Activity-Based Protein Profiling Shows Heterogeneous Signaling Adaptations to BRAF Inhibition.
Activity-Based Protein Profiling Shows Heterogeneous Signaling Adaptations to BRAF Inhibition.
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DOI：
10.1021/acs.jproteome.6b00613
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发表时间：
2016-12-02
影响因子：
4.4
通讯作者：
Koomen JM
中科院分区：
文献类型：
--
作者：
Sharma R;Fedorenko I;Spence PT;Sondak VK;Smalley KS;Koomen JM
BRAF V600 E突变型黑色素瘤患者通常用靶向BRAF激酶抑制剂治疗，如维罗非尼和达拉非尼。虽然这些药物最初是有效的，但它们不是治愈性的。迄今为止，大多数的焦点都集中在获得性抗性的遗传机制上;因此，我们必须更好地了解介导BRAF抑制逃逸的全球信号适应。在目前的研究中，我们使用基于活性的蛋白质谱（ABPP）与ATP类似物探针，以丰富激酶和其他酶类，有助于BRAF抑制剂（BRAFi）耐药在四个配对的同基因BRAFi初治/耐药细胞系模型。我们的分析表明，这些细胞系模型，这在他们的PTEN状态不同，有相当大的异质性，在他们的激酶ATP探针摄取比较幼稚细胞和适应慢性药物暴露。包括FAK 1、SLK和TAOK 2在内的许多激酶在BRAFi抗性细胞中增加了ATP探针摄取，而KHS 1（M4 K5）和BRAF在BRAFi抗性细胞中降低了ATP探针摄取。基因本体（GO）富集分析显示，BRAFi抗性与涉及细胞骨架组织和粘附的蛋白质中ATP探针摄取的显著增强以及与细胞代谢过程相关的蛋白质中ATP探针摄取的降低相关。ABPP方法能够鉴定对每个BRAFi抗性细胞系至关重要的关键表型介质。总之，这些数据表明，BRAF抑制的常见表型适应可以通过非常不同的信号传导网络介导，这表明BRAF突变黑素瘤细胞的信号传导中存在相当大的冗余。
Patients with BRAF V600E mutant melanoma are typically treated with targeted BRAF kinase inhibitors, such as vemurafenib and dabrafenib. Although these drugs are initially effective, they are not curative. Most of the focus to date has been upon genetic mechanisms of acquired resistance; therefore, we must better understand the global signaling adaptations that mediate escape from BRAF inhibition. In the current study, we have used activity-based protein profiling (ABPP) with ATP-analogue probes to enrich kinases and other enzyme classes that contribute to BRAF inhibitor (BRAFi) resistance in four paired isogenic BRAFi-naïve/resistant cell line models. Our analysis showed these cell line models, which differ in their PTEN status, have considerable heterogeneity in their kinase ATP probe uptake in comparing naïve cells and adaptations to chronic drug exposure. A number of kinases including FAK1, SLK and TAOK2 had increased ATP probe uptake in BRAFi resistant cells, while KHS1 (M4K5) and BRAF had decreased ATP probe uptake in the BRAFi-resistant cells. Gene ontology (GO) enrichment analysis revealed BRAFi resistance is associated with a significant enhancement in ATP probe uptake in proteins implicated in cytoskeletal organization and adhesion, and decreases in ATP probe uptake in proteins associated with cell metabolic processes. The ABPP approach was able to identify key phenotypic mediators critical for each BRAFi resistant cell line. Together, these data show that common phenotypic adaptations to BRAF inhibition can be mediated through very different signaling networks, suggesting considerable redundancy within the signaling of BRAF mutant melanoma cells.
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影响因子：
8
作者：
Fedorenko IV;Abel EV;Koomen JM;Fang B;Wood ER;Chen YA;Fisher KJ;Iyengar S;Dahlman KB;Wargo JA;Flaherty KT;Sosman JA;Sondak VK;Messina JL;Gibney GT;Smalley KS
通讯作者：
Smalley KS
影响因子：
4.4
作者：
Cox, Juergen;Neuhauser, Nadin;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
16.6
作者：
Long, Georgina V.;Fung, Carina;Rizos, Helen
通讯作者：
Rizos, Helen
影响因子：
2.7
作者：
Chartier M;Chénard T;Barker J;Najmanovich R
通讯作者：
Najmanovich R
影响因子：
64.8
作者：
Johannessen, Cory M.;Boehm, Jesse S.;Kim, So Young;Thomas, Sapana R.;Wardwell, Leslie;Johnson, Laura A.;Emery, Caroline M.;Stransky, Nicolas;Cogdill, Alexandria P.;Barretina, Jordi;Caponigro, Giordano;Hieronymus, Haley;Murray, Ryan R.;Salehi-Ashtiani, Kourosh;Hill, David E.;Vidal, Marc;Zhao, Jean J.;Yang, Xiaoping;Alkan, Ozan;Kim, Sungjoon;Harris, Jennifer L.;Wilson, Christopher J.;Myer, Vic E.;Finan, Peter M.;Root, David E.;Roberts, Thomas M.;Golub, Todd;Flaherty, Keith T.;Dummer, Reinhard;Weber, Barbara L.;Sellers, William R.;Schlegel, Robert;Wargo, Jennifer A.;Hahn, William C.;Garraway, Levi A.
通讯作者：
Garraway, Levi A.