Targeted Mass Spectrometry Enables Quantification of Novel Pharmacodynamic Biomarkers of ATM Kinase Inhibition.
Targeted Mass Spectrometry Enables Quantification of Novel Pharmacodynamic Biomarkers of ATM Kinase Inhibition.
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靶向质谱法能够定量ATM激酶抑制的新型药效学生物标志物。
DOI：
10.3390/cancers13153843
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发表时间：
2021-07-30
期刊：
影响因子：
5.2
通讯作者：
Paulovich AG
中科院分区：
文献类型：
--
作者：
Whiteaker JR;Wang T;Zhao L;Schoenherr RM;Kennedy JJ;Voytovich U;Ivey RG;Huang D;Lin C;Colantonio S;Caceres TW;Roberts RR;Knotts JG;Kaczmarczyk JA;Blonder J;Reading JJ;Richardson CW;Hewitt SM;Garcia-Buntley SS;Bocik W;Hiltke T;Rodriguez H;Harrington EA;Barrett JC;Lombardi B;Marco-Casanova P;Pierce AJ;Paulovich AG
细胞DNA损伤反应（DDR）网络的功能影响癌症发生的风险，DDR也是癌症治疗的靶点。因此，我们有可靠的实验室方法来确定这个网络的活动是很重要的。我们描述了基于靶向质谱的51-plex测定（DDR-2）的开发和分析验证，用于测量与DDR相关的蛋白质和翻译后修饰。这些发现证明了潜在的新型药效学生物标志物的鉴定。ATM丝氨酸/苏氨酸激酶（HGNC: ATM）参与DNA双链断裂修复的启动，ATM抑制剂目前正在临床试验中作为抗癌药物进行测试，其中药效学（PD）分析对于帮助指导剂量和计划以及支持作用机制研究至关重要。为了鉴定和量化ATM抑制的PD生物标志物，我们开发并分析验证了51-plex测定法（DDR-2），定量蛋白质表达和DNA损伤反应磷酸化。定量下限的中位数为1.28 fmol，线性范围在3个数量级以上，测定间变异的中位数为11% CV， 86%的多肽在分析前可以稳定保存。实验证明，使用该方法可以量化电离辐射诱导的永生化淋巴母细胞系和原代人外周血单个核细胞DNA损伤后的信号传导，鉴定出抑制ATM的PD生物标志物，以支持临床前和临床研究。
Functionality of the cellular DNA damage response (DDR) network affects risk for developing cancer, and the DDR is also a target of cancer therapies. Thus, it is important that we have reliable laboratory methods for determining the activity of this network. We describe the development and analytical validation of a targeted mass spectrometry-based 51-plex assay (DDR-2) for measuring proteins and post-translational modifications related to the DDR. The findings demonstrate identification of potential novel pharmacodynamic biomarkers. The ATM serine/threonine kinase (HGNC: ATM) is involved in initiation of repair of DNA double-stranded breaks, and ATM inhibitors are currently being tested as anti-cancer agents in clinical trials, where pharmacodynamic (PD) assays are crucial to help guide dose and scheduling and support mechanism of action studies. To identify and quantify PD biomarkers of ATM inhibition, we developed and analytically validated a 51-plex assay (DDR-2) quantifying protein expression and DNA damage-responsive phosphorylation. The median lower limit of quantification was 1.28 fmol, the linear range was over 3 orders of magnitude, the median inter-assay variability was 11% CV, and 86% of peptides were stable for storage prior to analysis. Use of the assay was demonstrated to quantify signaling following ionizing radiation-induced DNA damage in both immortalized lymphoblast cell lines and primary human peripheral blood mononuclear cells, identifying PD biomarkers for ATM inhibition to support preclinical and clinical studies.
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影响因子：
120.1
作者：
Cook, David;Brown, Dearg;Pangalos, Menelas N.
通讯作者：
Pangalos, Menelas N.
影响因子：
16
作者：
Liu S;Shiotani B;Lahiri M;Maréchal A;Tse A;Leung CC;Glover JN;Yang XH;Zou L
通讯作者：
Zou L
影响因子：
7
作者：
Kennedy, Jacob J.;Yan, Ping;Paulovich, Amanda G.
通讯作者：
Paulovich, Amanda G.
影响因子：
9.3
作者：
Kushnir MM;Rockwood AL;Roberts WL;Abraham D;Hoofnagle AN;Meikle AW
通讯作者：
Meikle AW
影响因子：
9.9
作者：
Lange, Vinzenz;Picotti, Paola;Domon, Bruno;Aebersold, Ruedi
通讯作者：
Aebersold, Ruedi