芳香烃受体核转位蛋白样（ARNTL/BMAL1）通过调节血管内皮生长因子 A 与结直肠癌细胞对贝伐珠单抗的耐药性相关。
Aryl hydrocarbon receptor nuclear translocator-like (ARNTL/BMAL1) is associated with bevacizumab resistance in colorectal cancer via regulation of vascular endothelial growth factor A.
机构信息
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany.
Division of Medical Oncology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, CA, United States; Unit of Medical Oncology 1, Clinical and Experimental Oncology Department, Veneto Institute of Oncology IOV - IRCCS, Padua, Italy.
出版信息
EBioMedicine. 2019 Jul;45:139-154. doi: 10.1016/j.ebiom.2019.07.004. Epub 2019 Jul 9.
BACKGROUND
The identification of new biomarkers and the development of novel, targetable contexts of vulnerability are of urgent clinical need in drug-resistant metastatic colorectal cancer (mCRC). Aryl-Hydrocarbon-Receptor-Nuclear-Translocator-Like (ARNTL/BMAL1) is a circadian clock-regulated transcription factor promoting expression of genes involved in angiogenesis and tumour progression. We hypothesised that BMAL1 increases expression of the vascular endothelial growth factor A VEGFA gene and, thereby, confers resistance to anti-angiogenic therapy with bevacizumab (Beva), a clinically used antibody for neutralization of VEGFA.
METHODS
PCR and immunohistochemistry were employed to assess BMAL1 expression in mice (C57BL/6 J ; BALB/c xenografts) and CRC patients under combination chemotherapy with Beva. BMAL1 single nucleotide gene polymorphisms (SNPs) were analysed by DNA-microarray in clinical samples. BMAL1 functions were studied in human CRC cell lines using colorimetric growth, DNA-binding and reporter assays.
FINDINGS
In murine CRCs, high BMAL1 expression correlated with poor preclinical response to Beva treatment. In CRC patients' tumours (n = 74), high BMAL1 expression was associated with clinical non-response to combination chemotherapy with Beva (*p = .0061) and reduced progression-free survival (PFS) [*p = .0223, Hazard Ratio (HR) = 1.69]. BMAL1 SNPs also correlated with shorter PFS (rs7396943, rs7938307, rs2279287) and overall survival (OS) [rs11022780, *p = .014, HR = 1.61]. Mechanistically, Nuclear-Receptor-Subfamily-1-Group-D-Member-1 (NR1D1/REVERBA) bound a - 672 bp Retinoic-Acid-Receptor-Related-Orphan-Receptor-Alpha-responsive-element (RORE) adjacent to a BMAL1 DNA-binding motif (E-box) in the VEGFA gene promoter, resulting in increased VEGFA synthesis and proliferation of human CRC cell lines.
INTERPRETATION
BMAL1 was associated with Beva resistance in CRC. Inhibition of REVERBA-BMAL1 signalling may prevent resistance to anti-angiogenic therapy. FUND: This work was in part supported by the European Commission Seventh Framework Programme (Contract No. 278981 [ANGIOPREDICT]).
背景
在耐药转移性结直肠癌（mCRC）中，鉴定新的生物标志物和开发新的、可靶向的脆弱性靶点是迫切的临床需求。芳香烃受体核转位蛋白样（ARNTL/BMAL1）是一种昼夜节律调节转录因子，可促进参与血管生成和肿瘤进展的基因表达。我们假设 BMAL1 增加血管内皮生长因子 A（VEGFA）基因的表达，从而对贝伐单抗（Beva）的抗血管生成治疗产生耐药性，Beva 是一种用于中和 VEGFA 的临床应用抗体。
方法
采用 PCR 和免疫组织化学方法检测 C57BL/6J；BALB/c 异种移植小鼠和 CRC 患者在联合贝伐单抗化疗下的 BMAL1 表达。通过 DNA 微阵列分析临床样本中的 BMAL1 单核苷酸基因多态性（SNP）。使用比色生长、DNA 结合和报告基因测定研究人 CRC 细胞系中的 BMAL1 功能。
发现
在小鼠 CRC 中，高 BMAL1 表达与对贝伐单抗治疗的临床前反应不佳相关。在 CRC 患者的肿瘤（n=74）中，高 BMAL1 表达与联合贝伐单抗化疗的临床无反应相关（*p=0.0061），并降低无进展生存期（PFS）[*p=0.0223，风险比（HR）=1.69]。BMAL1 SNP 也与较短的 PFS（rs7396943、rs7938307、rs2279287）和总生存期（OS）相关（rs11022780，*p=0.014，HR=1.61）。从机制上讲，核受体亚家族 1 组 D 成员 1（NR1D1/REVERBA）与 VEGFA 基因启动子中靠近 BMAL1 DNA 结合基序（E 盒）的-672bp 视黄酸受体相关孤儿受体-α反应元件（RORE）结合，导致 VEGFA 合成增加和人 CRC 细胞系增殖。
解释
BMAL1 与 CRC 中的 Beva 耐药性相关。抑制 REVERBA-BMAL1 信号可能会防止对抗血管生成治疗的耐药性。
资金
这项工作部分得到了欧盟第七框架计划（合同号 278981[ANGIOPREDICT]）的支持。
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