同源异型盒转录因子HOXB13诱导肺腺癌耐药和转移的分子机制及临床诊断意义

Resistance to chemotherapy and metastasis of lung adenocarcinomas are the main causes of therapeutic failure in clinical patients. In our previous works we found that transcriptional factor HOXB13 is associated with drug resistance, clinical grading and prognosis of lung adenocarcinoma. HOXB13 plays an important role in embryonic development and tumorigenesis, but its role in lung cancer progression has not been reported. This grant proposal will identify the relationship between HOXB13 and lung adenocarcinoma progression, outcomes and resistance to chemotherapy by analyzing enlarged amount of clinical samples and also TCGA big datasets. We will clarify whether HOXB13 promotes drug resistance and metastasis of lung adenocarcinoma cells and induces the subpopulation of lung cancer stem cells through in vivo and in vitro functional experiments. Then we will systematically uncover the molecular mechanisms underlying HOXB13 regulation on drug resistance and metastasis in lung adenocarcinoma cells at the whole genome level by identifying the downstream target genes of HOXB13. Then we will identify the interacting proteins of HOXB13 in lung cancer cells. HOXB13 and the combination of its target genes will be examined for their usefulness in predicting the progression, drug resistance and prognosis of lung adenocarcinomas from biopsies of the patients. This study proposed for the first time that HOXB13 is associated with the resistance to chemotherapy. We seek to investigate the advantage and necessity of combined application of HOXB13 and its target genes to evaluate the therapeutic effects and prognosis for lung adenocarcinoma patients. This investigation anticipates to provide a new group of candidate genes for evaluation of drug resistance, metastasis and prognosis for lung adenocarcinoma patients.

肺腺癌的耐药和转移是临床治疗失败的主要原因。我们前期发现转录因子HOXB13与肺腺癌耐药、临床分级和预后生存相关。HOXB13在胚胎发育和肿瘤发生中起重要作用，但其与肺癌之间的关系尚无报道。本申请将结合临床病例检测及TCGA大数据库，明确HOXB13与肺腺癌化疗耐药、进展和预后之间的关系；通过体内、外功能实验明确HOXB13促进肺腺癌细胞耐药、转移及诱导肺癌干细胞亚群的功能；在全基因组范围内揭示HOXB13调控耐药和转移的下游靶基因，揭示HOXB13的相互作用蛋白，系统阐述HOXB13促进肺腺癌耐药与转移的分子机制。将结合肺腺癌病人穿刺活检，通过IHC和基因表达的定量检测，采用HOXB13及其靶基因组联合分析预测肺腺癌的耐药和预后。本研究首次提出了HOXB13与化疗耐药有关，并将探讨HOXB13及其靶基因联合应用的优越性及必要性，为肺腺癌的化疗耐药、转移倾向和预后评价提供新的判断方案。

肺癌的转移和耐药是其治疗失败、生存期短的主要原因，尤其是继发耐药使本来有效的治疗变得失败。我们首先在肺腺癌耐药患者组织中发现同源异形盒转录因子HOXB13表达异常上调，并且HOXB13表达的增高对应着患者癌组织更差的临床分期和更不良的预后。继而研究组对于HOXB13的生物学功能进行了详细解析，发现高表达HOXB13的细胞具有更强大的增殖能力和运动侵袭能力，并具有更强的耐药性。这些细胞到了动物体内表现出更早的成瘤能力及强大的局部侵袭和远隔器官转移能力。课题组在全基因组范围内检测了HOXB13可能的下游靶基因，并对每个靶基因进行了逐一验证，确定了其中ABCG1、EZH2、Slug及HOXB13自身是受HOXB13 直接靶向并上调的下游分子。ABC转运体ABCG1和重要表观遗传调控因子EZH2与启动子结合而产生耐药性；而EZH2和Slug是HOXB13促迁移侵袭的调控分子；同时HOXB13通过自身正反馈调节形成更强大的调节环路使其生物学功能再度被放大。更为重要的是研究组发现顺铂在癌细胞和患者来源的移植物（PDX）中能够显著诱导HOXB13的表达上调。因此，该发现是化疗继发耐药的关键机制，也是控制肺腺癌继发耐药的有效靶点。联合应用HOXB13及其靶向基因ABCG1和EZH2比单分子应用获得更有效的预测患者对基于铂类药物治疗的敏感性、治疗效果和患者预后，从而可作为临床监测肺腺癌患者耐药性和预测预后的备选指标。本研究已经申请国家专利局专利。上述发现得到Theranostics资深编辑和论文审稿人的认同，论文同时获得高度评价。该研究的意义在于HOXB13及其靶向基因（包括ABCG1、EZH2和IL6）的联合应用将预测患者对基于铂类药物治疗的敏感性，从而可作为监测肺腺癌患者耐药性和预测预后的指标。在该基金的支持下我们研究组发表10篇论文（其中综述2篇，Q1区文章7篇，影响因子大于8的4篇）、申请其它基金资助7项（共计300万）、培养博士6名，硕士3名，参加国内外会议，获得若干奖项。总之，在该基金资助下，本人超额完成预计研究计划。