admission and single administration of general anaesthesia, thus greatly enhancing the patient's experience and relieving the burden on waiting lists.

管雯斌教授（上海交通大学附属新华医院病理科）：

胆囊癌是较常见的胆道恶性肿瘤，但由于早期症状隐匿，发现时往往已是晚期。本研究为单中心研究，通过回顾37例胆囊术中冰冻检查诊断，证实了对于怀疑胆囊癌的标本进行术中冰冻检查诊断的可行性。术中冰冻诊断除了可以明确有无恶性病变外，还可以明确浸润深度，为确定手术范围提供依据；且冰冻诊断与术后石蜡诊断有较高的一致性。该研究认为，对于胆囊术中冰冻检查诊断最主要有以下优点：1. 减少二次手术概率，提供了单次住院及单次麻醉的优势，改善了患者的体验。2. 术中冰冻检查并未明显增加术中等待时间及手术成本。因此，该研究认为在胆囊癌术前诊断不明朗的情况下，术中冰冻检查可靠地指导单阶段手术且具有较高的成本效益。

作者也讨论了其他术前诊断的可行性，尽管超声引导下细针穿刺具有较高的诊断敏感性，但有造成腹膜播散的风险，且无法评估肿瘤浸润深度。故在该研究中，对于临床怀疑胆囊癌的患者，影像怀疑或不确定，伴有胆石症、高龄等高危因素，经MDT讨论后，会进行术中冰冻检查。研究结果明确了术中冰冻检查在胆囊癌诊断中具有较高的敏感性和特异性。

但作者也提到该研究中具有可疑放射学特征的病例相对较少。若将该研究扩展为多中心前瞻性随机对照试验，并引入远程病理诊断，可突破该研究的局限性。

在日常工作中，胆囊术中冰冻检查缺失可以协助外科医师决定手术范围，确定切缘情况，且大部分病例术中冰冻病理诊断并不困难。而胆囊癌发病较为隐匿，哪些患者需术中冰冻检查，是否适当放宽胆囊术中冰冻检查指征，仍有待进一步研究、讨论。

3.Durvalumab 联合吉西他滨和顺铂与吉西他滨和顺铂治疗BTC的比较：一项基于真实世界的多中心回顾性研究（IF:5.4）

M, Masi G, Lonardi S, et al. Durvalumab Plus Gemcitabine and Cisplatin Versus Gemcitabine and Cisplatin in Biliary Tract Cancer: a Real-World Retrospective, Multicenter Study. Target Oncol. 2024;19(3):359-370.

摘要：Background: The TOPAZ-1 phase III trial reported a survival benefit with the anti-programmed cell death ligand 1 (anti-PD-L1) durvalumab in combination with gemcitabine and cisplatin in patients with advanced biliary tract cancer (BTC).Objective: The present study investigated for the first time the impact on survival of adding durvalumab to cisplatin/gemcitabine compared with cisplatin/gemcitabine in a real-world setting.Patients and methods: The analyzed population included patients with unresectable, locally advanced, or metastatic BTC treated with durvalumab in combination with cisplatin/gemcitabine or with cisplatin/gemcitabine alone. The impact of adding durvalumab to chemotherapy in terms of overall survival (OS) and progression free survival (PFS) was investigated with univariate and multivariate analysis.Results: Overall, 563 patients were included in the analysis: 213 received cisplatin/gemcitabine alone, 350 received cisplatin/gemcitabine plus durvalumab. At the univariate analysis, the addition of durvalumab was found to have an impact on survival, with a median OS of 14.8 months versus 11.2 months [hazard ratio (HR) 0.63, 95% confidence interval (CI) 0.50-0.80, p = 0.0002] in patients who received cisplatin/gemcitabine plus durvalumab compared to those who received cisplatin/gemcitabine alone. At the univariate analysis for PFS, the addition of durvalumab to cisplatin/gemcitabine demonstrated a survival impact, with a median PFS of 8.3 months and 6.0 months (HR 0.57, 95% CI 0.47-0.70, p < 0.0001) in patients who received cisplatin/gemcitabine plus durvalumab and cisplatin/gemcitabine alone, respectively. The multivariate analysis confirmed that adding durvalumab to cisplatin/gemcitabine is an independent prognostic factor for OS and PFS, with patients > 70 years old and those affected by locally advanced disease experiencing the highest survival benefit. Finally, an exploratory analysis of prognostic factors was performed in the cohort of patients who received durvalumab: neutrophil-lymphocyte ratio (NLR) and disease stage were to be independent prognostic factors in terms of OS. The interaction test highlighted NLR ≤ 3, Eastern Cooperative Oncology Group Performance Status (ECOG PS) = 0, and locally advanced disease as positive predictive factors for OS on cisplatin/gemcitabine plus durvalumab.Conclusion: In line with the results of the TOPAZ-1 trial, adding durvalumab to cisplatin/gemcitabine has been confirmed to confer a survival benefit in terms of OS and PFS in a real-world setting of patients with advanced BTC.

李冬冬医师（上海交通大学附属新华医院普外科）：

众所周知，胆道恶性肿瘤（BTC）是一类异质性强、预后较差、治疗选择相对有限的肿瘤。当前，晚期BTC仍然是肿瘤学领域的一大临床挑战，其主要治疗手段是以全身系统性化疗为主的综合治疗。前序已报道，基于TOPAZ-1 试验的III 期研究结果，Durvalumab联合吉西他滨和顺铂方案已被列为先前未经治疗的不可切除或转移性BTC患者的新一线治疗方案，并于去年初步发布了这一治疗组合方案在现实世界中的安全性和有效性验证研究的结果。在此基础上，本项研究进一步回顾性地比较了两组患者——一组接受之前的标准治疗（顺铂和吉西他滨），另一组接受新的治疗组合方案（Durvalumab联合顺铂和吉西他滨），其目的是评估在化疗中加入Durvalumab对生存的影响。此外，对Durvalumab联合顺铂/吉西他滨反应的患者预后和预测因素进行了探索性分析。

研究共纳入了 563 名患者，其中213 名患者仅接受顺铂和吉西他滨治疗，另外350 名患者接受Durvalumab联合顺铂和吉西他滨治疗。单因素分析发现化疗添加Durvalumab对生存有影响。两组相比，接受Durvalumab联合顺铂和吉西他滨治疗的患者中位OS为 14.8 个月，而仅接受顺铂/吉西他滨治疗的患者中位OS为11.2 个月 (p = 0.0002)；在 PFS 方面，接受Durvalumab联合顺铂和吉西他滨治疗与仅接受顺铂和吉西他滨治疗的患者的中位 PFS 分别为 8.3 个月和 6.0 个月（p < 0.0001）。多因素分析证实，Durvalumab联合顺铂和吉西他滨是 OS 和 PFS 的独立预后因素。同时，在接受度伐单抗治疗的患者队列中进行了预后因素的探索性分析：中性粒细胞-淋巴细胞比率 (NLR) 和疾病分期是 OS 的独立预后因素。

该项研究结果进一步证实Durvalumab联合顺铂和吉西他滨可在现实世界中为晚期 BTC 患者的 OS 和 PFS 带来生存获益，也为晚期BTC患者的治疗带来了好声音。但与此同时，我们也应当注意到回顾性研究的选择偏差可能性；多中心或机构之间肿瘤评估方式或时间节点的细微差异性；以及相对更为重要的随访时间的长短性。我们期待研究在未来更长时间的随访后进行两组患者生存结果的更新，或许将更有助于印证目前的结果。

4.禁食诱导的RNF152通过抑制mTORC1调控的糖酵解增强胆囊癌细胞对吉西他滨化疗药物的敏感性（IF: 5.8）

Tao Y, Gong Z, Shen S, et al. Fasting-induced RNF152 resensitizes gallbladder cancer cells to gemcitabine by inhibiting mTORC1-mediated glycolysis. iScience. 2024;27(5):109659. Published 2024 Apr 8.

摘要：Abnormal mTORC1 activation by the lysosomal Ragulator complex has been implicated in cancer and glycolytic metabolism associated with drug resistance. Fasting upregulates RNF152 and mediates the metabolic status of cells. We report that RNF152 regulates mTORC1 signaling by targeting a Ragulator subunit, p18, and attenuates gemcitabine resistance in gallbladder cancer (GBC). We detected levels of RNF152 and p18 in tissues and undertook mechanistic studies using activators, inhibitors, and lentivirus transfections. RNF152 levels were significantly lower in GBC than in adjacent non-cancer tissues. Fasting impairs glycolysis, induces gemcitabine sensitivity, and upregulates RNF152 expression. RNF152 overexpression increases the sensitivity of GBC cells to gemcitabine, whereas silencing RNF152 has the opposite effect. Fasting-induced RNF152 ubiquitinates p18, resulting in proteasomal degradation. RNF152 deficiency increases the lysosomal localization of p18 and increases mTORC1 activity, to promote glycolysis and decrease gemcitabine sensitivity. RNF152 suppresses mTORC1 activity to inhibit glycolysis and enhance gemcitabine sensitivity in GBC.

李春杰研究员（上海市胆道疾病研究重点实验室）：

禁食（Fasting），作为一种有效的饮食干预手段，已被证实可以通过调节肿瘤细胞的能量代谢来控制肿瘤细胞生长，提高肿瘤细胞放化疗的敏感性，但禁食对胆囊癌代谢变化的影响以及其对吉西他滨化疗敏感性的研究很少。复旦中山医院胆道外科索涛教授团队于2024年5月在《Cell》子刊《iScience》上发表题为“Fasting-induced RNF152 resensitizes gallbladder cancer cells to gemcitabine by inhibiting mTORC1-mediated glycolysis”的研究论文。该项研究发现RNF152是禁食的关键下游靶标，禁食能够上调RNF152的表达，下调p18抑制mTORC1介导的糖酵解通路，该研究阐明了禁食能够增强胆囊癌细胞对吉西他滨敏感性的详细分子机制，为禁食在干预治疗胆囊癌方面提供了新的思路。RING家族E3泛素连接酶通过介导底物蛋白的降解，参与调控肿瘤的发生、发展和耐药性。研究团队首先发现禁食确实可以抑制糖酵解并促进GBC细胞对GEM的敏感性。RNF152基因的表达下调与胆囊癌恶性进展和不良预后相关。RNF152在胆囊癌中起着至关重要的肿瘤抑制作用。其次发现，禁食会上调RNF152的表达，体内外实验证实，禁食和过表达RNF152可以显著抑制胆囊癌细胞的生长，且联合抑制效果更明显。此外，团队发现体内外敲低RNF152均显著降低了胆囊癌化疗药物吉西他滨的敏感性，而禁食和过表达RNF152可以通过抑制mTORC1信号通路抑制胆囊癌糖酵解过程，增强胆囊癌细胞对吉西他滨的敏感性。机制上，作者发现RNF152可以通过泛素化作用破坏p18复合体的稳定性，负向调控mTORC1通路。综上，该研究阐明了禁食诱导的RNF152提高胆囊癌细胞对吉西他滨敏感性的分子机制，为禁食在辅助干预治疗胆囊癌方面提供了新的依据。展更多的高质量前瞻性临床研究，解决以上不足之处，为更多的 GBC 患者带来生存优势。

5.单细胞综合分析破译胆囊癌发病机制中的微环境动态和免疫调节因子嗅觉调节素 4（IF：24.5）

He H, Chen S, Yu Y, et al. Comprehensive single-cell analysis deciphered microenvironmental dynamics and immune regulator olfactomedin 4 in pathogenesis of gallbladder cancer. Gut. Published online May 6, 2024.

摘要：Objective: Elucidating complex ecosystems and molecular features of gallbladder cancer (GBC) and benign gallbladder diseases is pivotal to proactive cancer prevention and optimal therapeutic intervention.Design: We performed single-cell transcriptome analysis on 230 737 cells from 15 GBCs, 4 cholecystitis samples, 3 gallbladder polyps, 5 gallbladder adenomas and 16 adjacent normal tissues. Findings were validated through large-scale histological assays, digital spatial profiler multiplexed immunofluorescence (Gemox), etc. Further molecular mechanism was demonstrated with in vitro and in vivo studies.Results: The cell atlas unveiled an altered immune landscape across different pathological states of gallbladder diseases. GBC featured a more suppressive immune microenvironment with distinct T-cell proliferation patterns and macrophage attributions in different GBC subtypes. Notably, mutual exclusivity between stromal and immune cells was identified and remarkable stromal ecosystem (SC) heterogeneity during GBC progression was unveiled. Specifically, SC1 demonstrated active interaction between Fibro-iCAF and Endo-Tip cells, correlating with poor prognosis. Moreover, epithelium genetic variations within adenocarcinoma (AC) indicated an evolutionary similarity between adenoma and AC. Importantly, our study identified elevated olfactomedin 4 (OLFM4) in epithelial cells as a central player in GBC progression. OLFM4 was related to T-cell malfunction and tumour-associated macrophage infiltration, leading to a worse prognosis in GBC. Further investigations revealed that OLFM4 upregulated programmed death-ligand 1 (PD-L1) expression through the MAPK-AP1 axis, facilitating tumour cell immune evasion.Conclusion: These findings offer a valuable resource for understanding the pathogenesis of gallbladder diseases and indicate OLFM4 as a potential biomarker and therapeutic target for GBC.

吴自友博士（上海市胆道疾病研究重点实验室）：

这是一项由王红阳院士、文文教授团队在《Gut》发表的研究，针对43例样本（包括4例胆囊炎、3例胆固醇性息肉、4例胆囊腺瘤、15例胆囊癌及癌旁组织）进行了单细胞测序。将230,737个细胞分为7种主要的细胞类型并建立了包含69个亚群的细胞图谱，对比分析了各组的独特模式。研究发现GBC具有丰富的免疫抑制免疫细胞（如CD4-Treg和CD8-Tex）和基质细胞（Fibro-iCAF），这与它们在肿瘤发生中的作用一致。通过整合全基因组、二代转录组及GeoMx空间组学等多组学数据，并结合传统生物学实验手段，全面揭示了胆囊癌及其前驱疾病状态的细胞景观与分子特征，解析了疾病演化过程中的微环境异质性。研究发现胆囊癌显示出较强的免疫抑制性微环境，不同病理亚型的胆囊癌在T细胞增殖模式、T细胞受体（TCR）克隆扩增特征及巨噬细胞特性上存在差异，为针对关键免疫细胞亚群的治疗策略提供了理论基础。基质细胞分析揭示了基质-免疫细胞间显著的互斥性，展示了3类胆囊疾病进程中的复杂基质生态系统（Stromal Ecosystem，SC），其中SC1和SC3主要出现在胆囊癌样本中，以Fibro-iCAF和Endo-Tip亚群间的活跃交互作用为特征的SC1与预后不良相关。此外，该研究发现，嗅素蛋白4（Olfactomedin 4，OLFM4）在胆囊疾病演进过程中逐渐升高，并可在外周血中检测到，与胆囊癌病人预后不良密切相关。进一步研究发现，OLFM4与T细胞功能障碍和肿瘤相关巨噬细胞浸润密切相关，可通过MAPK- AP1信号轴调控胆囊癌细胞表面程序性死亡配体1（PD-L1）的表达，进而在促肿瘤免疫逃逸中发挥关键作用。

总体上，此项通过全基因组测序（WGS）、单细胞测序（scRNA-seq）和数字空间表达谱（GeoMx DSP）揭示了Olfactomedin 4在胆囊癌发病机制中的潜在作用，并阐明胆囊癌和良性胆囊疾病的复杂生态系统和分子特征是积极预防癌症和优化治疗的关键。

6.TRIM21 介导的 TAT 泛素化抑制胆囊癌的肝转移（IF：9.7）

Wu Z, Zhang J, Jia Z, et al. TRIM21-mediated ubiquitylation of TAT suppresses liver metastasis in gallbladder cancer. Cancer Lett. 2024;592:216923.

摘要：Liver metastasis is common in patients with gallbladder cancer (GBC), imposing a significant challenge in clinical management and serving as a poor prognostic indicator. However, the mechanisms underlying liver metastasis remain largely unknown. Here, we report a crucial role of tyrosine aminotransferase (TAT) in liver metastasis of GBC. TAT is frequently up-regulated in GBC tissues. Increased TAT expression is associated with frequent liver metastasis and poor prognosis of GBC patients. Overexpression of TAT promotes GBC cell migration and invasion in vitro, as well as liver metastasis in vivo. TAT knockdown has the opposite effects. Intriguingly, TAT promotes liver metastasis of GBC by potentiating cardiolipin-dependent mitophagy. Mechanistically, TAT directly binds to cardiolipin and leads to cardiolipin externalization and subsequent mitophagy. Moreover, TRIM21 (Tripartite Motif Containing 21), an E3 ubiquitin ligase, interacts with TAT. The histine residues 336 and 338 at TRIM21 are essential for this binding. TRIM21 preferentially adds the lysine 63 (K63)-linked ubiquitin chains on TAT principally at K136. TRIM21-mediated TAT ubiquitination impairs its dimerization and mitochondrial location, subsequently inhibiting tumor invasion and migration of GBC cells. Therefore, our study identifies TAT as a novel driver of GBC liver metastasis, emphasizing its potential as a therapeutic target.

张健硕士（上海市胆道疾病研究重点实验室）：

肝脏是胆囊癌发生远处转移最常见的部位，胆囊癌肝转移也是导致晚期胆囊癌疗效不佳和预后较差的重要原因。目前针对胆囊癌肝转移的放化疗获益有限，容易出现耐药，且缺乏研究证实临床疗效的靶向或免疫药物。

龚伟教授团队针对胆囊癌肝转移的机制进行了深入的研究，发现TAT（酪氨酸氨基转移酶）在胆囊癌肝转移中高表达，且与不良预后相关。TAT过表达可以促进胆囊癌细胞的侵袭、迁移能力和体内肝转移能力。研究团队进一步通过代谢组学和转录组学发现TAT并非通过经典的代谢依赖途径促进胆囊癌肝转移，而是通过促进心磷脂外化介导的线粒体自噬，进而增强胆囊癌细胞的肝转移能力。以往文献曾报道代谢相关酶具有许多潜在的非经典/非代谢功能，可以调控肿瘤微环境的重塑、DNA损伤修复、细胞增殖、存活、凋亡以及细胞周期等多种复杂的细胞活动。代谢酶不仅直接调节细胞增殖和存活的关键信号通路，还通过调节细胞自噬、线粒体功能以及氧化还原稳态来影响细胞的存活和凋亡。

此外，研究团队发现三基序蛋白21（TRIM21）是调控TAT的关键E3连接酶，在H336/338位点与TAT的K136位点相互作用，并通过K63泛素链对TAT进行多聚泛素化修饰。TRIM21介导的泛素化修饰抑制TAT二聚化和其在线粒体中的表达水平，并最终影响胆囊癌细胞的侵袭、迁移和体内肝转移能力。

本研究深入探讨了TAT在胆囊癌肝转移过程中的生物学功能，不仅有助于剖析胆囊癌肝转移的分子机制，还能为开发特异性分子诊断和靶向药物治疗提供重要理论依据，有助于改善胆囊癌肝转移患者预后极差的困境。

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上海交通大学附属新华医院普外科

上海交通大学医学院附属新华医院普外科创建于1958年，学科医、教、研水平居国内领先，以消化道肿瘤、器官移植、甲乳外科、微创外科为特色。普外科是卫生部国家临床重点专科，是卫生部首批批准的肝移植定点医院。2016年成立上海市胆道疾病研究中心。