Strekalova Elena, Malin Dmitry, Rajanala Harisha, Cryns Vincent L
Department of Medicine, University of Wisconsin Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
Methods Mol Biol. 2019;1866:61-73. doi: 10.1007/978-1-4939-8796-2_6.
We have developed a novel therapeutic paradigm ("metabolic priming") for cancer whereby restriction of the essential amino acid methionine activates a number of cell-stress-response pathways that can be selectively targeted to enhance the therapeutic impact of methionine restriction. One example of metabolic priming is the combination of methionine restriction with proapoptotic TRAIL receptor-2 (TRAIL-R2) agonists. Methionine restriction enhances the cell surface expression of TRAIL-R2 selectively in transformed breast epithelial cells and renders them more susceptible to cell death induction by TRAIL-R2 agonists in cellular and murine models of breast cancer. This methods review focuses on preclinical models of breast cancer to investigate metabolic priming by methionine restriction. Multiple cell-based methods are detailed to measure cell viability, cell survival, caspase activity, apoptosis, and matrix detachment-induced cell death (anoikis). In addition, we describe an orthotopic model of metastatic breast cancer that utilizes mCherry-fluorescently-labeled human breast cancer cells. This model captures the entire metastatic cascade from the mammary gland to the lung and mimics key features of the human disease. These breast-cancer models can be readily adapted to other tumor types. Overall, we provide a stepwise, translationally-relevant approach to study metabolic priming in the context of cancer.
我们已经开发出一种针对癌症的新型治疗模式（“代谢预激”），即限制必需氨基酸甲硫氨酸可激活多种细胞应激反应途径，这些途径可被选择性靶向，以增强甲硫氨酸限制的治疗效果。代谢预激的一个例子是甲硫氨酸限制与促凋亡的肿瘤坏死因子相关凋亡诱导配体受体-2（TRAIL-R2）激动剂联合使用。在乳腺癌的细胞和小鼠模型中，甲硫氨酸限制可选择性地增强转化乳腺上皮细胞表面TRAIL-R2的表达，并使它们对TRAIL-R2激动剂诱导的细胞死亡更敏感。本方法综述聚焦于乳腺癌的临床前模型，以研究通过甲硫氨酸限制进行的代谢预激。详细介绍了多种基于细胞的方法，以测量细胞活力、细胞存活、半胱天冬酶活性、凋亡以及基质脱离诱导的细胞死亡（失巢凋亡）。此外，我们描述了一种转移性乳腺癌的原位模型，该模型利用了mCherry荧光标记的人乳腺癌细胞。该模型捕捉了从乳腺到肺的整个转移级联过程，并模拟了人类疾病的关键特征。这些乳腺癌模型可很容易地应用于其他肿瘤类型。总体而言，我们提供了一种逐步的、与转化相关的方法，用于在癌症背景下研究代谢预激。
