The Role of EpCAM Glycosylation in Breast Cancer Metastasis

EpCAM 糖基化在乳腺癌转移中的作用

基本信息

批准号：
10605857
负责人：
Nicole Marie Jenkinson
金额：
$ 5.27万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10605857
关键词：
Adhesions Affect Basement membrane Biochemistry Biological Assay Biomedical Engineering Blood Vessels Breast Breast Cancer Cell Breast Cancer Model Breast Cancer Patient Breast Cancer Treatment Breast Cancer cell line Breast Cancer therapy Breast cancer metastasis CRISPR/Cas technology Cancer Biology Carcinoma Caring Cell Adhesion Cell Adhesion Molecules Cell Line Cells Cellular biology Cessation of life Colorectal Cancer Committee Members Complement Complex Correlation Studies Cycloheximide DNA Data Development Digestion Disease Distant Distant Metastasis Education Educational workshop Epithelial Cells Female Generations Glycobiology Glycopeptides Goals Growth Human Immunologic Deficiency Syndromes Immunoprecipitation Invaded Investments Knock-out Link Liver Lung MDA-MB-468 Malignant Neoplasms Malignant neoplasm of prostate Mammary Neoplasms Mannose Mass Spectrum Analysis Mentors Mentorship Metastatic Neoplasm to Lymph Nodes Metastatic breast cancer Microscopic Molecular Biology Mus Mutate N-Glycosylation Site Neoplasm Metastasis Nodule Normal tissue morphology Nude Mice Pathologist Patients Pattern Peptides Physicians Polysaccharides Post-Translational Protein Processing Pre-Clinical Model Primary Neoplasm Property Proteins Quantitative Reverse Transcriptase PCR Role Sampling Scientist Site Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Survival Rate Techniques Testing Tissues Training Trypsin Tumor Tissue United States Universities Widespread Disease Woman Work Writing Xenograft Model cancer cell career development doctoral student effective therapy experience experimental study glycosylation in vitro Assay in vivo insight interest knock-down liquid chromatography mass spectrometry lymph nodes mRNA Expression malignant breast neoplasm mammary epithelium mass spectrometric imaging meetings member migration mouse model mutant nano-liquid chromatography new therapeutic target novel overexpression programs protein expression success symposium treatment strategy triple-negative invasive breast carcinoma tumor growth tumor metabolism tumor xenograft undergraduate student

项目摘要

PROJECT SUMMARY Breast cancer is a widespread disease that will claim the lives of over 40,000 US women in 2022 alone, with the majority of these deaths resulting from stage IV disease, in which cancer has metastasized to distant regions, as metastatic disease is difficult to treat and has few effective treatment options. Breast cancer cells metastasize by invading the basement membrane, invading into blood vessels, circulating to distant tissues, leaving the blood vessel, and colonizing that tissue. The epithelial cell adhesion molecule (EpCAM) plays a role in cell adhesion, migration, and invasion, and has been linked to several epithelial cancers, including breast, prostate, and colorectal cancer. Despite its potential to facilitate several steps of the metastatic cascade and its link to epithelial cancers, the specific role of EpCAM in metastasis remains unknown. In his study of normal tissues, primary tumors, and metastatic nodules of 17 metastatic breast cancer patients, our collaborator Dr. Pedram Argani determined that EpCAM protein expression was increased in metastatic nodules, without a corresponding increase in EpCAM mRNA expression. We therefore hypothesize that post-translational modification of EpCAM, which has three known N-glycosylation sites and no known O-glycosylation sites, stabilizes and increases the abundance of EpCAM in metastatic tissues. Our recent study of these same samples found that the abundance of N-glycans was significantly increased in metastatic tissues versus primary tumors versus normal tissues and identified 25 significantly differentially abundant N-glycans of interest. Our study also found that EpCAM protein expression was significantly increased in metastatic tissues compared to primary tumors and was significantly statistically correlated with the expression of seven N-glycans. Taken together, these data suggest that EpCAM N-glycosylation may play a role in metastatic progression. The central hypothesis of this project is that increases and changes in N-glycosylation stabilize EpCAM and allow it to facilitate metastasis. I propose to test this hypothesis through the following specific aims: Aim 1: To elucidate the impact of EpCAM N-glycosylation on EpCAM stability and adhesion in human breast cancer cells; Aim 2: To assess the impact of EpCAM N- glycosylation on metastasis in mouse models of breast cancer. These aims will be achieved through a combination of cancer biology, biochemistry, and mass spectrometry approaches in human triple-negative breast cancer cell lines and breast cancer xenograft models. This work is significant because it will elucidate the role of N-glycosylation of EpCAM in metastatic breast cancer, determine the specific N-glycosylation profile of EpCAM in metastasizing breast cancer cells, and may subsequently uncover novel therapeutic targets and strategies in metastatic breast cancer.

项目摘要乳腺癌是一种广泛的疾病这些死亡中的大多数是由IV期疾病造成的，其中癌症已转移到遥远地区，由于转移性疾病很难治疗，并且几乎没有有效的治疗选择。乳腺癌细胞转移通过入侵地下膜，入侵血管，循环到遥远的组织，留下血液血管，并定居该组织。上皮细胞粘附分子（EPCAM）在细胞粘附中起作用，迁移和入侵，并与几种上皮癌有关，包括乳房，前列腺和结直肠癌。尽管它有可能促进转移性级联的几个步骤及其与上皮的链接癌症，EPCAM在转移中的具体作用仍然未知。在他对正常组织的研究中 17名转移性乳腺癌患者的肿瘤和转移性结节，我们的合作者Pedram Argani博士确定在转移结节中增加了Epcam蛋白表达，没有相应的 EPCAM mRNA表达的增加。因此，我们假设EPCAM的翻译后修饰，它具有三个已知的N-糖基化位点，没有已知的O-糖基化位点，可以稳定并增加转移组织中的Epcam丰度。我们最近对这些样品的研究发现丰度在转移组织与原发性肿瘤与正常组织的N-聚糖和确定了25个显着差异的n-glycans。我们的研究还发现Epcam蛋白与原发性肿瘤相比，转移组织中的表达显着增加，并且显着统计上与七个N-聚糖的表达相关。综上所述，这些数据表明EPCAM N-糖基化可能在转移性进展中起作用。该项目的中心假设是增加 N-糖基化的变化稳定EPCAM并使其促进转移。我建议对此进行测试通过以下特定目的假设：目标1：阐明Epcam N-糖基化对人类乳腺癌细胞的Epcam稳定性和粘附；目的2：评估epcam n-的影响乳腺癌小鼠模型中转移的糖基化。这些目标将通过人类三阴性乳房中癌症生物学，生物化学和质谱法的结合癌细胞系和乳腺癌异种移植模型。这项工作很重要，因为它将阐明 EPCAM在转移性乳腺癌中的N-糖基化，确定EPCAM的特异性N-糖基化谱在转移乳腺癌细胞中，随后可能会发现新颖的治疗靶标和策略转移性乳腺癌。