Kinomic profiling of glioblastoma cells reveals PLCG1 as a target in restricted glucose.
Kinomic profiling of glioblastoma cells reveals PLCG1 as a target in restricted glucose.
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DOI：
10.1186/s40364-018-0136-9
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发表时间：
2018
影响因子：
11.1
通讯作者：
Hjelmeland AB
中科院分区：
文献类型：
--
作者：
Walker K;Boyd NH;Anderson JC;Willey CD;Hjelmeland AB
为了使胶质母细胞瘤 (GBM) 治疗在体内有效，了解肿瘤微环境的影响势在必行。在传统的细胞培养条件下，葡萄糖浓度不能模拟生理水平，也不能模拟肿瘤微环境中浓度降低的情况。因此，我们试图分析限制性葡萄糖培养的 GBM 细胞中激酶活性的差异，以确定小分子抑制剂可以靶向的途径。使用 PamStation12 平台，我们检查了在标准或低葡萄糖条件下培养的细胞的 GBM 裂解物磷酸化与已知蛋白质磷酸化位点相对应的 144 个酪氨酸和 144 个丝氨酸/苏氨酸肽的能力。在 GBM 球体培养物中使用小分子激酶抑制剂鉴定和验证了潜在的激酶靶点。利用两个 GBM 患者来源的异种移植物的结果，我们确定了源自磷脂酶 C、Gamma 1 (PLCG1) 和 Raf-1 的肽的常见变化。使用PLC和Raf抑制剂，我们发现PLC抑制剂U73122在限制葡萄糖条件下具有明显更强的生长抑制作用。相比之下，无论测试的营养水平如何，Raf 抑制剂都具有显着的生长抑制作用。总之，我们的数据表明激酶活性在低葡萄糖条件下发生改变，并且激酶组学分析可以帮助确定针对 GBM 生长的有效策略。我们的数据进一步表明准确建模肿瘤微环境以重现癌细胞信号传导和开发抗癌药物筛选的重要性。
For glioblastoma (GBM) treatments to be effective in vivo, understanding the effects of the tumor microenvironment is imperative. In traditional cell culture conditions, glucose concentrations do not model physiologic levels, nor the diminished concentrations found in tumor niches. We therefore sought to profile the differences in kinase activity in GBM cells cultured in restricted glucose to identify pathways that could be targeted with small molecule inhibitors. Using the PamStation12 platform, we examined the ability of GBM lysates from cells cultured in standard or low glucose conditions to phosphorylate 144 tyrosine and 144 serine/threonine peptides that correspond to known protein phosphorylation sites. Potential kinase targets were identified and validated using small molecule kinase inhibitors in GBM spheroid cultures. Using results from two GBM patient-derived xenografts, we determined common changes to peptides derived from Phospholipase C, Gamma 1 (PLCG1) and Raf-1. Using PLC and Raf inhibitors, we found a significantly stronger growth inhibitory effect of the PLC inhibitor U73122 under restricted glucose conditions. In contrast, Raf inhibitors were significantly growth inhibitory regardless of the nutrient level tested. Together, our data demonstrate that kinase activity is altered in low glucose conditions and that kinomic profiling can assist with the identification of effective strategies to target GBM growth. Our data further suggest the importance of accurately modeling the tumor microenvironment to reproduce cancer cell signaling and develop drug screens for anti-cancer agents.
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DOI：
10.4161/cc.8.20.9701
发表时间：
2009-10-15
期刊：
Cell cycle (Georgetown, Tex.)
影响因子：
--
作者：
Heddleston JM;Li Z;McLendon RE;Hjelmeland AB;Rich JN
通讯作者：
Rich JN
DOI：
10.1158/1541-7786.mcr-14-0106-t
发表时间：
2014-11
期刊：
Molecular cancer research : MCR
影响因子：
--
作者：
Abbadi S;Rodarte JJ;Abutaleb A;Lavell E;Smith CL;Ruff W;Schiller J;Olivi A;Levchenko A;Guerrero-Cazares H;Quinones-Hinojosa A
通讯作者：
Quinones-Hinojosa A
影响因子：
158.5
作者：
Stupp, R;Mason, WP;Ryan, G
通讯作者：
Ryan, G
影响因子：
15.9
作者：
Vartanian, Alenoush;Singh, Sanjay K.;Zadeh, Gelareh
通讯作者：
Zadeh, Gelareh
影响因子：
5.3
作者：
Ferretti M;Fabbiano C;Di Bari M;Conte C;Castigli E;Sciaccaluga M;Ponti D;Ruggieri P;Raco A;Ricordy R;Calogero A;Tata AM
通讯作者：
Tata AM