Identifying molecular markers that predict relapse after therapy discontinuation inchronic myeloid leukemia.

识别预测慢性粒细胞白血病治疗停止后复发的分子标记。

• 批准号: 10291794
• 负责人: Elizabeth Ann Eklund
• 金额: --
• 依托单位: JESSE BROWN VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10291794
• 关键词: Adult; Aftercare; Apoptosis; Blast Phase; Bone Marrow; Bone Marrow Cells; Bone Marrow Diseases; Bone Marrow Transplantation; Calpain; Cells; Characteristics; Chronic; Chronic Myeloid Leukemia; Chronic Phase; Clinical; Clinical Practice Guideline; Clinical Trials; Development; Disease remission; Elderly; Future; Generations; Genes; Goals; Guidelines; Healthcare Systems; Human; Imatinib; In complete remission; Incidence; Individual; Innate Immune Response; Malignant - descriptor; Modeling; Molecular; Mus; Mutation; Myeloid Leukemia; National Comprehensive Cancer Network; Newly Diagnosed; Ontology; Outcome; Pathway interactions; Patients; Population; Production; Prognostic Marker; Regulation; Relapse; Research Personnel; Resistance; Ribonucleases; Sampling; Secondary to; Signal Transduction; Transcript; Translating; Transplantation; Tyrosine Kinase Inhibitor; abl Oncogene; calpain inhibitor; cellular transduction; cost; cytokine; discontinuation study; follow-up; fusion gene; glycogen synthase kinase 3 beta inhibitor; inhibitor/antagonist; interest; leukemic stem cell; middle age; molecular marker; mouse model; patient derived xenograft model; pre-clinical; preclinical study; prevent; relapse patients; relapse prediction; relapse risk; response; side effect; single-cell RNA sequencing; standard of care; survivin; therapeutic target; tool; transcriptome; transcriptome sequencing; transplant model

Clinical outcomes in chronic myeloid leukemia (CML) were revolutionized by development of imatinib (IM) and later generation tyrosine kinase inhibitors (TKIs) that target the Bcr-abl oncogene. Currently, ~55% patients with newly diagnosed CML achieve a major molecular response to TKI treatment (MMR) and 17% a complete molecular response (CMR). Because CML patients survive many years, there is interest in identifying those who can discontinue treatment. This interest is encouraged by the 30% incidence of side effects in during long term TKI therapy and the substantial costs of chronic TKI treatment to patients and healthcare systems. Unfortunately, ~60% of subjects with sustained MMR/CMR relapsed after TKI discontinuation in several clinical trials. Given favorable rates results of TKI re-induction with after relapse, a therapy discontinuation attempt is a part of the National Comprehensive Cancer Network (NCCN) guidelines for patients with prolonged CMR. These discontinuation studies suggest some leukemia stem cells (LSCs) persist in remission. Persistent LSCs do not have mutation or duplication of the BCRABL fusion gene, as is seen with overt TKI resistance, but provide a reservoir of cells susceptible to acquiring such mutations, or those leading to blast crisis (BC). Current clinical tools are inaccurate in predicting the likelihood of relapse vs sustained remission post TKI discontinuation. We developed a murine bone marrow transplantation model to define characteristics associated with successful TKI discontinuation. In this model, primary recipients of Bcr-abl transduced syngeneic bone marrow (in chronic phase; CP) were donors to secondary recipients. Secondary recipients were treated with TKIs + other agents, and mice with an MMR were donors for tertiary recipients. Tertiary recipients were followed without treatment. We found a 64% relapse rate in recipients of bone marrow from mice with IM-induced MMR that did not correlate with number of Bcr-abl+ cells from the treated donors or Bcr-abl transcript copies/cell. In prior studies, we found increased expression of Fap1 (a Fas and Gsk3β inhibitor) contributed to Fas resistance and βcatenin/survivin activity in CML-LSCs. We found that the addition of an inhibitor of Fap1 or survivin to IM treatment prevented relapse in Bcr-abl+ bone marrow recipients. Importantly, no tertiary recipients of bone marrow from IM + Fap1 or survivin inhibitor treated mice relapsed over 24 wks of observation (equivalent to 15+ human years). We found a 50x increase Bcr-abl transcript copies/GFP+ cell in the bone marrow of mice treated with IM vs IM + Fap1 or survivin inhibitor. This reflected differences in bone marrow populations in these mice. In transcriptome analysis of bone marrow from mice treated with TKI + a survivin inhibitor, we identified differences in pathways involved in positive regulation of the innate immune response, NOD-like signaling, cytokine production, and regulation of ribonuclease activity. We hypothesize that identifying pathways which are associated with relapse will permit selection of CML subjects able to safely discontinue treatment. Such pathways may be rationale therapeutic targets to permit more subjects to discontinue treatment, or regain an MMR. We will investigate this through two Aims; Aim 1: Identify molecular markers associated with relapse in CMR-CML patients undergoing a therapy discontinuation attempt. TKI treatment will be discontinued per NCCN guidelines. A clinical trial will correlate molecular markers in the subject’s bone marrow with relapse vs sustained therapy free remission. Aim 2: Define characteristics that predispose to relapse after TKI discontinuation in a murine CML model. We will investigate molecular mechanisms predisposing to relapse after TKI discontinuation in patient derived xenografts or murine bone marrow transplants. Implicated pathways will be targeted in pre-clinical studies. In these studies, we employ a pre-clinical murine model to study TKI discontinuation. Results will be translated to inform studies in human CML subjects. The goal is to identify pathways associated with relapse after TKI discontinuation as prognostic indicators and potential future therapeutic targets.

慢性髓样白血病（CML）的临床结果通过伊马替尼（IM）的发展而彻底改变 以及后来的一代酪氨酸激酶抑制剂（TKI），靶向BCR-ABL癌基因。目前，患者约为55％ 通过新诊断的CML实现对TKI治疗（MMR）的主要分子反应，而17％ 分子反应（CMR）。由于CML患者存活了很多年，因此确定那些人很有趣 可以停止治疗。长期的30％副作用事件鼓励了这种兴趣 TKI疗法和对患者和医疗保健系统的慢性TKI治疗的大量成本。很遗憾， 在几项临床试验中，TKI中断后，有持续的MMR/CMR继电器受试者〜60％。给出 退休后TKI重新诱导TKI重新诱导的结果，停用疗法是一部分 延长CMR患者的国家综合癌症网络（NCCN）指南。 这些中断研究表明，一些白血病干细胞（LSC）持续缓解。执着的 LSC没有明显的TKI抗性，但没有BCRABL融合基因的突变或重复 提供易受此类突变或导致爆炸危机的细胞（BC）的储层。当前的 临床工具在预测释放后的缓解可能性与TKI中断后持续缓解的可能性不准确。 我们开发了一个鼠骨髓移植模型，以定义与 成功中断的TKI。在此模型中，BCR-ABL转导的合成骨髓的主要接受者 （在慢性期； CP）是次要接受者的捐助者。次要接收者用TKIS +其他 代理商和带有MMR的小鼠是第三次接受者的捐助者。遵循第三级接受者没有 治疗。我们发现来自IM诱导的MMR的小鼠的骨髓接受者的继承率为64％ 与处理过的供体或BCR-ABL转录本副本/细胞的BCR-ABL+细胞数量无关。 在先前的研究中，我们发现FAP1（FAS和GSK3β抑制剂）的表达增加了FAS CML-LSC中的抗性和βCatenin/survivin活性。我们发现添加了FAP1或 Survivin至IM治疗可阻止BCR-ABL+骨髓受体中继电器。重要的是，没有三级接收者 来自IM + FAP1或Survivin抑制剂治疗的小鼠的骨髓的继电器超过24周观察 至15岁以上的人）。我们发现在小鼠的骨髓中增加了50倍的BCR-ABL转录副本/GFP+细胞 用IM + IM + FAP1或Survivin抑制剂处理。这反映了这些骨髓种群的差异 老鼠。在用TKI + A Survivin抑制剂治疗的小鼠的骨髓的转录组分析中，我们确定了 与先天免疫响应正调控的途径的差异，点头信号传导， 细胞因子的产生和核糖核酸酶活性的调节。 我们假设识别与继电器相关的途径将允许选择CML 受试者可以安全停止治疗。这样的途径可能是理由治疗目标，以允许更多 受试者终止治疗或保持MMR。我们将通过两个目标进行调查。 AIM 1：确定与接受治疗的CMR-CML患者相关的分子标记 中断尝试。根据NCCN指南将停止使用TKI治疗。临床试验将相关 该受试者的骨髓中的分子标记物具有释放与持续治疗的免疫缓解。 AIM 2：定义在鼠CML模型中停用TKI后易于缓解的特征。 我们将调查易于缓解TKI中断后的分子机制 异种移植物或鼠骨髓移植。涉及的途径将针对临床前研究。 在这些研究中，我们采用临床前鼠模型来研究中断TKI。结果将是 翻译以告知人类CML受试者的研究。目标是识别与继电器相关的途径 在TKI停用后，作为预后指标和潜在的未来治疗靶标。