Metastasis-associated MCL1 and P16 copy number alterations dictate resistance to vemurafenib in a BRAFV600E patient-derived papillary thyroid carcinoma preclinical model.
Metastasis-associated MCL1 and P16 copy number alterations dictate resistance to vemurafenib in a BRAFV600E patient-derived papillary thyroid carcinoma preclinical model.
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DOI：
10.18632/oncotarget.6442
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发表时间：
2015-12-15
期刊：
影响因子：
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通讯作者：
Nucera C
中科院分区：
文献类型：
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作者：
Duquette M;Sadow PM;Husain A;Sims JN;Antonello ZA;Fischer AH;Song C;Castellanos-Rizaldos E;Makrigiorgos GM;Kurebayashi J;Nose V;Van Hummelen P;Bronson RT;Vinco M;Giordano TJ;Dias-Santagata D;Pandolfi PP;Nucera C
BRAFV 600 E突变在许多肿瘤中发挥重要的致癌功能，包括甲状腺乳头状癌（PTC）。虽然BRAFV 600 E抑制剂是可用的，但经常观察到缺乏反应。为了研究对突变型BRAFV 600 E选择性抑制剂维罗非尼的内在抗性的机制，我们建立了人转移性/复发性BRAFV 600 E-PTC、甲状腺内BRAFV 600 E-PTC和正常甲状腺（NT）的短期原代细胞培养物。我们还产生了人BRAFV 600 E-PTC原位小鼠的早期干预模型。我们发现，转移性BRAFV 600 E-PTC细胞引发旁分泌信号，触发周细胞，血液内皮细胞和淋巴管内皮细胞的迁移相比，BRAFWT-PTC细胞，并显示出更高的侵袭率。我们进一步表明，与媒介物治疗相比，维罗非尼治疗显著抑制非转移性BRAFV 600 E-PTC细胞中的这些异常功能，但在转移性BRAFV 600 E-PTC细胞中较少。这些结果与肿瘤微环境相关的促转移分子下调的相似倍数一致，在BRAFWT-PTC和NT细胞中没有影响。我们的早期干预临床前试验表明，vemurafenib延迟原位BRAFWT/V600 E-PTC小鼠的肿瘤生长。重要的是，我们鉴定了转移性BRAFV 600 E-PTC细胞中MCL 1（染色体1 q）的高拷贝数增加和CDKN 2A（P16，染色体9 p）的丢失，这与对维罗非尼治疗的抗性相关。重要的是，我们证明了与单一药物治疗相比，维罗非尼与BCL 2/MCL 1抑制剂的组合治疗增加了转移性BRAFV 600 E-PTC细胞死亡并改善了对维罗非尼治疗的反应。总之，短期PTC和NT培养为评估PTC患者的治疗反应提供了预测模型。我们的PTC临床前模型表明，联合靶向治疗可能是转移性和难治性BRAFV 600 E阳性PTC的重要治疗策略。
BRAFV600E mutation exerts an essential oncogenic function in many tumors, including papillary thyroid carcinoma (PTC). Although BRAFV600E inhibitors are available, lack of response has been frequently observed. To study the mechanism underlying intrinsic resistance to the mutant BRAFV600E selective inhibitor vemurafenib, we established short-term primary cell cultures of human metastatic/recurrent BRAFV600E-PTC, intrathyroidal BRAFV600E-PTC, and normal thyroid (NT). We also generated an early intervention model of human BRAFV600E-PTC orthotopic mouse. We find that metastatic BRAFV600E-PTC cells elicit paracrine-signaling which trigger migration of pericytes, blood endothelial cells and lymphatic endothelial cells as compared to BRAFWT-PTC cells, and show a higher rate of invasion. We further show that vemurafenib therapy significantly suppresses these aberrant functions in non-metastatic BRAFV600E-PTC cells but lesser in metastatic BRAFV600E-PTC cells as compared to vehicle treatment. These results concur with similar folds of down-regulation of tumor microenvironment–associated pro-metastatic molecules, with no effects in BRAFWT-PTC and NT cells. Our early intervention preclinical trial shows that vemurafenib delays tumor growth in the orthotopic BRAFWT/V600E-PTC mice. Importantly, we identify high copy number gain of MCL1 (chromosome 1q) and loss of CDKN2A (P16, chromosome 9p) in metastatic BRAFV600E-PTC cells which are associated with resistance to vemurafenib treatment. Critically, we demonstrate that combined vemurafenib therapy with BCL2/MCL1 inhibitor increases metastatic BRAFV600E-PTC cell death and ameliorates response to vemurafenib treatment as compared to single agent treatment. In conclusion, short-term PTC and NT cultures offer a predictive model for evaluating therapeutic response in patients with PTC. Our PTC pre-clinical model suggests that combined targeted therapy might be an important therapeutic strategy for metastatic and refractory BRAFV600E-positive PTC.