Investigating the role of glutamine metabolism in breast tumor innervation and brain metastasis

研究谷氨酰胺代谢在乳腺肿瘤神经支配和脑转移中的作用

• 批准号: 10357747
• 负责人: Younghye Song
• 金额: $ 22.07万
• 依托单位: UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10357747
• 关键词: 3-Dimensional; 4T1; Adipose tissue; Arkansas; Automobile Driving; Bioenergetics; Biomedical Engineering; Brain; Brain-Derived Neurotrophic Factor; Breast Cancer Cell; Breast Cancer Model; CD44 Antigens; CD44 gene; Cell Communication; Cells; Centers of Research Excellence; Clinical Data; Coculture Techniques; Collaborations; Collection; Consumption; Data Analyses; Data Science; Data Science Core; Development; Disease; Event; Exposure to; Fatty acid glycerol esters; Fibroblasts; Glucose; Glutamate Receptor; Glutamate-Ammonia Ligase; Glutamates; Glutaminase; Glutamine; Glycolysis; Goals; Growth; Harvest; Image; Immunofluorescence Immunologic; Implant; In Vitro; Infiltration; Investigation; Lead; Link; Malignant Neoplasms; Mammary Gland Parenchyma; Mammary Neoplasms; Measures; Mediating; Mentors; Mentorship; Metabolic; Metabolic Pathway; Metabolism; Metastatic breast cancer; Metastatic malignant neoplasm to brain; Metastatic to; Methionine Sulfoximine; Mitochondria; Modeling; Molecular; Mus; Neoplasm Metastasis; Nerve Fibers; Neuraxis; Neurites; Neurotrophic Tyrosine Kinase Receptor Type 2; Noninfiltrating Intraductal Carcinoma; Optics; Oxidation-Reduction; Pathologic; Pathway interactions; Phenotype; Phosphorylation; Play; Production; Research; Research Design; Role; Signal Transduction; Solid; Spinal Ganglia; Stains; Stromal Cells; Sulfasalazine; Synapses; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Up-Regulation; Western Blotting; Work; aerobic glycolysis; cancer cell; cell type; clinically relevant; cofactor; confocal imaging; implantation; improved; in vivo; infiltrating duct carcinoma; inhibitor; lactate dehydrogenase A; malignant breast neoplasm; mammary; metabolic profile; migration; model development; multiphoton imaging; neoplastic cell; nerve supply; neurite growth; oxidation; repaired; scaffold; spectroscopic imaging; synergism; targeted treatment; three dimensional cell culture; three-dimensional modeling; tool; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor microenvironment; tumor progression

Project Summary – Project Leader Young Hye Song The overall goal of this project is to understand the role of altered glutamine metabolism in breast tumor innervation and its connection to brain metastasis. Recent analyses of clinical data have shown that nerve fibers are significantly more present in invasive ductal carcinoma compared to both ductal carcinoma in situ and normal breast tissue. There exists evidence that glutamine metabolism may promote breast tumor innervation and brain metastasis: 1) breast cancer cells express high levels of glutaminase that initiates glutaminolysis by converting glutamine to glutamate, 2) cancer-associated fibroblasts upregulate glutamine synthetase expression to meet glutamine demands of breast cancer cells, 3) a synergy between glycolysis and glutaminolysis enhances cancer cell conversion of glutamine to lactate, which stimulates brain-derived neurotrophic factor (BDNF) secretion and drive neurite extension, 4) brain metastasizing breast cancer cells express post-synaptic marker PSD-95 and BDNF receptor TrkB. Yet, the potential to inhibit these several key metabolic pathways and reduce metastatic phenotypes in triple negative breast cancer have not been clearly elucidated. To this end, we hypothesize that elevated glutamine metabolism by triple negative breast cancer cells promotes breast tumor innervation and brain metastasis. To test this hypothesis, we will bioengineer 3D culture platforms to closely mimic the mammary tumor microenvironment. To do so, we will develop a tissue engineered model of the mammary tumor microenvironment using decellularized mammary tissue as a scaffold to culture 4T1 mouse triple negative breast cancer cells, mouse primary adipose stromal cells (ASCs) and mouse primary dorsal root ganglia (DRG) (Aim 1). We will analyze glutamine metabolic profile of the 3D model of breast tumor innervation by using state-of- the-art metabolism profiling tools and exploiting the endogenous autofluorescence of metabolic cofactors (Aim 2). Finally we will assess the influence of breast tumor innervation on brain metastasis via immunofluorescence in vitro and in vivo implantation of DRG neurite-conditioned 4T1s and ASCs (Aim 3). Inhibitors of glutamine synthesis, consumption and glutamate production/function will be tested for their ability to block innervation and the results will be analyzed in the Data Science Core to evaluate glutamine effects on breast tumor innervation in vitro. We will utilize the three cores (Data Science Core for data analysis, Imaging & Spectroscopy Core for 3D volumetric time-lapse confocal and multiphoton imaging, and Bioenergetics Core for metabolic profiling of in vitro cultures) to achieve strong rigor in study design, execution, analysis and interpretation and improve our understanding of breast tumor innervation and brain metastasis. Combined with mentoring by Dr. Tim Muldoon and Dr. Robert Griffin, as well as the AIMRC’s senior mentoring committee and other center leaders Dr. Song will establish a solid pathway for scientific independence leading to an R01 submission at the end of year 2 that focuses on a full scope study echoing the focus of Aim 3 and the long-term project goals.

项目概要-项目负责人Young Hye Song 本项目的总体目标是了解谷氨酰胺代谢改变在乳腺肿瘤中的作用 神经支配及其与脑转移的关系最近的临床数据分析表明，神经纤维 与原位导管癌和正常导管癌相比， 乳房组织有证据表明，谷氨酰胺代谢可能促进乳腺肿瘤的神经支配和脑 转移：1）乳腺癌细胞表达高水平的谷氨酰胺酶，其通过转化 2）癌症相关成纤维细胞上调谷氨酰胺合成酶表达，以满足 乳腺癌细胞的谷氨酰胺需求，3）糖酵解和谷氨酰胺解之间的协同作用增强了癌症 细胞将谷氨酰胺转化为乳酸，乳酸刺激脑源性神经营养因子（BDNF）分泌， 驱动神经突延伸，4）脑转移性乳腺癌细胞表达突触后标记物PSD-95， BDNF受体TrkB。然而，抑制这几种关键代谢途径并减少转移性肿瘤的可能性仍然存在。 三阴性乳腺癌的表型尚未明确阐明。为此，我们假设， 三阴性乳腺癌细胞谷氨酰胺代谢升高促进乳腺肿瘤神经支配， 脑转移为了验证这一假设，我们将对3D培养平台进行生物工程设计，以密切模仿乳腺癌。 肿瘤微环境为此，我们将开发一种乳腺肿瘤的组织工程模型， 使用脱细胞乳腺组织作为支架培养4 T1小鼠三阴性乳腺的微环境 癌细胞、小鼠原代脂肪基质细胞（ASC）和小鼠原代背根神经节（DRG）（Aim 1）。我们将通过使用状态分析来分析乳腺肿瘤神经支配的3D模型的谷氨酰胺代谢谱， 最先进的代谢分析工具和利用代谢辅因子的内源性自发荧光（目的 2）。最后，我们将通过免疫荧光评估乳腺肿瘤神经支配对脑转移的影响 DRG神经突调节的4 T1和ASC的体外和体内植入（目的3）。谷氨酰胺抑制剂 将测试它们的合成、消耗和谷氨酸产生/功能阻断神经支配的能力， 结果将在数据科学核心中进行分析，以评估谷氨酰胺对乳腺肿瘤神经支配的影响 体外我们将利用三个核心（用于数据分析的数据科学核心，成像和光谱核心 用于3D容积延时共聚焦和多光子成像，以及用于代谢分析的Bioenergetics Core 体外培养物），以实现研究设计、执行、分析和解释的严格性，并提高 我们对乳腺肿瘤神经支配和脑转移的理解。结合蒂姆博士的指导 马尔登和罗伯特格里芬博士，以及AIMRC的高级指导委员会和其他中心领导人 博士Song将为科学独立性建立一个坚实的途径，从而在年底提交R 01申请。 第二年，重点是全面研究，反映目标3的重点和长期项目目标。