An integrated method for single cell proteomics with simultaneous measurements of intracellular drug concentration implicates new mechanisms for adaptation to KRASG12D inhibitors.
An integrated method for single cell proteomics with simultaneous measurements of intracellular drug concentration implicates new mechanisms for adaptation to KRASG12D inhibitors.
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一种同时测量细胞内药物浓度的单细胞蛋白质组学集成方法暗示了适应 KRASG12D 抑制剂的新机制。
DOI：
10.1101/2023.11.18.567669
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Orsburn,BenjaminC
中科院分区：
文献类型：
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作者：
Orsburn,BenjaminC
已经确定的是，单个人类细胞群体通常会以异质性方式对相同的药物治疗作出反应。在化疗的背景下，这些不同的反应可能会导致个体的适应机制，并最终导致多种不同的耐药方法。从药理学的角度来看，一个明显的问题是活性药物的细胞内浓度在单个细胞之间是如何变化的，这种变化在药物反应异质性中起着什么作用？迄今为止，还没有描述用于快速测量细胞内药物水平同时测量药物反应的集成方法。本研究描述了一种单细胞制备方法，该方法允许从单细胞中提取和消化蛋白质，同时保持同时测量小分子的条件。所述方法允许每天每个仪器分析多达40个细胞。当应用于KRASG12D小分子抑制剂时，我观察到广泛程度的药物细胞内水平，并且蛋白质组学反应主要基于每个单细胞内的药物浓度分层。进一步的工作正在进行中，以开发和标准化这种方法，更重要的是，标准化药物测量对细胞体积的直接测量。然而，这些初步结果似乎有希望识别具有独特药物反应机制的单细胞。本研究中描述的所有数据已通过ProteomeXchange联盟以登录号PXD 046002公开提供。
It is well established that a population of single human cells will often respond to the same drug treatment in a heterogeneous manner. In the context of chemotherapeutics, these diverse responses may lead to individual adaptation mechanisms and ultimately multiple distinct methods of resistance. The obvious question from a pharmacology perspective is how intracellular concentrations of active drug varies between individual cells, and what role does that variation play in drug response heterogeneity? To date, no integrated methods for rapidly measuring intracellular drug levels while simultaneously measuring drug responses have been described. This study describes a method for single cell preparation that allows proteins to be extracted and digested from single cells while maintaining conditions for small molecules to be simultaneously measured. The method as described allows up to 40 cells to be analyzed per instrument per day. When applied to a KRASG12D small molecule inhibitor I observe a wide degree of intracellular levels of the drug, and that proteomic responses largely stratify based on the concentration of drug within each single cell. Further work is in progress to develop and standardize this method and – more importantly – to normalize drug measurements against direct measurements of cell volume. However, these preliminary results appear promising for the identification of single cells with unique drug response mechanisms. All data described in this study has been made publicly available through the ProteomeXchange consortium under accession PXD046002.