Progression of DCIS to Invasive Breast Cancer through CCR2 Chemokine Signaling

摘要 在美国，每年约有5万例导管原位癌(DCIS)被诊断出来，使DCIS成为 女性诊断的最常见类型的非浸润性乳腺癌。高达20%的患者将经历 疾病复发伴发浸润性导管癌(IDC)。现代细胞和组织病理学 方法不能准确预测疾病的进展，导致患者得不到充分的治疗或过度- 接受过DCIS的治疗。我们的长期目标是确定导致IDC的关键因素，使发展成为可能 一种基于分子的方法来预测IDC的风险，以及一种更定制的方法来治疗DCIS。这个 趋化因子受体CCR2是一种G蛋白偶联受体，正常情况下表达于巨噬细胞和 在炎症和癌症进展过程中调节对CCL2的趋化反应。使用乳房 为了模拟动物DCIS的形成，我们对现有的范式提出了挑战 关于CCL2/CCR2信号，通过证明CCL2/CCR2信号对乳腺癌细胞促进DCIS 升级到IDC。R01更新项目描述了新陈代谢和c-met在CCL2/CCR2中的作用 介导的DCIS进展，并可能确定更好的DCIS进展的预测标记物，具有意义 关于减少患者过度治疗，并为开发CCR2作为治疗靶点提供理由 减少待遇不足的情况。代谢重编程是癌症的一个重要标志，但人们对此知之甚少 在早期乳腺癌中。初步研究表明CCL2/CCR2对DCIS进展的促进作用 与糖酵解和谷氨酰胺代谢增加有关，这有助于脂肪酸的合成。CCL2 介导的乳腺癌侵袭和代谢可能依赖于CCR2和c-fos之间的相互作用。 乳腺癌细胞中的MET受体酪氨酸激酶。我们假设CCL2/CCR2趋化因子信号转导 乳腺癌细胞通过c-met依赖机制增强葡萄糖和谷氨酰胺代谢 促进DCIS的发展。目的1确定CCL2、CCR2、c-Met与代谢酶的相关性 患者标本和磁共振对DCIS进展到浸润性癌的表达 DCIS孕育动物的成像/光谱学方法。目标2是确定基因的功能贡献 C-Met to CCL2/CCR2通过调节c-Met-to CCL2/CCR2介导乳腺癌的生长、存活、侵袭和代谢 在体外乳腺癌细胞和心智模型中相遇。CCR2/c-Met/SRC相互作用机制 乳腺癌细胞株将通过分子和生化方法进行检测。目标3是确定 CCL2/CCR2介导的代谢变化促进代谢变化的生化和分子机制 糖酵解、谷氨酰胺和脂肪酸合成调控乳腺癌生长、生存和侵袭 乳腺癌细胞在生化和细胞培养分析中的途径。蛋白质降解途径 潜在调节乳腺癌细胞HKII、GLS1和FASN表达的研究将通过 生化方法。

ABSTRACT About 50,000 cases of ductal carcinoma in situ (DCIS) are diagnosed in the US every year, making DCIS the most common type of non-invasive breast cancer diagnosed in women. Up to 20% of all patients will experience disease recurrence accompanied by invasive ductal carcinoma (IDC). Current cyto- and histopathological approaches do not accurately predict disease progression, resulting in patients being under-treated or over- treated for DCIS. Our long-term goals are to identify key factors that lead to IDC that will enable the development of a molecular based approach to predict the risk of IDC, and a more tailored approach to treat DCIS. The chemokine receptor CCR2 is a G Protein Coupled Receptor, which is normally expressed on macrophages and regulates chemotaxis in response to CCL2 during inflammation and cancer progression. Using a mammary intraductal injection model (MIND) to mimic DCIS formation in animals, we have challenged current paradigms on CCL2/CCR2 signaling, by demonstrating that CCL2/CCR2 signaling to breast cancer cells promotes DCIS progression to IDC. This R01 renewal project characterizes the role of metabolism and c-MET in CCL2/CCR2 mediated DCIS progression, and may identify better predictive markers for DCIS progression, with implications on reducing patient over-treatment, and provide justification for developing CCR2 as a therapeutic target to reduce under-treatment. Metabolic reprogramming is an important hallmark of cancer, but is poorly understood in early stage breast cancer. Preliminary studies indicate that CCL2/CCR2 enhancement of DCIS progression is associated with increased glycolysis and glutamine metabolism, which facilitate fatty acid synthesis. CCL2 mediated breast cancer invasiveness and metabolism may be dependent on interactions between CCR2 and c- MET receptor tyrosine kinases in breast cancer cells. We hypothesize that CCL2/CCR2 chemokine signaling in breast cancer cells enhances glucose and glutamine metabolism through c-MET dependent mechanisms to facilitate DCIS progression. Aim 1 is to determine the relevance of CCL2, CCR2, c-MET and metabolic enzyme expression to DCIS progression to invasive carcinoma using patient samples and magnetic resonance imaging/spectroscopy approaches in DCIS bearing animals. Aim 2 is to determine the functional contribution of c-MET to CCL2/CCR2 mediated breast cancer growth, survival, invasion and metabolism through modulating c- MET in breast cancer cells in vitro and in the MIND model. Mechanisms of CCR2/c-MET/SRC interactions in breast cancer cell lines will be examined through molecular and biochemical approaches. Aim 3 is to determine the biochemical and molecular mechanisms through which CCL2/CCR2 mediated metabolic changes promote breast cancer growth, survival and invasion through modulation of glycolytic, glutamine and fatty acid synthesis pathways in breast cancer cells in biochemical and cell culture assays. Protein degradation pathways that potentially regulate HKII, GLS1 and FASN expression in breast cancer cells will be examined through biochemical approaches.