PO.ET09.05 · 实验与分子治疗
(E,E)-bisantrene通过稳定c-MYC启动子区G-四链体沉默c-MYC表达
(E,E)-bisantrene silences c-MYC expression by stabilizing its promotor region G-quadruplex
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：G-四链体（G4）DNA和RNA是重要的非经典核酸二级结构，在许多细胞过程中发挥关键作用。它们调控多种癌基因的表达和翻译，包括细胞生长的主控调节因子MYC。Bisantrene是一种小分子抗癌药物，已在>1500例临床试验患者中被证明安全有效。本研究表征了(E,E)-bisantrene异构体对c-MYC启动子中一个G4区域的结合与稳定作用，以及对癌细胞中c-MYC表达的沉默。
方法：通过圆二色光谱确定(E,E)-bisantrene是否稳定c-MYC启动子G4结构，并使用表面等离子体共振测量结合亲和力。核磁共振波谱提供了配合物内结合相互作用的结构见解。分子动力学模拟对配合物的三维结构进行建模。在用(E,E)-bisantrene处理后，评估一系列癌细胞系中c-MYC基因表达的变化，并进行RNA-seq及通路分析。
结果：发现(E,E)-bisantrene可稳定c-MYC启动子G4区域，产生与其他G4配体（即pidnarulex和pyridostatin）相似的解链温度升高。NMR波谱和分子建模提示(E,E)-bisantrene以2:1化学计量比结合于顶部和底部G-四分体的平面表面。(E,E)-bisantrene在多种癌细胞系中强效抑制c-MYC表达。RNA-seq分析显示(E,E)-bisantrene还降低了启动子中含有G4区域的其他癌基因的表达，包括MET、TERT、VEGFA、ATF4和MDM2。RNA-Seq数据的通路分析显示其转录组特征与pidnarulex（一种处于早期临床开发的已知G4结合药物）相似。
结论：(E,E)-bisantrene结合并稳定c-MYC启动子区域内的G4结构，从而沉默c-MYC基因表达。这些研究支持将(E,E)-bisantrene作为一种新型G4靶向药物在MYC驱动肿瘤中进行临床评估。
查看英文原文 English abstract
Background: G-quadruplex (G4) DNA and RNA are important non-canonical nucleic acid secondary structures that play key roles in many cellular processes. They regulate the expression and translation of several oncogenes, including the master cell growth regulator, MYC. Bisantrene is a small-molecule anticancer agent that has been shown to be safe and effective in >1500 clinical trial patients. This study characterized the binding and stabilization of a G4 region in the c-MYC promoter by the (E,E)-bisantrene isomer and silencing of c-MYC expression in cancer cells.
Methods: Circular dichroism spectroscopy established if (E,E)-bisantrene stabilizes the c-MYC promotor G4 structure, with surface plasmon resonance used to measure the binding affinity. Nuclear magnetic resonance spectroscopy provided structural insights into the binding interactions within the complex. Molecular dynamics simulations modelled the 3-dimensional structure of the complex. Changes in c-MYC gene expression were assessed across a range of cancer cell lines after treatment with (E,E)-bisantrene and RNA-seq with pathway analysis was performed.
Results: (E,E)-bisantrene was found to stabilize the c-MYC promoter G4 region, producing similar increases in melting temperature to other G4 ligands (i.e., pidnarulex and pyridostatin). NMR spectroscopy and molecular modelling suggests (E,E)-bisantrene binds with a 2:1 stoichiometry to the planar surfaces of the top and bottom G-tetrads. (E,E)-bisantrene potently inhibited c-MYC expression in multiple cancer cell lines. RNA-seq analysis showed (E,E)-bisantrene also decreased expression of other oncogenes containing G4 regions in their promoters, including MET, TERT, VEGFA, ATF4 and MDM2 . Pathway analysis of the RNA-Seq data demonstrated a transcriptomic profile similar to pidnarulex, a known G4-binding drug in early-stage clinical development.
Conclusion: (E,E)-bisantrene binds to and stabilizes the G4 structure contained within the c-MYC promotor region, leading to silencing of c-MYC gene expression. These studies support clinical evaluation of (E,E)-bisantrene as a new G4-targeting drug in MYC-driven tumors.
利益披露 Disclosure
S. Sahni,
Race Oncology Employment, Stock Option.
E. Ryan,
Race Oncology Employment, Stock Option.
P. Cuthbertson,
Race Oncology Employment, Stock Option.
F. Ahmad,
Race Oncology Employment, Stock Option.
K. Curnow,
Race Oncology Employment, Stock Option.
N. Elsalamouny, None..
Q. Zhu, None.
H. Yu,
Race Oncology ).
J. Jang, None..
J. Dickerhoff, None.
D. Yang,
GIbson Oncology ).
E. Proctor, None..
M. Sanderson-Smith, None.
D. Tillett,
Race Oncology Employment, Stock, Stock Option.
M. Kelso,
Race Oncology Employment, Stock, Stock Option.