PO.MCB08.04 · 分子与细胞生物学
利用基因工程小鼠模型对表现为黏液腺癌的结肠肿瘤进行空间转录组学分析
Spatial transcriptomics of colonic neoplasia presenting mucinous adenocarcinoma with genetically-engineered mouse model
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：我们此前建立了一种基因工程小鼠模型，其中Apc和Tgfbr2在结肠上皮中被特异性敲除（CDX2P-G19Cre; Apc flox/+; Tgfbr2 flox/flox）。在该模型中，40%的小鼠在结肠中发生黏液腺癌（MAC），而25%的小鼠在相同基因型下表现为非黏液性高级别异型增生。黏液性肿瘤侵犯超出黏膜肌层，并在深部浸润病灶中观察到beta-catenin的核定位。为阐明黏液性结直肠癌发生的机制，我们此前进行了比较MAC与非黏液性异型增生的转录组分析。在本研究中，我们进行了空间转录组分析，以表征MAC组织内基因表达、细胞组成及细胞间相互作用的空间分布。方法：从CDX2P-G19Cre; Apc flox/+; Tgfbr2 flox/flox小鼠中获取经H&E染色诊断为MAC的肿瘤。将FFPE组织块切片以制备文库，并使用NovaSeq平台进行测序。数据采用Space Ranger软件分析，并在Loupe Browser（10x Genomics）中可视化。结果：空间转录组谱分析揭示了同一MAC病灶内异质性的肿瘤微环境。在最深部浸润病灶中，鉴定出SPP1（分泌型磷蛋白1）阳性巨噬细胞浸润增加，提示存在与浸润相关的局部免疫调节生态位。结论：空间转录组分析显示了以独特细胞组成和基因表达谱为特征的不同瘤内区域。这些发现为黏液性结直肠癌的空间异质性和进展机制提供了新的见解，有望为未来的治疗策略提供信息。
查看英文原文 English abstract
Background: We previously established a genetically engineered mouse model in which Apc and Tgfbr2 are specifically knocked out in the colonic epithelium (CDX2P-G19Cre; Apc flox/+ ; Tgfbr2 flox/flox ). In this model, 40% of mice developed mucinous adenocarcinoma (MAC) in the colon, while 25% exhibited non-mucinous high-grade dysplasia with the same genotype. The mucinous tumors invaded beyond the muscularis mucosae, and nuclear localization of beta-catenin was observed in the deep invasive lesion. To elucidate the mechanisms underlying the development of mucinous colorectal carcinoma, we previously performed transcriptomic analysis comparing MAC and non-mucinous dysplasia. In the present study, we conducted spatial transcriptomic analysis to characterize the spatial distribution of gene expression, cellular composition, and cell-cell interactions within MAC tissues. Methods: Tumors diagnosed as MAC by H&E staining were obtained from CDX2P-G19Cre; Apc flox/+ ; Tgfbr2 flox/flox mice. FFPE tissue blocks were sectioned to prepare libraries and sequencing was performed using the NovaSeq platform. Data were analyzed using Space Ranger software and visualized in Loupe Browser (10x Genomics). Results: Spatial transcriptomic profiling revealed heterogeneous tumor microenvironments within the same MAC lesion. In the deepest invasive lesion, increased infiltration of SPP1 (secreted phosphoprotein 1)-positive macrophages was identified, suggesting the presence of a localized immunomodulatory niche associated with invasion. Conclusions: Spatial transcriptomic analysis demonstrated distinct intratumoral regions characterized by unique cellular compositions and gene expression profiles. These findings provide novel insights into the spatial heterogeneity and progression mechanisms of mucinous colorectal carcinoma, potentially informing future therapeutic strategies.
利益披露 Disclosure
H. Sada, None..
H. Niitsu, None..
H. Nakahara, None..
M. Miguchi, None..
H. Tashiro, None..
H. Ohdan, None..
T. Hinoi, None.