Examination of FLT3 Mutations in Acute Myeloid Leukemia

描述（由申请人提供）：Stirewalt博士的长期目标是 为AML患者开发治疗方法，降低发病率， 提高生存率。 在接下来的五年里，斯蒂雷沃特博士将进行预- 研究FLT 3突变对人类免疫功能的影响的临床研究。 急性髓细胞白血病 具体目标I：检查FLT 3突变的临床意义 既往未经治疗的成人AML患者。 的预后意义 成人AML患者的FLT 3突变仍不确定。 具体目标I 前瞻性研究FLT 3突变在成人中的临床意义 AML患者被分配到SWOG S 0106，这是一项III期试验， 剂量诱导和高剂量Ara-C巩固，随后随机分配至 观察或Mylotarg。 AML患者将接受FLT 3突变筛查 使用灵敏的PCR/SSCP分析。 突变数据将与 临床结果。 我们假设FLT 3 ITD将是一个独立的 成人AML患者的预后不良因素。 特定目的II：检查FLT 3 ITD PCR检测试剂盒预测 临床复发 大约90%的FLT 3 ITD发生在 没有MRD的分子标记，我们已经开发了灵敏的PCR检测方法， 对于可用作MRD检测的FLT 3 ITD。 具体目标二审查了 FLT 3 ITD的MRD检测预测接受以下治疗的患者临床复发的能力 SWOG S0106。 将在所有FLT 3中进行FLT 3 ITD的MRD测定 治疗期间特定时间点的阳性患者和结果 将与临床结果相关。 我们假设MRD检测 使用FLT 3 ITD将预测临床复发。 具体目标III：检查参与激活的分子效应物 突变的FLT 3受体，并开发技术来抵消 突变的FLT受体。 来自转染FLT 3 ITD的鼠细胞的数据 表明FLT 3 ITD导致FLT 3受体的激活， 被非特异性酪氨酸激酶抑制剂抑制。 我们将转化 将基因工程化的FL T3突变导入鼠细胞中。 的影响 这些FLT 3突变对受体、MAP激酶、STAT 3&5、c- Jun和全球基因表达将被检查。 我们还将确定 如果抗体直接针对FL T3受体，新的酪氨酸激酶 抑制剂和/或非受体酪氨酸激酶抑制剂可以改变 FLT 3突变的影响。 使用患者AML样品的平行实验 还将进行FLT 3 ITD。 我们假设FLT 3 ITD将 受体的组成性激活可被阻断， 酪氨酸激酶抑制剂。 除了这项研究计划外，Stirewalt博士还制定了一项 教育计划，包括具体的培训目标，教学 课程，以及他的顾问的正式审查。 该教育计划将 加强他在临床试验设计和方法学方面的背景。 在一起， 研究建议和教育计划为Dc Stirewalt提供了 成为一名独立的临床研究者。

DESCRIPTION (provided by applicant): Dr. Stirewalt's long-term objective is to develop treatments for patients with AML that will reduce morbidity and improve survival. Over the next five years, Dr. Stirewalt will perform pre- clinical and clinical studies that examine the effects of FLT3 mutations in AML. Specific Aim I: Examine the clinical significance of FLT3 mutations in previously untreated adult patients with AML. The prognostic significance of FLT3 mutations in adults with AML remains uncertain. Specific Aim I prospectively examines the clinical significance of FLT3 mutations in adult AML patients assigned to SWOG SOl06, a phase III trial consisting of standard dose induction and high dose Ara-C consolidation followed by randomization to observation or Mylotarg. AML patients will be screened for FLT3 mutations using sensitive PCR/SSCP assays. Mutation data will be correlated with clinical outcomes. We hypothesize that FLT3 ITDs will be an independent negative prognostic factor in adult patients with AML. Specific Aim II: Examine the ability of PCR assays for FLT3 ITDs to predict clinical relapse. Approximately 90 percent of FLT3 ITDs occur in patients without a molecular marker for MRD, and we have developed sensitive PCR assays for FLT3 ITDs that may be used as tests for MRD. Specific Aim II examines the ability of MRD assays for FLT3 ITDs to predict clinical relapse in patients on SWOG S0106. The MRD assays for FLT3 ITDs will be performed in all FLT3 positive patients at specific time points during their treatment and results will be correlated with clinical outcomes. We hypothesize that MRD assays using FLT3 ITDs will be predictive of clinical relapse. Specific Aim III: Examine the molecular effectors involved in the activation of mutated FLT3 receptors and develop techniques to counteract the effects of mutated FLT receptors. Data from murine cells with transfected FLT3 ITDs suggest that FLT3 ITDs result in activation of the FL T3 receptors that can be inhibited by nonspecific tyrosine kinase inhibitors. We will transfact genetically engineered FL T3 mutations into murine cells. The effects of these FLT3 mutations on activation of the receptor, MAP kinase, STAT 3&5, c- Jun, and global genetic expression will be examined. We will also determine if antibodies directed against the FL T3 receptor, novel tyrosine kinese inhibitors, and/or non-receptor tyrosine kinase inhibitors can alter the effects of FLT3 mutations. Parallel experiments using patient AML samples with FLT3 ITDs will also be performed. We hypothesize that FLT3 ITDs will premote constitutive activation of the receptor that can be blocked by tyrosine kinase inhibitors. In addition to the research proposal, Dr. Stirewalt has developed an educational program that includes specific training objectives, didactic courses, and formal reviews by his advisors. This educational program will strengthen his background in clinical trial design and methodology. Together, the research proposal and educational program provide Dc Stirewalt with the skills to become an independent clinical investigator.