PO.CL01.13 · 临床研究
胆囊癌从前驱阶段到未分化阶段的空间分辨转录组学特征
Spatially resolved transcriptomic signatures of gallbladder carcinogenesis from precursor to undifferentiated stages
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：胆道上皮内瘤变（BilIN）被认为是胆囊癌的前驱病变。然而，癌变序贯过程背后的转录组学改变仍知之甚少。此外，胆囊癌的分化状态在患者之间各不相同，一些肿瘤在同一肿瘤内同时表现出分化和未分化成分。为阐明这一组织学谱系中的关键转录组变化，我们对包含正常上皮、BilIN、分化型癌和未分化型癌共存区域的胆囊肿瘤样本进行了空间转录组分析，以了解个体患者内不同病变之间的分子差异。
方法：从13例患者中选取了共59个感兴趣区域（ROI），包括正常（n=11）、异型增生（n=14）、分化型癌（n=22）、未分化型癌（n=8）和转移病变（n=4）。使用GeoMx数字空间分析（GeoMx DSP）结合EpCAM染色进行空间转录组分析。每个ROI的最大尺寸为750×625 µm，且每个ROI至少包含200个细胞。从GeoMx DSP衍生的表达谱中，通过A-正交非负矩阵分解（AONMF）对基因表达动态进行模式化，并通过加权基因共表达网络分析（WGCNA）推断上皮-基质串扰。
结果：通路富集分析显示，胆汁酸代谢和脂肪酸代谢在正常谱系中最高，并随组织学恶化而逐渐减弱。从分化谱系起，细胞增殖程序上调，而未分化谱系以EMT特征为特点。上皮-基质相互作用分析显示，上皮区域存在TNC-ITGB1轴，该轴在未分化型癌内激活基质细胞中的TGF-beta信号传导。携带未分化型癌的患者在分化区域内表现出独特的转录模式，与未进展为未分化型癌的患者相比，HDGFL3增加而MMP1表达降低，提示HDGFL3上调可能代表去分化之前的早期分子线索。
结论：本研究提供了对胆囊肿瘤发生序贯癌变过程的空间转录组学见解。EMT通路在未分化谱系中升高，与TNC-ITGB1轴相关，且HDGFL3在携带未分化成分患者的分化区域中上调。因此，HDGFL3上调可能作为从BilIN进展为未分化型胆囊癌的潜在预测性生物标志物。
查看英文原文 English abstract
Background: Biliary intraepithelial neoplasia (BilIN) is recognized as a precursor lesion of gallbladder carcinoma. However, the transcriptomic alterations underlying the sequential process of carcinogenesis remain poorly understood. Moreover, the differentiation status of gallbladder carcinoma varies among patients, with some tumors exhibiting both differentiated and undifferentiated components within the same tumor. To elucidate key transcriptomic changes across this histologic spectrum, we performed spatial transcriptomic analysis on gallbladder tumor samples containing coexisting areas of normal epithelium, BilIN, differentiated carcinoma, and undifferentiated carcinoma to understand the molecular differences between lesions within individual patients.
Methods: A total of 59 regions of interest (ROIs) were selected from 13 patients, including normal (n = 11), dysplasia (n = 14), differentiated carcinoma (n = 22), undifferentiated carcinoma (n = 8), and metastatic lesions (n = 4). Spatial transcriptome profiling was performed using GeoMx Digital Spatial Profiling (GeoMx DSP) with EpCAM staining. The maximum size of each ROI was 750 × 625 µm, and each ROI included at least 200 cells. From GeoMx DSP-derived expression profiles, gene-expression dynamics were patterned via A-orthogonal nonnegative matrix factorization (AONMF), and epithelial-stromal crosstalk was inferred via weighted gene co-expression network analysis (WGCNA).
Results: Pathway enrichment analysis revealed bile acid metabolism and fatty acid metabolism were highest in normal spectrum and progressively diminished with histological worsening. From the differentiated spectrum, cell proliferation programs were upregulated, and the undifferentiated spectrum was characterized by EMT signature. Epithelial-stromal interaction analysis showed a TNC-ITGB1 axis in epithelial regions that activates TGF-beta signaling in stromal cells within undifferentiated carcinoma. Patients harboring undifferentiated carcinoma exhibited distinct transcriptional patterns within the differentiated regions, showing increased HDGFL3 and decreased MMP1 expression compared with patients without progression to undifferentiated carcinoma, suggesting that HDGFL3 upregulation may represent an early molecular cue preceding dedifferentiation.
Conclusion: This study provides spatial transcriptomic insights into the sequential carcinogenic process of gallbladder tumorigenesis. EMT pathway is elevated in the undifferentiated spectrum in association with the TNC-ITGB1 axis, and HDGFL3 is upregulated in differentiated regions of patients harboring undifferentiated components. Thus, upregulation of HDGFL3 may serve as potential predictive biomarkers for the progression from BilIN to undifferentiated gallbladder carcinoma.
利益披露 Disclosure
Y. Kim, None..
Y. Lee, None..
H. Oh, None..
Y. Sung, None..
Y. Lee, None..
Y. Kim, None..
I. Park, None..
J. Bhin, None..
S. Shin, None.