三维CRISPR筛选揭示了与抗血管生成治疗的表观遗传相互作用

血管生成是癌症、眼睛和心血管疾病的发展、生理和发病机制的基础。抗血管生成治疗(AAT)抑制异常血管生成已成功应用于临床治疗肿瘤和眼部疾病。然而，对 AAT 的抗性不可避免地发生了，其分子基础仍然知之甚少。在这里，我们揭示了血管内皮细胞(EC)对广泛使用的 AAT 贝伐单抗反应的分子修饰剂，通过使用基于三维微载体的细胞培养和 CRISPR-Cas9进行混合基因筛选。表观遗传学阅读器 BET 家族的 BRD2/3/4蛋白的功能抑制显示了阻断 VEGFA 对 EC 存活和/或增殖的意外缓解作用。

此外，转录组学和通路分析揭示了表观遗传调控和抗血管生成之间的相互作用，这可能通过细胞周期调节因子 CDC25B 磷酸酶影响内皮细胞的染色体结构和活性。总的来说，我们的研究结果促进人们对 EC 对 VEGFA 封锁反应的表观遗传调控的了解，并可能促进优质生物标志物和克服 AAT 抗性的策略的发展。

Angiogenesis underlies development, physiology and pathogenesis of cancer, eye and cardiovascular diseases. Inhibiting aberrant angiogenesis using anti-angiogenic therapy (AAT) has been successful in the clinical treatment of cancer and eye diseases. However, resistance to AAT inevitably occurs and its molecular basis remains poorly understood. Here, we uncover molecular modifiers of the blood endothelial cell (EC) response to a widely used AAT bevacizumab by performing a pooled genetic screen using three-dimensional microcarrier-based cell culture and CRISPR–Cas9. Functional inhibition of the epigenetic reader BET family of proteins BRD2/3/4 shows unexpected mitigating effects on EC survival and/or proliferation upon VEGFA blockade. Moreover, transcriptomic and pathway analyses reveal an interaction between epigenetic regulation and anti-angiogenesis, which may affect chromosomal structure and activity in ECs via the cell cycle regulator CDC25B phosphatase. Collectively, our findings provide insight into epigenetic regulation of the EC response to VEGFA blockade and may facilitate development of quality biomarkers and strategies for overcoming resistance to AAT.

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