靶向治疗在乳腺癌中取得突破，但肿瘤干细胞导致的耐药是制约疗效的瓶颈。我们前期发现细胞周期激酶AURKA核转位激活FOXM1/Myc，促进乳腺癌干性，抵抗激酶靶向治疗(Oncogene 2017；Nat Commun 2016)。申请人筛选AURKA胞核互作蛋白发现其与染色质重构蛋白RAD21/p300结合；ChIP-seq发现其基因组结合位点多伴随H3K27ac修饰；染色质构象捕获实验证实其参与了染色质构象重塑。上述结果提示AURKA介导染色质重塑，促进（超级）增强子形成，增强干性基因表达，进而导致耐药。本项目拟在已建立的耐药和核转位模型中，明确AURKA调控染色质三维重构的共作用因子，揭示其复合物形成及作用的机制；阐明其促干性关键靶基因；探寻靶向AURKA核功能的干预靶点及临床意义。本项目是靶向AURKA抗癌领域的远瞻课题，有望揭示AURKA全新的促癌机制，为靶向乳腺癌干细胞提供新靶点。

Molecularly targeted cancer therapy has achieved tremendous advances in the past decade. However, recent studies suggest that cancer stem cells (CSCs) lead to therapy resistance, tumor progression, and recurrence. Based on AURKA-targeted therapy resistance model and AURKA nuclear translocation model, we have found that oncogenic kinase AURKA translocated into nucleus and lead to drug resistance through promoting cancer stemness (Oncogene 2017; Nat Commun 2016). Recently, we performed stable isotope labeling with amino acids in cell culture (SILAC) assays and chromatin immunoprecipitation-sequencing (ChIP-seq) assays to get the interacting proteins and genomic DNA binding sites of nuclear AURKA, respectively. We found that AURKA bind to genome organization protein RAD21/p300 and its binding sites frequently occurred H3K27ac modification. We further performed chromatin conformation capture assay (3C) and found the space interaction of AURKA binding sites. These results suggest that AURKA translocated to nucleus and remodeled three-dimensional genome organization under chemotherapy or AURKA-targeted therapy, thus promoting the formation of (super) enhancer, enhancing cancer stemness, and leading to drug resistance. This project aims to exploit molecular mechanisms of AURKA-dependent 3D genome organization in BCSCs, determine co-factors, illustrate the key target genes, and investigate the clinical significance of AURKA nuclear translocation. Therefore, this project will provide novel target for cancer stemness, and present new rationale for oncogenic kinase targeting.