酪氨酸激酶FER促进卵巢癌扩散转移的功能及机制研究

卵巢癌是最令人绝望的妇科疾病，目前治疗手段非常有限，五年生存率极低。因此，确立分子靶标，研发新一代分子靶向药物，建立并完善卵巢癌的分子靶向治疗方案的需求非常迫切，并且具有不可估量的市场价值。申请人前期发表的工作证明酪氨酸激酶FER在调控卵巢癌细胞体内转移和药物耐受等方面均起着重要作用；当用RNAi技术敲减该基因以后，卵巢癌细胞在迁徙侵袭、扩散转移和抗药性等方面的能力都显著下降。在此基础上，申请者拟利用基因编辑（CRISPR-Cas9）技术，在细胞和动物层面上实现对FER基因的改造，使得改造过后的突变体只能使用特定的外源ATP类似物执行激酶功能，同时也只能被特定的小分子抑制剂灭活，从而实现对该激酶的体内精准实时调控。这套系统将进一步和卵巢癌遗传工程小鼠模型相结合，从而准确地评估酪氨酸激酶FER在卵巢癌发生、恶性发展以及药物拮抗中的作用，为验证其作为治疗卵巢癌的新靶点提供关键性的理论支持。

Ovarian cancer is the leading cause of death from gynecological malignancies. Due to limited therapeutic options, the overall survival from ovarian cancer has not improved significantly. Therefore, there is an urgent need and enormous marketing potential to develop and improve next-generation molecular targeted therapy in treating this deadly disease. Our previous study has demonstrated the indispensible role of tyrosine kinase FER in ovarian cancer metastasis and drug resistance, both of which attribute to high mortality rate of the disease. Loss of FER not only impaired the migration and invasion capability of ovarian cancer cell in vivo, but also enhanced the sensitivity of cancer cells to drug response. To further investigate the functional role of FER in ovarian tumorigenesis, in this current proposal, we plan to engineer FER protein by mutating its ATP-binding pocket, and screen 1) the mutant that can only use exogenous ATP analog to execute its kinase function; and 2) the small molecule inhibitor that can specifically antagonize the function of the mutant kinase. We will apply gene editing technology CRISPR-Cas9 to replace wild type FER kinase with this newly engineered mutant kinase on both ovarian cancer cell and animal levels. With these genetically engineered cell and animal models, along with currently available ovarian cancer mouse model, we could assess the important role of FER kinase in ovarian cancer oncogenesis, adverse progression and therapeutic resistance in an accurate and timely manner.