Reprogramming of creatine metabolism in breast cancer metastasis

乳腺癌转移中肌酸代谢的重编程

• 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
• 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; 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

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标志，使癌细胞能够快速增殖，入侵和 转移。已经确定了调节癌症代谢的几种关键酶。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仅举几例。无处不在的线粒体 肌酸激酶1（CKMT1）正在成为癌肌酸代谢的新型关键酶。很少的研究 迄今为止，已经研究了CKMT1在癌症中的作用，以及CKMT1在乳腺癌中的特定作用 迁移，入侵和转移仍然在很大程度上未知。为了缩小这一知识差距，我们寻求 研究肌酸代谢在乳腺癌中的重编程。我们的初步数据表明CKMT1 驱动细胞肌酸（CR）和磷酸蛋白（PCR）浓度，并激活乳液中的糖酵解 癌细胞。我们在细胞系，小鼠模型和肌酸代谢物水平的患者中始终显示 在转移性乳腺癌细胞和转移性肿瘤中，伴随CKMT1表达下调 组织。 CKMT1在转移性乳腺癌细胞中的过表达减少了迁移，侵袭和 转移，同时增加增殖和原发性肿瘤生长。 ckmt1在非转移性中的沉默 乳腺癌细胞增加了迁移和侵袭，这是通过产生活性氧发生的 上调粘附和降解因子，上皮到间质转变（EMT）的物种（ROS）， 和信号通路。在AIM 1中，我们将严格研究 重编程肌酸代谢，相关分子途径和转移癌细胞 特性。在AIM 2中，我们将评估基因/酶和相关的分子途径是否负责 重编程肌酸代谢驱动原发性肿瘤的生长和乳房模型中的转移 癌症。我们的初步数据表明，临床乳腺癌的CKMT1表达显着降低 与原发性乳腺肿瘤相比。在AIM 3中，我们将进一步研究独特的单人 我们快速尸检计划的组织微阵列（TMA）如何肌酸代谢酶表达水平 肌酸代谢产物以及相关的分子途径在乳腺癌时会受到影响 患者转移。在我们的三个目标中，我们将测试我们的整体假设，即重新编程肌酸 代谢参与驱动乳腺癌转移。我们的初步发现提供了证据表明 肌酸代谢，尤其是CKMT1，作为预后指标和潜在治疗的承诺 转移性乳腺癌的靶标。我们的建议将大大提高我们对 重编程肌酸代谢在肿瘤进展和转移中。我们将开发综合的 多重基质辅助激光解吸/电离成像和免疫组织化学方法检测 乳腺癌标本中的肌酸酶和代谢物，用于病理工作流程的未来使用。