A metabolic decision point in the progression of lymphoid malignancies

尽管最近在治疗上取得了进展，但治疗淋巴恶性肿瘤，如非霍奇金淋巴瘤和 淋巴细胞性白血病仍然是一个临床挑战。虽然对预后至关重要，但调节机制 恶性淋巴细胞的运输、迁移和固体器官的浸润性尚不完全清楚。新陈代谢 灵活性在癌症进展中起着关键作用，使癌细胞的代谢需求与 特定的组织环境。然而，在淋巴癌中，人们对代谢的影响知之甚少。 关于细胞迁移和疾病进展的编程。我们的初步研究表明，恶性 淋巴细胞迁移和固体器官渗透与细胞代谢偏好密切相关。我们 发现T和B淋巴样癌细胞在异种移植模型中的迁移和器官浸润 通过对mROSlow和mROSHigh的分析确定线粒体活性氧物种(MROS) 各州。这一创新的策略允许我们分离具有不同迁移潜力的细胞来分解燃料 “增强的迁移潜力”-mROSHigh(EMP-mROSHigh)状态的偏好。我们确认葡萄糖是 一种通过激活MROS/HIF-1a信号来推动迁移的基本燃料。初始~(13)C-葡萄糖示踪 研究表明，迁移细胞中的葡萄糖代谢发生了重新编程。减少TCA中丙酮酸的氧化 细胞周期和乳酸生成的增强通过HIF-1a信号促进迁移。这些结果支持 我们的假设是丙酮酸流量的分支点是控制恶性的一个关键的“代谢决策点” 淋巴细胞迁移和器官浸润。我们预测丙酮酸流量的动态变化 线粒体氧化和转化为乳酸通过以下途径控制迁移和渗透能力 依赖MROS/HIF-1a的细胞迁移程序的转录调控。以下是具体目标 将检验这一假设： 目的1.建立丙酮酸通量作为控制恶性淋巴细胞的决策点的作用 迁移和渗透。我们将检验这一假设，即丙酮酸代谢在三氯乙酸循环之间的转变 氧化还原为乳酸是恶性淋巴细胞迁移和固体的关键关卡 通过调节MROS/HIF-1a信号来实现器官的侵袭。 目的2.确定代谢决定控制迁移行为的分子机制 指向。这一目标将检验丙酮酸流量作为代谢决策点控制迁移的假设 通过转录调控依赖于MROS/HIF1a的细胞迁移程序。我们将表演 对CLL细胞进行RNAseq分析，然后进行功能分析以确定翻译代谢的基因 重新编程为迁移潜力。 目标3：剖析增强型移行癌细胞的燃料偏好和代谢重编程 通过对慢性淋巴细胞性白血病患者体内代谢示踪。我们将检验EMP-mROSHigh细胞的假设 在CLL患者体内表现出与体外相似的重编程葡萄糖和TCA循环代谢 表型。患者将被注入13C标记的营养物质以确定丙酮酸流量和TCA循环补给 在体内栖息地的“增强的迁移潜能”CLL细胞对帮助确定治疗策略至关重要 靶向患者的EMP-mROSHigh表型。 阐明恶性淋巴细胞迁移和实体器官浸润的代谢基础 提供对疾病生物学的重要见解并发现淋巴系统的新治疗策略 与退伍军人群体高度相关的恶性肿瘤，如CLL，因为它们与橙剂有关 以及在服兵役期间接触到其他除草剂。

Despite recent therapeutic advances, curing lymphoid malignancies such as non-Hodgkin lymphoma and lymphoid leukemia remains a clinical challenge. While critical for prognosis, the mechanisms regulating malignant lymphocyte trafficking, migration, and solid organ infiltration are incompletely understood. Metabolic flexibility plays a critical role during cancer progression aligning metabolic requirements of cancer cells with specific tissue environments. In lymphoid cancers, however, little is known about the impact of metabolic programming on cell migration and disease progression. Our preliminary studies demonstrate that malignant lymphocyte migration and solid organ infiltration are tightly connected to cellular metabolic preferences. We discovered that T- and B-lymphoid cancer cell migration and organ infiltration in xenograft models is determined by mitochondrial reactive oxygen species (mROS) through analysis of mROSlow and mROShigh states. This innovative strategy permits us to isolate cells with different migratory potentials to dissect fuel preferences of the “enhanced migratory potential”-mROShigh (EMP-mROShigh) state. We identified glucose as an essential fuel driving migration through activation of mROS/HIF-1a signaling. Initial 13C-glucose tracing studies showed reprogrammed glucose metabolism in migrating cells. Reduced pyruvate oxidation in the TCA cycle and enhanced lactate generation promoted migration through HIF-1a signaling. These results support our hypothesis that the branch point in pyruvate flux is a critical “metabolic decision point” controlling malignant lymphocyte migration and organ infiltration. We predict that dynamic shifts in pyruvate flux between mitochondrial oxidation and conversion into lactate control migratory and infiltrative potential through transcriptional regulation of mROS/HIF-1a-dependent cellular migration programs. The following specific aims will test this hypothesis: Aim 1. Establish the role of pyruvate flux as a decision point to control malignant lymphocyte migration and infiltration. We will test the hypothesis that a shift in pyruvate metabolism between TCA cycle oxidation and reduction to lactate represents a critical checkpoint of malignant lymphocyte migration and solid organ infiltration through modulation of mROS/HIF-1a signaling. Aim 2. Identify the molecular mechanisms for control of migratory behavior by the metabolic decision point. This aim will test the hypothesis that pyruvate flux as a metabolic decision point controls migration through transcriptional regulation of mROS/HIF1a-dependent cellular migration programs. We will perform RNAseq analyses of CLL cells followed by functional analyses to identify genes translating metabolic reprogramming into migration potential. Aim 3: Dissect fuel preferences and metabolic reprogramming of enhanced migratory cancer cells through in vivo metabolic tracing in CLL patients. We will test the hypothesis that the EMP-mROShigh cells in CLL patients in vivo show reprogrammed glucose and TCA cycle metabolism analogous to the in vitro phenotype. Patients will be infused with 13C-labeled nutrients to determine pyruvate flux and TCA cycle fueling in “enhanced migratory potential” CLL cells in their in vivo habitat critical to help define therapeutic strategies targeting the EMP-mROShigh phenotype in patients. Elucidating the metabolic underpinnings of malignant lymphocyte migration and solid organ infiltration will provide important insight into disease biology and uncover novel treatment strategies for lymphoid malignancies such as CLL which are highly relevant to the Veteran population due to their link to Agent Orange and other herbicide exposures during military service.