Metabolic Biomarkers of Response of Mantle Cell Lymphoma to Bruton Tyrosine Kinase Inhibition

套细胞淋巴瘤对Bruton酪氨酸激酶抑制反应的代谢生物标志物 项目摘要/摘要 由于激酶抑制剂在癌症治疗中的应用越来越多，因此有必要开发非侵入性耳科手术。 采用LY检测方法监测患者治疗的活跃度。我们的目标是开发方法来解决 三个关键问题--1)早期发现肿瘤反应或抵抗治疗，特别是通过非侵入性治疗 成像技术，2)定义药物反应和耐药性的详细机制，以及3)使用这一信息- 克服抗药性的信息。对细胞信号通路的非侵入性体内监测是不可行的， 对肿瘤体积变化的监测是一个较晚且往往具有误导性的反应指标。FDG PET，而 理想的肿瘤检测是不能预测反应的，可能是因为它只评估 糖酵解途径，而肿瘤能够使用其他途径来处理其生长所需的营养物质。 我们提出了一种新的替代方案-通过监测受 这是一种激酶抑制剂。我们的基本前提是，肿瘤代谢的变化是更早和更可靠的指标- 治疗反应的影响因素比肿瘤体积的变化更重要。我们关注的是Bruton‘s酪氨酸激酶(BTK) 使用FDA批准的BTK抑制剂ibrutinib(IBR)的信号通路，以及两个第二代In- 套细胞淋巴瘤(MCL)的抑制物。我们建议使用一种全面的基因组/磷酸- 蛋白质组/代谢组学方法描述对BTK抑制的反应和抵抗机制。在……里面 初步研究评价指标MCL细胞系，其中一株(MCL-RL)对IBR高度敏感 另一种(Jeko-1)具有抗药性。初步的RNA-Seq分析表明，BTK调控了 许多基因参与了关键的代谢途径，包括糖酵解、戊糖分流、TCA循环和Glu- 肌胺溶解。通过测量IBR治疗前后这些途径的代谢流量，我们将 确定这些途径中的哪一条对反应和抵抗至关重要。我们发现在JeKo-1细胞中，Glu- 谷氨酰胺溶解是IBR抵抗的关键，可以用谷氨酰胺酶抑制剂来克服。我们已经开发出 并验证了新的13C MRS和13C LC-MS方法用于监测肿瘤转移的关键途径的通量。 LISM，并建议在体外和体内的MCL模型上使用这些方法来确定MCL的机制。 药物反应和抗药性的NISM。乳酸、丙氨酸和胆碱已成为潜在的代谢产物。 ~1H MRS可监测MCL细胞对BTK抑制反应的生物标志物 自行研制的SEL-MQC脉冲序列。在目标1中，我们将进行Re-Re的基因组和代谢研究。 MCL患者来源的细胞系和原代细胞对BTK抑制的应答和抵抗。在目标2中，我们 将利用体外1H MRS和13C MRS和LC-MS来验证和定量代谢通量的扰动 AIM 1中确定的通路是BTK抑制的潜在成像生物标志物。最后，在目标3中，我们将利用 MCL小鼠移植瘤的~1H、~(13)C和~(31)P磁共振波谱和LC-MS分析 AIMS 1和2中确定了BTK抑制的标志物。

Metabolic Biomarkers of Response of Mantle Cell Lymphoma to Bruton Tyrosine Kinase Inhibition Project Summary/Abstract Due to increased application of kinase inhibitors to cancer therapy, there is a need to develop noninvasive ear- ly detection methods to monitor the activity of treatment in patients. We aim to develop methods to address three critical issues – 1) early detection of tumor response or resistance to therapy, particularly by noninvasive imaging techniques, 2) defining detailed mechanisms of drug response and resistance, and 3) using this infor- mation to overcome drug resistance. Noninvasive in vivo monitoring of cell signaling pathways is not feasible, and monitoring of changes in tumor volume is a late and often misleading response indicator. FDG PET, while ideal for tumor detection is not response predictive, likely because it evaluates only the first two steps of the glycolytic pathway, while tumors are able to use other pathways to process nutrients required for their growth. We propose a novel alternative – to detect response by monitoring the key metabolic pathways regulated by the kinase inhibitor. Our basic premise is that changes in tumor metabolism are earlier and more reliable indi- cators of therapeutic response than changes in tumor volume. We focus on the Bruton's tyrosine kinase (BTK) signaling pathway using an FDA-approved BTK inhibitor, ibrutinib (IBR), as well as two second-generation in- hibitors of mantle cell lymphoma (MCL). We propose to use a comprehensive genomic/phospho- proteomic/metabolomic approach to delineate the mechanism of response and resistance to BTK inhibition. In preliminary studies we have evaluated index MCL cell lines, one of which (MCL-RL) is highly sensitive to IBR and another (JeKo-1) is resistant. Preliminary RNA-Seq analysis indicates that BTK modulates expression of many genes involved in key metabolic pathways including glycolysis, the pentose shunt, TCA cycle and glu- taminolysis. By measuring metabolic flux through these pathways before and after treatment with IBR, we will determine which of these pathways are critical to response and resistance. We found that in JeKo-1 cells glu- taminolysis is critical to IBR resistance and can be overcome with glutaminase inhibitors. We have developed and validated novel 13C MRS and 13C LC-MS methods to monitor flux through key pathways of tumor metabo- lism and propose to use these methods on the MCL models in in vitro and in vivo studies to define the mecha- nism of response and resistance to the drug. Lactic acid, alanine and choline have emerged as potential meta- bolic biomarkers of response of MCL cells to BTK inhibition that can be monitored by 1H MRS using the Had- Sel-MQC pulse sequence developed by us. In Aim 1, we will perform genomic and metabolomic studies of re- sponse and resistance to BTK inhibition in cell lines and primary cells derived from MCL patients. In Aim 2, we will utilize in vitro 1H MRS and 13C MRS and LC-MS to verify and quantitate perturbations of metabolic fluxes in pathways identified in Aim 1 as potential imaging biomarkers of BTK inhibition. Finally, in Aim 3 we will utilize 1H, 13C & 31P MRS imaging and LC-MS analysis in mouse xenografts of MCL to validate the metabolic bi- omarkers of BTK inhibition identified in Aims 1 & 2.