PO.TB03.04 · 肿瘤生物学
光学成像与数字病理学评估提升患者来源转移性乳腺癌模型的转化价值与稳健性
Optical imaging and digital pathology evaluation increases the translational value and robustness of patient-derived metastatic breast cancer models
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摘要 Abstract
中文摘要
乳腺癌仍然是女性最常见的恶性肿瘤，也是癌症相关死亡的主要原因，这主要归因于转移性进展。能够重现转移级联过程的可靠临床前模型对于理解疾病机制和评估新型疗法至关重要。传统方法虽具信息价值，但往往缺乏在体内追踪转移扩散所需的敏感性和动态分辨率。本研究展示了将体内光学数字图像分析与组织病理学数字图像分析相结合，以增强患者来源异种移植（PDX）转移性乳腺癌模型的检测、表征和转化相关性的效用。首先，使用两种实验方法在体外对三种乳腺癌模型的转移潜能进行分类：2D划痕愈合实验和3D球体侵袭实验。随后，建立了五个PDX模型，包括两个Her2+模型（1162、1322）和三个三阴性模型（401、857、1387），并评估其在小鼠体内的自发转移。乳腺癌细胞系MDAMB231作为阳性对照。为实现体内追踪，肿瘤细胞被感染荧光报告基因或生物发光报告基因（荧光素酶），并使用光学成像系统（IVIS Lumina S5和Licor Pearl）进行可视化。在体外实验中，MDAMB231、MCF-7和401（三阴性PDX）分别被分类为高转移、低转移和无转移。在体内肺转移实验中，两个三阴性模型（401、1387）和一个HER2+模型（1387）被分类为低转移。另一个三阴性模型（1162）和HER2+模型（1162）则完全未显示转移。遗憾的是，由于报告基因随传代次数增加而逐渐减弱，无法建立稳定表达报告基因的PDX。然而，这些结果表明瞬时转染是在体内追踪肿瘤生长的一种有前景的工具。最后，收获器官，并通过组织学（H&E）和人特异性免疫组化（IHC，hLaminB1）评估转移灶。hLaminB1+转移灶使用Visiopharm软件通过数字图像分析（DIA）进行定量。MDAMB231主要转移至肺，较小程度转移至肝和脑。其转移水平远高于所有PDX，反映了该细胞系的侵袭性生长。DIA为PDX模型提供了稳健的解读，表明这一整合工作流程有助于转移模型的选择和定量，在治疗测试和机制研究中具有应用价值。
查看英文原文 English abstract
Breast cancer remains the most prevalent malignancy among women and a leading cause of cancer-related death, primarily due to metastatic progression. Reliable preclinical models that recapitulate the metastatic cascade are essential for understanding disease mechanisms and evaluating novel therapies. Traditional methods, while informative, often lack the sensitivity and dynamic resolution needed to track metastatic spread in vivo . This study demonstrates the utility of integrating in vivo optical digital image analysis with histopathological digital image analysis to enhance the detection, characterization, and translational relevance of patient-derived xenograft (PDX) models of metastatic breast cancer. The metastatic potential of three breast cancer models first was classified in-vitro using two assays: 2D scratch wound and 3D spheroid invasion. Next, five PDX models, composed of two Her2+(1162, 1322) and three triple negative models (401, 857, 1387), were developed and evaluated for spontaneous in-vivo metastasis in mice. The breast cancer cell line MDAMB231 served as positive control. To enable in vivo tracking, tumor cells were infected with a fluorescent reporter or a bioluminescent reporter (luciferase) and visualized using optical imaging systems (IVIS Lumina S5 and Licor Pearl). In the in-vitro assays, MDAMB231, MCF-7, and 401 (triple negative PDX) were classified as high, low, and non-metastatic respectively. In the in-vivo lung metastasis assay, two triple negative models (401, 1387) and one HER2+ model (1387) were classified as low-metastatic. The other triple negative model (1162) and HER2+ model (1162) displayed no metastases at all. Unfortunately, it was not possible to create a PDX with a reporter that was stably expressed because the reporters reduced with passages over time. However, these results demonstrate that transient transfection is a promising tool to follow tumor growth in vivo. Finally, the organs were harvested, and metastases were evaluated via histology (H&E) and human-specific immunohistochemistry (IHC, hLaminB1). The hLaminB1+ metastasis were quantified by digital image analysis (DIA) using the Visiopharm software. MDAMB231 metastasized mainly to the lung and to a lower extend to the liver and the brain. Its metastatic level was much higher than all the PDXs, reflecting the aggressive growth of the cell line. The DIA provided a robust interpretation of the PDX models, illustrating that this integrated workflow supports the selection and quantification of metastatic models, with applications in therapeutic testing and mechanistic studies.
利益披露 Disclosure
P. Meyer, None..
L. Neagu-Lund, None..
E. Oswald, None..
A. Zuraw, None..
L. Weichert, None..
M. Staup, None..
J. B. Schueler, None.