Profiling phospho-signaling networks in breast cancer using reverse-phase protein arrays.
Profiling phospho-signaling networks in breast cancer using reverse-phase protein arrays.
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DOI：
10.1038/onc.2012.378
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发表时间：
2013-07-18
期刊：
影响因子：
8
通讯作者：
Sorger, P.
中科院分区：
文献类型：
--
作者：
Gujral, T. S.;Karp, R. L.;Finski, A.;Chan, M.;Schwartz, P. E.;MacBeath, G.;Sorger, P.
测量肿瘤样本中细胞信号传导通路的状态有望促进对肿瘤发生的理解并识别反应生物标志物。在这里，我们描述了使用反相蛋白质阵列（RPPA或RPLA）来分析56例乳腺癌和匹配的正常组织中的信号蛋白。在RPPA中，数百到数千个裂解物排列在密集的规则网格中，每个网格用不同的抗体探测（目前的工作中有100个，其中71个产生了乳腺组织的强信号）。虽然RPPA技术被广泛使用，但测量反映蛋白质活化的磷酸化变化仍然具有挑战性。使用重复沉积和经过充分验证的抗体，我们表明可以在肿瘤样品中监测不同的磷酸化模式，并以连贯的方式将变化映射到信号网络上。这些模式与基于生物标志物的乳腺癌分类和已知的肿瘤发生机制一致。我们详细探讨了一种肿瘤相关的模式，涉及Axl受体酪氨酸激酶（RTK）和cMet RTK磷酸化丰度的变化。cMet和Axl都与乳腺癌或对抗癌药物的耐药性有关，但这两种RTK在功能上并不相关。在“三阴性”乳腺细胞系中的蛋白质消耗和过表达研究揭示了Axl和cMet之间的串扰，其涉及Axl介导的cMet修饰，这是Axl配体Gas 6在有效和及时的信号转导中对cMet的要求，以及两种受体物理相互作用的潜力。这些发现具有潜在的治疗意义，因为它们意味着双特异性受体抑制剂（例如ATP竞争性小激酶抑制剂，如GSK 1363089、BMS-777607或MP 470）可能比目前正在开发的单特异性治疗抗体（例如MetMAb）更有效。
Measuring the states of cell signaling pathways in tumor samples promises to advance understanding of oncogenesis and identify response biomarkers. Here, we describe the use of Reverse Phase Protein Arrays (RPPAs or RPLAs) to profile signaling proteins in 56 breast cancers and matched normal tissue. In RPPAs, hundreds to thousands of lysates are arrayed in dense regular grids and each grid is probed with a different antibody (100 in the current work, of which 71 yielded strong signals with breast tissue). Although RPPA technology is quite widely used, measuring changes in phosphorylation reflective of protein activation remains challenging. Using repeat deposition and well-validated antibodies we show that diverse patterns of phosphorylation can be monitored in tumor samples and changes mapped onto signaling networks in a coherent fashion. The patterns are consistent with biomarker-based classification of breast cancers and known mechanisms of oncogenesis. We explore in detail one tumor-associated pattern that involves changes in the abundance of the Axl receptor tyrosine kinase (RTK) and phosphorylation of the cMet RTK. Both cMet and Axl have been implicated in breast cancer, or in resistance to anti-cancer drugs, but the two RTKs are not known to be linked functionally. Protein depletion and over-expression studies in a “triple-negative” breast cells line reveal crosstalk between Axl and cMet involving Axl-mediated modification of cMet, a requirement for cMet in efficient and timely signal transduction by the Axl ligand Gas6 and the potential for the two receptors to interact physically. These findings have potential therapeutic implications since they imply that bi-specific receptor inhibitors (e.g. ATP-competitive small kinase inhibitors such as GSK1363089, BMS-777607 or MP470) may be more efficacious than the monospecific therapeutic antibodies currently in development (e.g. MetMAb).
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影响因子：
4.8
作者：
Demarchi, F;Verardo, R;Schneider, C
通讯作者：
Schneider, C
影响因子：
3.8
作者：
Gonzalez-Angulo AM;Hennessy BT;Meric-Bernstam F;Sahin A;Liu W;Ju Z;Carey MS;Myhre S;Speers C;Deng L;Broaddus R;Lluch A;Aparicio S;Brown P;Pusztai L;Symmans WF;Alsner J;Overgaard J;Borresen-Dale AL;Hortobagyi GN;Coombes KR;Mills GB
通讯作者：
Mills GB
影响因子：
8
作者：
Mahadevan, D.;Cooke, L.;Bearss, D.
通讯作者：
Bearss, D.
影响因子：
64.8
作者：
Marcotte, EM;Pellegrini, M;Eisenberg, D
通讯作者：
Eisenberg, D
DOI：
10.1073/pnas.0909333107
发表时间：
2010-01-19
影响因子：
11.1
作者：
Gjerdrum, Christine;Tiron, Crina;Lorens, James B.
通讯作者：
Lorens, James B.