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

激酶抑制剂已经彻底改变了癌症治疗，但它们的使用导致了耐药性，促进了耐药肿瘤后续抑制剂的开发。间变性淋巴瘤激酶（ALK）是由Stephan Morris博士（Insight Genetics的合作者）发现的。ALK融合基因和激活点突变导致多种癌症，多家制药公司正在开发ALK抑制剂。一种抑制剂PF-02341066（克唑替尼，辉瑞）已进入III期试验，并可能在2011年获得FDA批准。Morris研究小组已经确定了大量ALK激酶结构域突变，这些突变可导致高水平PF-02341066耐药，这在患者中已有报告。需要进行临床遗传学检测以确定耐药突变，以指导下一代ALK抑制剂的开发和使用，用于对第一代治疗耐药的患者。本文提出的工作将开发一种ALK耐药突变的临床诊断测定法，以解决这一未满足的需求。目的1.开发和应用一种遗传诊断试验，使用等位基因特异性PCR平台检测所有已知的ALK靶标耐药突变。目标二：证明和验证该检测方法能够从ALK抑制剂耐药细胞系和PF-02341066耐药患者的再活检肺癌标本中鉴别ALK突变。

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.