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

套细胞淋巴瘤对布鲁顿酪氨酸激酶抑制反应的代谢生物标志物 项目总结/摘要 由于激酶抑制剂在癌症治疗中的应用越来越多，因此需要开发非侵入性的耳道抑制剂。 使用检测方法来监测患者的治疗活性。我们的目标是开发方法来解决 三个关键问题：1）早期检测肿瘤对治疗的反应或抗性，特别是通过非侵入性方法， 成像技术，2）定义药物反应和耐药性的详细机制，3）使用这种信息， 克服耐药性的方法。细胞信号传导途径的非侵入性体内监测是不可行的， 并且监测肿瘤体积的变化是一种迟发的并且经常是误导性的反应指标。FDG PET，而 理想的肿瘤检测不是反应预测，可能是因为它只评估前两个步骤， 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-通过监测受以下因素调节的关键代谢途径来检测反应： 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， 肿瘤体积的变化比治疗反应的变化更重要。我们重点研究了布鲁顿酪氨酸激酶（BTK） 信号通路使用FDA批准的BTK抑制剂，伊曲替尼（IBR），以及两个第二代 套细胞淋巴瘤（MCL）的抑制剂。我们建议使用一个全面的基因组/磷酸- 蛋白质组学/代谢组学方法来描述对BTK抑制的应答和抗性机制。在 初步研究我们已经评估了指数MCL细胞系，其中之一（MCL-RL）是高度敏感的IBR 另一株（JeKo-1）为抗性株。初步的RNA-Seq分析表明BTK调节了 许多基因参与关键的代谢途径，包括糖酵解，戊糖分流，TCA循环和葡萄糖， Taminolysis。通过测量IBR治疗前后通过这些途径的代谢通量，我们将 确定这些途径中哪些对反应和抗性至关重要。我们发现在JeKo-1细胞中， 氨解对于IBR抗性是关键的，并且可以用转氨酶抑制剂克服。我们已经开发 并验证了新的13 C MRS和13 C LC-MS方法，以监测通过肿瘤代谢关键途径的通量， 并建议将这些方法用于体外和体内研究的MCL模型，以确定其机制。 对药物的反应和耐药性。乳酸、丙氨酸和胆碱已成为潜在的Meta- MCL细胞对BTK抑制的响应的生物标志物，其可以通过1H MRS使用Had- 我们研制的Sel-MQC脉冲序列。在目标1中，我们将进行基因组学和代谢组学研究， 在来源于MCL患者的细胞系和原代细胞中对BTK抑制的应答和抗性。在目标2中， 将利用体外1H MRS和13 C MRS以及LC-MS来验证和定量代谢通量的扰动， 在目标1中鉴定为BTK抑制的潜在成像生物标志物的途径。最后，在目标3中，我们将利用 在MCL的小鼠异种移植物中的1H、13 C和31 P MRS成像和LC-MS分析，以验证代谢双- 目的1和2中鉴定的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.