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: 9922662
• 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/9922662
• 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.

伊马替尼(IM)的开发彻底改变了慢性粒细胞白血病(CML)的临床结果 以及针对bcr-abl癌基因的下一代酪氨酸激酶抑制剂(TKI)。目前，约55%的患者 对于新诊断的CML，对TKI治疗(MMR)取得主要分子应答，17%完全缓解 分子反应(CMR)。因为慢性粒细胞白血病患者存活多年，所以人们有兴趣确定那些 可以停止治疗。这种兴趣是由30%的副作用在长期内的发生率鼓励的 TKI治疗以及慢性TKI治疗给患者和医疗系统带来的巨大成本。不幸的是， 在多项临床试验中，持续MMR/CMR的受试者中约60%在TKI停用后复发。vt.给出 复发后再次诱导TKI的有利比率，停止治疗尝试是 国家综合癌症网络(NCCN)针对长期CMR患者的指南。 这些停用研究表明，一些白血病干细胞(LSCs)仍在缓解中。持久化 LSCs没有BCRABL融合基因的突变或复制，这与明显的TKI耐药有关，但 提供易获得此类突变或导致爆发危机(BC)的细胞的储存库。当前 临床工具在预测TKI停药后复发和持续缓解的可能性方面是不准确的。 我们开发了一种小鼠骨髓移植模型，以确定与 成功停止使用TKI。在这个模型中，bcr-abl的主要受者移植了同基因骨髓。 慢性期(CP)为二次受者的供者。二次接受TKI+OTHER治疗 有MMR的小鼠是第三次受体的捐赠者。第三次获奖者没有被跟踪 治疗。我们发现接受IM诱导的MMR小鼠的骨髓复发率为% 与处理供者的bcr-abl+细胞数或bcr-abl转录本拷贝数无关。 在先前的研究中，我们发现FAP1(一种Fas和Gsk3β抑制剂)的表达增加与Fas有关 慢性粒细胞白血病耐药与β连接素/Survivin活性的关系我们发现，Fap1或Fap1抑制剂的添加 Survivin to IM治疗预防bcr-abl+骨髓移植受者复发。重要的是，没有第三个接受者 IM+Fap1或Survivin抑制剂治疗的小鼠骨髓复发超过24周(相当于 到15年以上的人年)。我们在小鼠骨髓中发现bcr-abl转录本拷贝数/GFP+细胞增加了50倍 用IM与IM+Fap1或Survivin抑制剂治疗。这反映了这些人的骨髓群体的差异。 老鼠。在对TKI+Survivin抑制剂处理的小鼠骨髓的转录组分析中，我们发现 参与先天性免疫反应正向调节的不同途径，如点状信号， 细胞因子的产生和核糖核酸酶活性的调节。 我们假设，识别与复发相关的途径将允许选择CML 能够安全停止治疗的受试者。这样的通路可能是允许更多 受试者停止治疗，或重新获得MMR。我们将通过两个目标来调查这一点； 目的1：确定与接受治疗的CMR-CML患者复发相关的分子标志物 中止未遂。根据NCCN指南，TKI治疗将停止。一项临床试验将与 复发与持续治疗无缓解患者骨髓中的分子标志物。 目的2：在小鼠慢性粒细胞白血病模型中，确定TKI停药后易复发的特征。 我们将研究TKI患者停止TKI后复发的分子机制 异种移植或小鼠骨髓移植。牵连的通路将成为临床前研究的目标。 在这些研究中，我们使用临床前的小鼠模型来研究TKI的停用。结果将是 翻译为对人类CML受试者的研究提供信息。目标是找出与复发相关的途径 TKI后停药作为预后指标和潜在的未来治疗靶点。