TAS::75 0849::TAS SBIR TOPIC 277 - DEVELOPMENT OF A CLINICAL DIAGNOSTIC ASSAY FO

激酶抑制剂使癌症治疗发生了革命性的变化，但它们的使用导致了耐药性，促使了针对耐药肿瘤的后续抑制剂的开发。间变性淋巴瘤激酶(ALK)是由斯蒂芬·莫里斯博士发现的(《洞察遗传学》是这些研究的合作者)。ALK融合基因和激活点突变会导致几种癌症，多家制药公司正在开发ALK抑制剂。一种抑制剂PF-02341066(Crizotinib，辉瑞)已经进入第三阶段试验，可能会在2011年获得FDA的批准。莫里斯小组已经确定了大量的ALK激酶结构域突变，这些突变赋予了高水平的PF-02341066耐药性，这已经在患者中得到了报道。需要临床基因测试来识别耐药突变，以指导对第一代疗法耐药的患者开发和使用下一代ALK抑制剂。本文提出的工作将开发一种针对ALK抑制剂耐药突变的临床诊断方法，以满足这一未得到满足的需求。目的1.建立和应用等位基因特异的聚合酶链式反应平台检测所有已知的ALK抑制剂耐药突变的基因诊断方法。目的：验证该检测方法对耐碱性磷酸酶抑制剂的细胞株和对PF-02341066耐药的患者的肺癌标本进行检测的能力。

Kinase inhibitors have revolutionized cancer treatment but their use has led to drug resistance, prompting development of follow-on inhibitors for resistant tumors. Anaplastic lymphoma kinase (ALK) was discovered by Dr. Stephan Morris (Insight Genetics¿ collaborator on these studies). ALK fusion genes and activating point mutations cause several cancers, and multiple pharmaceutical firms are developing ALK inhibitors. One inhibitor ¿ PF-02341066 (crizotinib, Pfizer) ¿ has entered Phase III trials and will likely receive FDA approval in 2011. The Morris group has identified a large number of ALK kinase domain mutations that confer high-level PF-02341066 resistance, which has already been reported in patients. A clinical genetic test to identify inhibitor-resistance mutations is needed to guide the development and use of next-generation ALK inhibitors for patients resistant to 1st-generation therapy. The work proposed herein will develop a clinical diagnostic assay for ALK inhibitor-resistance mutations to address this unmet need. Objective 1. Development and application of a genetic diagnostic assay to detect all known ALK inhibitor-resistance mutations using an allele-specific PCR platform. Objective 2: Demonstration and validation of the ability of the test to identify ALK mutations from both cell lines with ALK inhibitor resistance and re-biopsied lung cancer specimens from patients who developed PF-02341066 resistance.