Colony Lysate Arrays for Proteomic Profiling of Drug-Tolerant Persisters of Cancer Cell.
Colony Lysate Arrays for Proteomic Profiling of Drug-Tolerant Persisters of Cancer Cell.
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DOI：
10.1021/acs.analchem.7b01215
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发表时间：
2017-08
影响因子：
7.4
通讯作者：
K. Kume;S. Nishizuka
中科院分区：
文献类型：
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作者：
K. Kume;S. Nishizuka
癌细胞的功能异质性是了解药物治疗后复发的关键属性之一。因此，需要澄清哪些类型的癌细胞亚群主要导致复发。最近，我们建立了菌落裂解物阵列(Cola)，这是一种基于反相蛋白质阵列(RPPA)系统的方法，允许在蛋白质水平上比较单个菌落来评估抗癌药物耐药持续体(DTP)的启动。DTP在不同的药物浓度和类型下生长，在平面上呈二维生长(∼为1 mm)。在琼脂糖凝胶中，DTP的大小比球形(∼0.3 mm)大，这使得它们易于处理，可用于多种分析。DTP在其形成过程中提供功能信息，从耐药单细胞的起源开始。使用从不同药物产生的DTP和基于44种蛋白质的剂量分布，我们证明了DTP是基于它们的蛋白质组学随着药物和剂量的变化而变化而聚集在一起的。有趣的是，DTP中的9个转录因子，如STAT3和Oct4A，被鉴定为对吉非替尼的反应降低或增加了蛋白质水平。重要的是，这些结果只能通过个体蛋白质组菌落图谱获得，这可能确定DTP的替代治疗靶点和生物标记物，这些靶点和生物标记物可能包含癌症复发的关键机制。
Functional heterogeneity of cancer cells is one of the key properties to understanding relapse after drug treatment. Hence, clarification is needed with regard to which types of subgroups of cancer cells dominantly contribute to the initiation of relapse. Recently, we established the colony lysate array (CoLA), which is a method that allows comparison of individual colonies at the protein level to assess the initiation of anticancer drug-tolerant persisters (DTPs) based on the reverse-phase protein array (RPPA) system. DTPs grow in various drug concentrations and types showing 2-dimensional growth (∼1 mm) on a flat surface. The size of DTPs are larger than spheroids (∼0.3 mm) in agarose gel, which makes them easy to handle for a number of assays. DTPs provide functional information during the process of their formation, initiating from the origin of a drug-tolerant single cell. Using >2000 DTPs generated from various drugs and doses profiled on the basis of 44 proteins, we demonstrate that the DTPs are clustered on the basis of their proteomic profiles changing in response to drugs and doses. Of interest, nine transcription factors in the DTPs, such as STAT3 and OCT4A, were identified as having decreased or increased levels of proteins in response to gefitinib. Importantly, these results can be obtained only by individual proteomic colony profiling, which may identify alternative therapeutic targets and biomarkers for DTPs that may harbor critical mechanisms for cancer relapse.