State Key Laboratory of Oncology in South China, Cancer Center, Collaborative Innovation Center of Cancer Medicine, Sun Yat-Sen University, Guangzhou, 510060, China.
Institute of Cancer Stem Cell, Cancer Center, Dalian Medical University, Dalian, 116023, China.
J Transl Med. 2023 Aug 18;21(1):553. doi: 10.1186/s12967-023-04396-w.
Triple-negative breast cancer (TNBC) is one of the most aggressive human cancers and has poor prognosis. Approximately 80% of TNBC cases belong to the molecular basal-like subtype, which can be exploited therapeutically by inducing differentiation. However, the strategies for inducing the differentiation of TNBC remain underexplored.
A three-dimensional (3D) morphological screening model based on a natural compound library was used to identify possible candidate compounds that can induce TNBC cell differentiation. The efficacy of rutaecarpine was verified using assays: RT-qPCR, RNA-seq, flow cytometry, immunofluorescence, SCENITH and label-free LC-MS/MS. The direct targets of rutaecarpine were identified through drug affinity responsive target stability (DARTS) assay. A xenograft mice model was also constructed to confirm the effect of rutaecarpine in vivo.
We identified that rutaecarpine, an indolopyridoquinazolinone, induces luminal differentiation of basal TNBC cells in both 3D spheroids and in vivo mice models. Mechanistically, rutaecarpine treatment leads to global metabolic stress and elevated ROS in 3D cultured TNBC cells. Moreover, NAC, a scavenger of ROS, impedes rutaecarpine-induced differentiation of TNBC cells in 3D culture. Finally, we identified fumarate hydratase (FH) as the direct interacting target of rutaecarpine. The inhibition of FH and the knockdown of FH consistently induced the differentiation of TNBC cells in 3D culture.
Our results provide a platform for differentiation therapy drug discovery using 3D culture models and identify rutaecarpine as a potential compound for TNBC treatment.
三阴性乳腺癌（TNBC）是最具侵袭性的人类癌症之一，预后不良。大约 80%的 TNBC 病例属于分子基底样亚型，可以通过诱导分化进行治疗。然而，诱导 TNBC 分化的策略仍未得到充分探索。
使用基于天然化合物库的三维（3D）形态筛选模型来鉴定可能的候选化合物，这些化合物可以诱导 TNBC 细胞分化。使用 RT-qPCR、RNA-seq、流式细胞术、免疫荧光、SCENITH 和无标记 LC-MS/MS 等检测方法验证了荷叶碱的功效。通过药物亲和反应靶标稳定性（DARTS）测定法鉴定了荷叶碱的直接靶标。还构建了异种移植小鼠模型以确认荷叶碱在体内的作用。
我们发现，荷叶碱，一种吲哚并吡啶喹唑啉酮，可诱导基底 TNBC 细胞在 3D 球体和体内小鼠模型中发生管腔分化。在机制上，荷叶碱处理导致 3D 培养的 TNBC 细胞中产生全局性代谢应激和升高的 ROS。此外，ROS 的清除剂 NAC 可阻止荷叶碱诱导的 3D 培养中的 TNBC 细胞分化。最后，我们确定延胡索酸水合酶（FH）为荷叶碱的直接作用靶标。FH 的抑制和 FH 的敲低一致诱导了 3D 培养中的 TNBC 细胞分化。
我们的研究结果为使用 3D 培养模型进行分化治疗药物发现提供了一个平台，并确定荷叶碱是一种潜在的 TNBC 治疗化合物。
