Examination of FLT3 Mutations in Acute Myeloid Leukemia

描述(申请人提供)：Stirewalt博士的长期目标是 为急性髓细胞白血病患者开发治疗方法，以降低发病率和 提高存活率。在接下来的五年里，斯特雷沃特博士将在 临床和临床研究检验了Flt3突变对 AML。 特定目的I：检测人外周血中Flt3突变的临床意义 以前未经治疗的成人AML患者。血管紧张素转换酶抑制剂的预后意义 成人AML患者中的Flt3突变仍不确定。具体目标一 前瞻性研究成人Flt3基因突变的临床意义 AML患者被分配到SWOG SO106，这是一项由标准试验组成的III期试验 剂量诱导和大剂量阿糖胞苷巩固随机化治疗 观察或Mylotarg。将对AML患者进行Flt3突变筛查 采用敏感的聚合酶链式反应/单链构象多态性分析。突变数据将与 临床结果。我们假设Flt3ITD将是一个独立的 成人急性髓系白血病患者的阴性预后因素。 特定目标II：检验聚合酶链式反应检测Flt3ITDS的能力 临床复发。大约90%的FLT3ITD发生在患者 没有MRD的分子标记，我们已经开发了灵敏的PCR分析方法 对于FLT3ITD，可用作MRD的检测。《特定目标2》审查 检测Flt3ITDS的MRD对预测患者临床复发的能力 SWOG S0106。所有的Flt3 ITDS的MRD分析将在所有的Flt3进行 治疗期间特定时间点的阳性患者及其治疗结果 将与临床结果相关。我们假设MRD化验 使用Flt3ITDS可以预测临床复发。 具体目标三：检查参与激活的分子效应器 突变的Flt3受体，并开发技术来抵消 突变的Flt受体。转染Flt3ITD的小鼠细胞的数据 提示FLT3ITDS导致FL T3受体的激活，该受体可以 被非特异性酪氨酸激酶抑制剂抑制。我们会变身为 通过基因工程使FL T3突变进入小鼠细胞。的影响 这些Flt3突变对受体、MAP激酶、STAT3和5、c- Jun，并将研究全球遗传表达。我们还将确定 如果针对FL T3受体的抗体，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 Flt3突变的影响。急性髓系白血病患者样本的平行试验 对于Flt3，还将进行ITDS。我们假设Flt3ITDS将 受体的早期结构性激活，可被 酪氨酸激酶抑制剂。 除了这项研究提案外，斯特雷沃特博士还开发了一种 教育计划，包括具体的培训目标，说教 课程，以及他的顾问的正式审查。这项教育计划将 加强他在临床试验设计和方法学方面的背景。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调查员的技能。

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.