METHODS: We employed mantle cell lymphoma (MCL) cell lines demonstrating markedly diverse sensitivity of inhibition of Bruton\'s tyrosine kinase (BTK) regarding their growth and studied in-depth effects of the inhibition on various aspects of cell metabolism including metabolite synthesis using metabolomics, glucose and oxidative metabolism by Seahorse XF technology, and concentration of index metabolites lactate, alanine, total choline and taurine by 1H MRS.
RESULTS: Effective BTK inhibition profoundly suppressed key cell metabolic pathways, foremost pyrimidine and purine synthesis, the citrate (TCA) cycle, glycolysis, and pyruvate and glutamine/alanine metabolism. It also inhibited glycolysis and amino acid-related oxidative metabolism. Finally, it profoundly and quickly decreased concentration of lactate (a product of mainly glycolysis) and alanine (an indicator of amino acid metabolism) and, less universally total choline both in vitro and in vivo, in the MCL xenotransplant model. The decrease correlated directly with the degree of inhibition of lymphoma cell expansion and tumor growth.
CONCLUSIONS: Our results indicate that BTK inhibition exerts a broad and profound suppressive effect on cell metabolism and that the affected index metabolites such as lactate, alanine may serve as early, sensitive, and reliable biomarkers of inhibition in lymphoma patients detectable by noninvasive MRS-based imaging method. This kind of imaging-based detection may also be applicable to other kinase inhibitors, as well as diverse lymphoid and non-lymphoid malignancies.
方法:我们采用了套细胞淋巴瘤(MCL)细胞系,这些细胞系在抑制布鲁顿酪氨酸激酶(BTK)的生长方面表现出明显不同的敏感性,并使用代谢组学研究了这种抑制作用对细胞代谢各个方面的深入影响,包括代谢物合成,通过海马XF技术进行葡萄糖和氧化代谢,和指数代谢物乳酸的浓度,丙氨酸,总胆碱和牛磺酸的1HMRS。
结果:有效的BTK抑制作用极大地抑制了关键的细胞代谢途径,首先是嘧啶和嘌呤的合成,柠檬酸盐(TCA)循环,糖酵解,和丙酮酸和谷氨酰胺/丙氨酸代谢。它还抑制糖酵解和氨基酸相关的氧化代谢。最后,它深刻而迅速地降低了乳酸(主要是糖酵解的产物)和丙氨酸(氨基酸代谢的指标)的浓度,在体外和体内,总胆碱的普遍性较低,在MCL异种移植模型中。减少与淋巴瘤细胞扩增和肿瘤生长的抑制程度直接相关。
结论:我们的结果表明,BTK抑制对细胞代谢具有广泛而深远的抑制作用,并且受影响的指标代谢产物如乳酸,丙氨酸可以作为早期,敏感,通过基于MRS的无创成像方法检测淋巴瘤患者的抑制和可靠的生物标志物。这种基于成像的检测也可能适用于其他激酶抑制剂,以及各种淋巴和非淋巴恶性肿瘤。