Integrating Diagnostics with Therapeutic Strategies in Chronic Myeloid Leukemia

描述（由申请人提供）：慢性髓性白血病（CML）提供了一种独特的疾病模型，可以将转化方法应用于疾病进展、治疗反应和疾病复发的生物学研究。尽管BCR-ABL的功能已被很好地描述，但与疾病进展有关的事件在很大程度上仍然未知。在过去的10年里，酪氨酸激酶抑制剂如甲磺酸伊马替尼（IM）极大地改变了CML的治疗。然而，尽管在大多数早期慢性期（CP）患者中有良好的反应，但约20%的CP患者未能接受IM治疗。这一数字在晚期CP患者中更高，而晚期疾病或对IM具有耐药性的疾病的预后明显较差。因此，显然需要早期识别有疾病进展或治疗耐药风险的患者。提出的转化研究依赖于识别患者的危险因素，然后在独立患者中进行验证，最终目标是将这些发现应用于临床环境中的患者护理。这些研究的成功依赖于在足够多的患者中测试发现的能力。我们正在积极参与一些CML的临床试验，包括SWOG S0325，该试验将标准剂量IM与高剂量IM和达沙替尼进行比较。在Specific Aim 1中，先前在微阵列研究中发现的与CML疾病进展和治疗耐药相关的诊断基因表达候选者将在400个独立患者样本中使用高通量定量RT-PCR （QPCR）进行验证。研究将扩大到使用高通量QPCR分析所有目前已知的miRNAs来鉴定与CML进展和治疗耐药性相关的microRNAs （miRNAs）。在Specific Aim 2中，这些诊断研究将与接受酪氨酸激酶抑制剂（TKIs）的患者的强化监测研究相结合。这些研究将包括早期检测和监测T315I的发展动力学，T315I是一种对所有目前批准的TKIs具有抗性的突变。增强的QPCR监测bcr-abl患者保持最佳的分子反应也将进行使用新型纳米流体平台，提高灵敏度通过样品划分为多达10,000个独立的反应。T315I点突变研究的目的是确定早期T315I突变检测是否可以用于指导治疗选择；在最佳分子反应患者中加强bcr-abl监测的目标是确定动力学特征，可能潜在地确定可以停止TKI治疗的患者。最后，在Specific Aim 3中，我们将使用慢病毒载体基因沉默和表达技术研究PRAME和GLI2在细胞系和原代细胞的疾病进展和治疗耐药性中的作用。主要目标将是确定参与进展和耐药的新途径，这些途径可以通过现有疗法或新策略来靶向。这些综合生物学和临床研究的未来主要临床应用是确定可用于确定个体患者基于风险的治疗管理的概况。

DESCRIPTION (provided by applicant): Chronic myeloid leukemia (CML) provides a unique disease model in which to apply a translational approach to the study of the biology of disease progression, therapeutic response, and disease relapse. Although the function of BCR-ABL is well described, events involved in disease progression remain largely unknown. Over the last 10 years, tyrosine kinase inhibitors such as imatinib mesylate (IM) have dramatically altered CML treatment. Yet despite the excellent responses in most early chronic phase (CP) patients, ~20% of CP patients fail IM therapy. This number is higher for late CP patients and outcomes for advanced disease or disease resistant to IM are significantly poorer. Thus, a clear need exists for the early identification of patients at risk for disease progression or therapeutic resistance. The proposed translational studies rely on the identification of risk factors in patients, followed by validation in independent patients, with the ultimate goal of applying these finding to patient care in the clinical setting. The success of these investigations relies on the ability to test the findings in a sufficiently large number of patients. We are actively participating in a number of clinical trials for CML, including SWOG S0325, which compares standard dose IM to high dose IM and to dasatinib. In Specific Aim 1 diagnostic gene expression candidates associated with CML disease progression and therapy resistance that were previously identified in microarray studies will be validated using high throughput quantitative RT-PCR (QPCR) in 400 independent patient samples. Investigations will be expanded to identify microRNAs (miRNAs) associated with CML progression and therapy resistance using high throughput QPCR profiling for all currently known miRNAs. In Specific Aim 2 these diagnostic studies will be integrated with enhanced monitoring studies in patients receiving tyrosine kinase inhibitors (TKIs). These investigations will include early detection and monitoring of the kinetics of T315I development, a mutation resistant to all currently approved TKIs. Enhanced QPCR monitoring of bcr-abl in patients maintaining the best molecular responses will also be performed using a novel nanofluidic platform that enhances sensitivity through sample partitioning into up to 10,000 independent reactions. The goal of the T315I point mutation studies is to determine whether early T315I mutation detection can be used to guide therapeutic choices; the goal of enhanced bcr-abl monitoring in the best molecular response patients is to determine a kinetic profile that may potentially identify patients in whom TKI therapy can be stopped. Lastly, in Specific Aim 3 we will examine PRAME and GLI2 for their role in disease progression and therapy resistance in cell lines and primary cells using lentiviral vector-based gene silencing and expression techniques. A main goal will be to identify new pathways involved in progression and resistance that can be targeted either by existing therapies or by novel strategies. A major future clinical application of these integrated biological and clinical studies is to identify profiles that can be used to determine risk-based treatment management in individual patients.