PO.IM02.01 · 免疫学
构建重现转移性子宫内膜癌的免疫功能健全原位小鼠模型
Development of an immunocompetent orthotopic mouse model recapitulating metastatic endometrial cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
子宫内膜癌（EC）是最常见的妇科恶性肿瘤。然而，由于缺乏能够忠实重现人类EC（包括完整免疫应答和形成远处转移能力）的免疫功能健全动物模型，新型治疗策略的开发进展受限。为填补这一空白，我们使用GFP标记的小鼠子宫内膜癌细胞系MECPK（Pten缺失、Kras激活）构建了一个原位、免疫功能健全的EC小鼠模型。在子宫内膜刮伤后，将50,000个MECPK细胞注入受体小鼠的子宫腔内。使用荧光引导解剖显微镜，在诱导后第4、5、6和8周收集患EC的子宫组织。样本进行苏木精-伊红（H&E）染色和E-cadherin免疫组化处理，随后进行基于AI的图像分析。肿瘤起始在诱导后第5周首次可检测到，66.7%的小鼠（6/9）在第8周发生远处肺转移。所有荷瘤小鼠均在移植后10周内死于疾病。我们对PD-L1、MLH1和MSH2进行了定量IHC分析，以评估与免疫治疗应答相关的生物标志物。PD-L1在肿瘤进展的所有阶段均高表达。虽然MLH1表达在EC与正常区域之间无显著差异，但MSH2表达在癌症进展过程中显著降低。第5周原发癌中的MSH2表达（246.55±6.39）与正常上皮（256.15±10.96）无显著差异。然而，MSH2水平在第6周原发癌（95.65±17.88，p < 0.001）和第8周原发癌（133.16±12.98，p < 0.001）以及第8周肺内转移性子宫内膜癌（110.89±19.26，p < 0.001）中均显著降低。这些发现确立了一个稳健的免疫功能健全原位EC模型，重现了原发肿瘤进展和远处转移，为机制研究和包括免疫治疗在内的新兴疗法的临床前评估提供了宝贵平台。
本研究得到NCI R01 CA264944的资助。
查看英文原文 English abstract
Endometrial cancer (EC) is the most common gynecologic malignancy. However, progress in developing new therapeutic strategies has been limited by the lack of immunocompetent animal models that faithfully recapitulate human EC, including intact immune responses and the ability to form distant metastases. To address this gap, we developed an orthotopic, immunocompetent EC mouse model using the GFP-labeled mouse endometrial cancer cell line MECPK (Pten-deleted, Kras-activated). Following uterine abrasion, 50,000 MECPK cells were injected into the uterine lumen of recipient mice. Uterus tissues with EC were collected from mice at 4, 5, 6, and 8 weeks post-induction using a fluorescence-guided dissecting microscope. Samples were processed for hematoxylin and eosin (H&E) staining and immunohistochemistry for E-cadherin, followed by AI-based image analysis. Tumor initiation was first detectable at 5 weeks after induction, and 66.7% of mice (6/9) developed distant lung metastases by 8 weeks. All tumor-bearing mice succumbed to disease before 10 weeks post-transplantation. We performed quantitative IHC analysis of PD-L1, MLH1, and MSH2 to evaluate biomarkers associated with immunotherapy response. PD-L1 was highly expressed at all stages of tumor progression. While MLH1 expression did not differ significantly between EC and normal regions, MSH2 expression was remarkably reduced during cancer progression. MSH2 expression in primary cancer at 5 weeks (246.55 ±6.39) was not significantly different from that in normal epithelium (256.15 ±10.96). However, MSH2 levels were significantly decreased in primary cancer at 6 weeks (95.65 ±17.88, p <0.001) and 8 weeks (133.16 ±12.98, p <0.001), as well as in metastasis endometrial cancer in lung at 8 weeks (110.89 ±19.26, p <0.001). These findings establish a robust immunocompetent orthotopic EC model that recapitulates primary tumor progression and distant metastasis, providing a valuable platform for mechanistic studies and preclinical evaluation of emerging therapies, including immunotherapy.
This work was supported by NCI R01 CA264944
利益披露 Disclosure
K. Kim, None..
C. Schmidt, None..
E. Jeong, None..
T. Kim, None..
J. Jeong, None.