Reprogramming of creatine metabolism in breast cancer metastasis

乳腺癌转移中肌酸代谢的重编程

基本信息

批准号：
10569104
负责人：
Kristine Glunde
金额：
$ 36.71万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-08 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10569104
关键词：
Adhesions Affect Amino Acids Automobile Driving Autopsy Biopsy Breast Cancer Cell Breast Cancer Model Breast Cancer cell line Breast cancer metastasis Cell Line Cells Choline Kinase Clinical Collection Creatine Data Distant Metastasis Enzymes Freezing Future Gene Expression Generations Genes Glucose Glutaminase Glycolysis Human Image Immunohistochemistry Invaded Knowledge Magnetic Resonance Spectroscopy Malignant Neoplasms Mammary Neoplasms Maps Measures Metabolic Metabolic Pathway Metabolism Metastatic Neoplasm to Lymph Nodes Metastatic breast cancer Molecular Names Neoplasm Metastasis Nonmetastatic Normal tissue morphology Nucleic Acids Organ Pathology Pathway interactions Patients Phosphocreatine Primary Neoplasm Proliferating Property Quantitative Reverse Transcriptase PCR Reactive Oxygen Species Research Research Personnel Resolution Role Sampling Signal Pathway Specimen Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Stains Testing Thymidylate Synthase Tissue Microarray Tissues Tumor Tissue Visualization Western Blotting cancer cell cell behavior epithelial to mesenchymal transition experimental study gain of function in vivo magnetic resonance spectroscopy lactate dehydrogenase A lipid metabolism loss of function lymph nodes mRNA Expression malignant breast neoplasm mass spectrometric imaging migration mitochondrial creatine kinase mouse model novel overexpression prognostic indicator programs protein expression therapeutic target tumor growth tumor metabolism tumor progression

项目摘要

Metabolic reprogramming is a hallmark of cancer, enabling cancer cells to rapidly proliferate, invade, and metastasize. Several key enzymes have been identified that modulate cancer metabolism. These include enzymes in glucose, amino acid, nucleic acid, and lipid metabolism, including lactate dehydrogenase A, glutaminase 1, thymidylate synthase, and choline kinase alpha, to name just a few. Ubiquitous mitochondrial creatine kinase 1 (CKMT1) is emerging as a novel key enzyme in creatine metabolism of cancer. Few studies to date have investigated the role of CKMT1 in cancer, and the specific role of CKMT1 in breast cancer migration, invasion and metastasis remains largely unknown. To close this knowledge gap, we seek to investigate reprogramming of creatine metabolism in breast cancer. Our preliminary data show that CKMT1 drives cellular creatine (Cr) and phosphocreatine (PCr) concentrations and activates glycolysis in breast cancer cells. We consistently show in cell lines, mouse models, and patients that creatine metabolite levels along with CKMT1 expression are downregulated in metastatic breast cancer cells and metastatic tumor tissues. Overexpression of CKMT1 in metastatic breast cancer cells reduces migration, invasion, and metastasis, while increasing proliferation and primary tumor growth. Silencing of CKMT1 in nonmetastatic breast cancer cells increases migration and invasion, which occurs through generation of reactive oxygen species (ROS) that upregulate adhesion and degradative factors, epithelial-to-mesenchymal transition (EMT), and signaling pathways. In Aim 1, we will rigorously investigate the cause-and-effect relationships between reprogramming of creatine metabolism, related molecular pathways, and metastasis-driving cancer cell properties. In Aim 2, we will assess if genes/enzymes and related molecular pathways responsible for reprogramming creatine metabolism drive primary tumor growths and metastasis in mouse models of breast cancer. Our preliminary data show that CKMT1 expression was significantly decreased in clinical breast cancer metastases as compared to primary breast tumors. In Aim 3, we will further investigate in unique single-patient tissue microarrays (TMAs) from our rapid autopsy program how creatine metabolic enzyme expression levels and creatine metabolites, as well as related molecular pathways, are affected when breast cancers metastasize in patients. In our three Aims, we will test our overall hypothesis that reprogramming of creatine metabolism participates in driving breast cancer metastasis. Our preliminary findings provide evidence that creatine metabolism, and in particular CKMT1, holds promise as prognostic indicator and potential therapeutic target for metastatic breast cancer. Our proposal will significantly advance our understanding of reprogramming of creatine metabolism in tumor progression and metastasis. We will develop integrated multiplex matrix-assisted laser desorption/ionization imaging and immunohistochemistry approaches to detect creatine enzymes and metabolites in breast cancer specimens for future use in pathology workflows.

代谢重编程是癌症的一个标志，它使癌细胞能够快速增殖、侵袭和扩散。转移。已鉴定出几种调节癌症代谢的关键酶。这些包括葡萄糖、氨基酸、核酸和脂质代谢中的酶，包括乳酸脱氢酶 A，谷氨酰胺酶 1、胸苷酸合酶和胆碱激酶 α 等。无处不在的线粒体肌酸激酶 1 (CKMT1) 正在成为癌症肌酸代谢中的一种新型关键酶。很少有研究迄今为止已经研究了 CKMT1 在癌症中的作用，以及 CKMT1 在乳腺癌中的具体作用迁移、侵袭和转移仍然很大程度上未知。为了缩小这一知识差距，我们寻求研究乳腺癌中肌酸代谢的重编程。我们的初步数据显示 CKMT1 驱动细胞肌酸 (Cr) 和磷酸肌酸 (PCr) 浓度并激活乳腺糖酵解癌细胞。我们在细胞系、小鼠模型和患者中一致证明肌酸代谢水平随着 CKMT1 表达在转移性乳腺癌细胞和转移性肿瘤中下调组织。 CKMT1 在转移性乳腺癌细胞中的过度表达可减少迁移、侵袭和转移转移，同时增加增殖和原发肿瘤的生长。非转移性细胞中 CKMT1 的沉默乳腺癌细胞通过产生活性氧来增加迁移和侵袭上调粘附和降解因子、上皮间质转化 (EMT) 的物种 (ROS)、和信号通路。在目标 1 中，我们将严格调查之间的因果关系肌酸代谢、相关分子途径和驱动癌细胞转移的重编程特性。在目标 2 中，我们将评估基因/酶和相关分子途径是否负责重编程肌酸代谢驱动小鼠乳腺模型中原发性肿瘤的生长和转移癌症。我们的初步数据显示临床乳腺癌中CKMT1表达显着降低与原发性乳腺肿瘤相比，转移灶。在目标 3 中，我们将进一步研究独特的单一患者我们快速尸检程序的组织微阵列 (TMA) 肌酸代谢酶表达水平如何当乳腺癌发生时，肌酸代谢物以及相关的分子途径会受到影响患者体内发生转移。在我们的三个目标中，我们将检验我们的总体假设，即肌酸的重新编程新陈代谢参与驱动乳腺癌转移。我们的初步调查结果证明肌酸代谢，特别是 CKMT1，有望作为预后指标和潜在的治疗方法转移性乳腺癌的靶点。我们的建议将极大地增进我们的理解肿瘤进展和转移中肌酸代谢的重新编程。我们将开发一体化多重基质辅助激光解吸/电离成像和免疫组织化学方法进行检测乳腺癌样本中的肌酸酶和代谢物，以供将来在病理学工作流程中使用。