Yap governs a lineage-specific neuregulin1 pathway-driven adaptive resistance to RAF kinase inhibitors.
Yap governs a lineage-specific neuregulin1 pathway-driven adaptive resistance to RAF kinase inhibitors.
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DOI：
10.1186/s12943-022-01676-9
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发表时间：
2022-12-07
期刊：
影响因子：
37.3
通讯作者：
Fagin, James A. A.
中科院分区：
文献类型：
--
作者：
Garcia-Rendueles, Maria E. R.;Krishnamoorthy, Gnana;Saqcena, Mahesh;Acuna-Ruiz, Adrian;Revilla, Giovanna;de Stanchina, Elisa;Knauf, Jeffrey A. A.;Lester, Rona;Xu, Bin;Ghossein, Ronald A. A.;Fagin, James A. A.
Hippo 通路的失活会促进 Yap 核易位，从而能够执行诱导组织生长的转录程序。 Hippo 中间体的基因损伤只能识别少数具有非法 YAP 激活的癌症。 Yap 与靶向治疗耐药有关，但 YAP 影响 MAPK 抑制剂适应性耐药的机制尚不清楚。我们通过免疫荧光和细胞裂解物的分级筛选了 52 个甲状腺癌细胞系，以确定 YAP 的非法核定位。我们设计了强力霉素 (dox) 诱导的甲状腺特异性小鼠模型，该模型可单独表达或与 HrasG12V 或 BrafV600E 的内源表达组合表达组成型核 YAPS127A。我们还生成了表达 dox 诱导的 sh-miR-E-YAP 和/或 YAPS127A 的细胞系。我们使用细胞活力、侵袭测定、免疫荧光、蛋白质印迹、qRT-PCR、流式细胞术和细胞分选、高通量批量 RNA 测序和体内肿瘤发生来研究 BRAF 突变细胞对 YAP 依赖性和对维莫非尼的反应。我们发现，在高密度培养时，27/52 甲状腺癌细胞系 (NU-YAP) 具有组成性异常的 YAP 核定位，这使得它们的生存能力、侵袭性和对 YAP-TEAD 复合物抑制剂维替泊芬的敏感性依赖于 YAP，而汇合驱动的 YAP 核排斥细胞 (CYT-YAP) 则不然。用 RAF 激酶抑制剂处理 BRAF 突变型甲状腺癌细胞，导致 YAP 核易位并激活其转录输出。 BRAF 突变型甲状腺细胞对维莫非尼的耐药性是由所有测试的同基因人类和小鼠甲状腺细胞系中 YAP 依赖性 NRG1、HER2 和 HER3 激活驱动的，这种激活可通过沉默 YAP 来消除，并通过泛 HER 激酶抑制剂来缓解。 YAP 激活诱导了 BRAF 黑色素瘤的类似变化，但不诱导结直肠细胞。甲状腺癌中 YAP 的激活产生对该转录因子的依赖性。 YAP 控制对 RAF 激酶抑制剂的适应性抗性，并在 BRAFV600E 突变细胞中诱导基因表达程序，该程序包含 NRG1 信号通路中的效应子，该程序在以谱系依赖性方式对 MAPK 抑制剂不敏感中发挥核心作用。 HIPPO 通路失活充当谱系依赖性变阻器，控制 BRAF-V600E 癌症中 MAPK 抑制剂反馈反应的适应性缓解程度。在线版本包含可在 10.1186/s12943-022-01676-9 获取的补充材料。
Inactivation of the Hippo pathway promotes Yap nuclear translocation, enabling execution of a transcriptional program that induces tissue growth. Genetic lesions of Hippo intermediates only identify a minority of cancers with illegitimate YAP activation. Yap has been implicated in resistance to targeted therapies, but the mechanisms by which YAP may impact adaptive resistance to MAPK inhibitors are unknown. We screened 52 thyroid cancer cell lines for illegitimate nuclear YAP localization by immunofluorescence and fractionation of cell lysates. We engineered a doxycycline (dox)-inducible thyroid-specific mouse model expressing constitutively nuclear YAPS127A, alone or in combination with endogenous expression of either HrasG12V or BrafV600E. We also generated cell lines expressing dox-inducible sh-miR-E-YAP and/or YAPS127A. We used cell viability, invasion assays, immunofluorescence, Western blotting, qRT-PCRs, flow cytometry and cell sorting, high-throughput bulk RNA sequencing and in vivo tumorigenesis to investigate YAP dependency and response of BRAF-mutant cells to vemurafenib. We found that 27/52 thyroid cancer cell lines had constitutively aberrant YAP nuclear localization when cultured at high density (NU-YAP), which rendered them dependent on YAP for viability, invasiveness and sensitivity to the YAP-TEAD complex inhibitor verteporfin, whereas cells with confluency-driven nuclear exclusion of YAP (CYT-YAP) were not. Treatment of BRAF-mutant thyroid cancer cells with RAF kinase inhibitors resulted in YAP nuclear translocation and activation of its transcriptional output. Resistance to vemurafenib in BRAF-mutant thyroid cells was driven by YAP-dependent NRG1, HER2 and HER3 activation across all isogenic human and mouse thyroid cell lines tested, which was abrogated by silencing YAP and relieved by pan-HER kinase inhibitors. YAP activation induced analogous changes in BRAF melanoma, but not colorectal cells. YAP activation in thyroid cancer generates a dependency on this transcription factor. YAP governs adaptive resistance to RAF kinase inhibitors and induces a gene expression program in BRAFV600E-mutant cells encompassing effectors in the NRG1 signaling pathway, which play a central role in the insensitivity to MAPK inhibitors in a lineage-dependent manner. HIPPO pathway inactivation serves as a lineage-dependent rheostat controlling the magnitude of the adaptive relief of feedback responses to MAPK inhibitors in BRAF-V600E cancers. The online version contains supplementary material available at 10.1186/s12943-022-01676-9.
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河马效应子YAP促进了对RAF和MEK靶向的癌症疗法的耐药性。
影响因子：
30.8
作者：
Lin L;Sabnis AJ;Chan E;Olivas V;Cade L;Pazarentzos E;Asthana S;Neel D;Yan JJ;Lu X;Pham L;Wang MM;Karachaliou N;Cao MG;Manzano JL;Ramirez JL;Torres JM;Buttitta F;Rudin CM;Collisson EA;Algazi A;Robinson E;Osman I;Muñoz-Couselo E;Cortes J;Frederick DT;Cooper ZA;McMahon M;Marchetti A;Rosell R;Flaherty KT;Wargo JA;Bivona TG
通讯作者：
Bivona TG
影响因子：
64.5
作者：
Cancer Genome Atlas Research Network
通讯作者：
Cancer Genome Atlas Research Network
影响因子：
11.2
作者：
Ishii N;Harada N;Joseph EW;Ohara K;Miura T;Sakamoto H;Matsuda Y;Tomii Y;Tachibana-Kondo Y;Iikura H;Aoki T;Shimma N;Arisawa M;Sowa Y;Poulikakos PI;Rosen N;Aoki Y;Sakai T
通讯作者：
Sakai T
影响因子：
64.5
作者：
Kapoor A;Yao W;Ying H;Hua S;Liewen A;Wang Q;Zhong Y;Wu CJ;Sadanandam A;Hu B;Chang Q;Chu GC;Al-Khalil R;Jiang S;Xia H;Fletcher-Sananikone E;Lim C;Horwitz GI;Viale A;Pettazzoni P;Sanchez N;Wang H;Protopopov A;Zhang J;Heffernan T;Johnson RL;Chin L;Wang YA;Draetta G;DePinho RA
通讯作者：
DePinho RA
影响因子：
8
作者：
Azad, Taha;Nouri, Kazem;Yang, Xiaolong
通讯作者：
Yang, Xiaolong