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

基本信息

批准号：
10090748
负责人：
Younghye Song
金额：
$ 22.5万
依托单位：
UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
依托单位国家：
美国
项目类别：
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/10090748
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/antagonist 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

项目摘要

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.

项目总结 – 项目负责人 Young Hye Song 该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最近的临床数据分析表明，神经纤维与脑转移的关系。与导管原位癌和正常导管癌相比，浸润性导管癌中存在更多有证据表明谷氨酰胺代谢可能促进乳腺肿瘤神经支配和大脑。转移：1）乳腺癌细胞表达高水平的谷氨酰胺酶，通过转化来启动谷氨酰胺分解谷氨酰胺转化为谷氨酸，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对谷氨酰胺的需求，3) 糖酵解和谷氨酰胺分解之间的协同作用增强癌症细胞将谷氨酰胺转化为乳酸，刺激脑源性神经营养因子（BDNF）的分泌驱动神经突延伸，4) 脑转移乳腺癌细胞表达突触后标记 PSD-95 和然而，BDNF 受体 TrkB 具有抑制这几个关键代谢途径并减少转移的潜力。三阴性乳腺癌的表型尚未明确阐明为此，我们对此进行了斗争。三阴性乳腺癌细胞提高的谷氨酰胺代谢促进乳腺肿瘤神经支配为了验证这一假设，我们将对 3D 培养平台进行生物工程设计，以紧密模仿乳腺。为此，我们将开发乳腺肿瘤的组织工程模型。以脱细胞乳腺组织为支架培养4T1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)（目的 1) 我们将使用状态分析乳腺肿瘤神经支配 3D 模型的谷氨酰胺代谢谱。最先进的代谢分析工具并利用代谢辅因子的内源性自发荧光（Aim 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 4T1 和 ASC 的体外和体内植入（目标 3）。将测试合成、消耗和谷氨酸生产/功能的阻断神经支配和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 体积延时共焦和多光子成像，以及用于代谢分析的 Bioenergetics Core 体外培养）以在研究设计、执行、分析和解释方面实现强大的灵活性并改进结合我们对乳腺肿瘤神经支配和脑转移的理解以及蒂姆博士的指导。 Muldoon 和 Robert Griffin 博士，以及 AIMRC 的高级指导委员会和其他中心领导宋博士将为科学独立建立一条坚实的道路，并在年底提交 R01 第二年侧重于全面研究，与目标 3 的重点和长期项目目标相呼应。