Bilateral murine tumor models for characterizing the response to immune checkpoint blockade.,Nature Protocols

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Bilateral murine tumor models for characterizing the response to immune checkpoint blockade.
Nature Protocols ( IF 13.1 ) Pub Date : 2020-04-01 , DOI: 10.1038/s41596-020-0299-3
Rachael M Zemek _{1,

2,

3} , Vanessa S Fear _{1,

2,

3} , Cath Forbes _{1,

2,

3} , Emma de Jong ₃ , Thomas H Casey ₂ , Louis Boon ₄ , Timo Lassmann ₃ , Anthony Bosco ₃ , Michael J Millward _{2,

5,

6} , Anna K Nowak _{2,

5,

6} , Richard A Lake _{1,

2} , W Joost Lesterhuis _{1,

2,

3}

Affiliation

The therapeutic response to immune checkpoint blockade (ICB) is highly variable, not only between different cancers but also between patients with the same cancer type. The biological mechanisms underlying these differences in response are incompletely understood. Identifying correlates in patient tumor samples is challenging because of genetic and environmental variability. Murine studies usually compare different tumor models or treatments, introducing potential confounding variables. This protocol describes bilateral murine tumor models, derived from syngeneic cancer cell lines, that display a symmetrical yet dichotomous response to ICB. These models enable detailed analysis of whole tumors in a highly homogeneous background, combined with knowledge of the therapeutic outcome within a few weeks, and could potentially be used for mechanistic studies using other (immuno-)therapies. We discuss key considerations and describe how to use two cell lines as fully optimized models. We discuss experimental details, including proper inoculation technique to achieve symmetry and one-sided surgical tumor removal, which takes only 5 min per mouse. Furthermore, we outline the preparation of bulk tissue or single-cell suspensions for downstream analyses such as bulk RNA-seq, immunohistochemistry, single-cell RNA-seq and flow cytometry.

中文翻译：

用于表征对免疫检查点阻断反应的双侧鼠肿瘤模型。

对免疫检查点阻断 (ICB) 的治疗反应是高度可变的，不仅在不同癌症之间，而且在具有相同癌症类型的患者之间。这些反应差异背后的生物学机制尚不完全清楚。由于遗传和环境变异性，识别患者肿瘤样本中的相关性具有挑战性。鼠类研究通常比较不同的肿瘤模型或治疗，引入潜在的混杂变量。该协议描述了源自同基因癌细胞系的双侧鼠肿瘤模型，该模型对 ICB 表现出对称但二分法的反应。这些模型能够在高度同质的背景下详细分析整个肿瘤，并在几周内结合对治疗结果的了解，并且可能用于使用其他（免疫）疗法的机制研究。我们讨论了关键考虑因素并描述了如何使用两个细胞系作为完全优化的模型。我们讨论了实验细节，包括适当的接种技术以实现对称性和单侧手术肿瘤切除，每只小鼠仅需 5 分钟。此外，我们概述了用于下游分析的大块组织或单细胞悬液的制备，例如大块 RNA-seq、免疫组织化学、单细胞 RNA-seq 和流式细胞术。

更新日期：2020-04-01

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