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Functional contributions of glycogen metabolism to ovarian cancer metastasis

糖原代谢对卵巢癌转移的功能贡献

基本信息

批准号：
10094205
负责人：
Ernst Lengyel
金额：
$ 44.05万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10094205
关键词：
Adhesions Biological Assay Biology Cancer Biology Cancer cell line Cell Proliferation Cells ChIP-seq Citric Acid Cycle Coculture Techniques Complement Coupled Cultured Cells DNA DNA Methylation Data Dioxygenases Enzymes Epigenetic Process Epithelial Epithelial Cells Event Fibroblasts Gene Expression Genes Glucose-6-Phosphate Glycogen Glycogen Storage Disease Glycolysis Goals Greater sac of peritoneum Growth Hematogenous Human In Vitro Investigation Label Libraries Link MAP Kinase Gene Malignant Neoplasms Malignant neoplasm of ovary Mammalian Oviducts Mediating Metabolic Neoplasm Metastasis Omentum Organ Peritoneum Phenotype Phosphorylation Phosphotyrosine Protein Glycosylation Proteins RNA Interference Regulation Role Seeds Serous Signal Pathway Signal Transduction Site alpha ketoglutarate base cancer cell cell type clinically relevant cofactor epigenetic regulation experimental study genome-wide glucose 1 phosphate glycogen metabolism glycogenolysis glycosylation glycosyltransferase histone methylation in vivo knock-down metabolomics mouse model nano neoplastic cell novel ovarian neoplasm overexpression phosphoproteomics promoter response seal three-dimensional modeling tumor tumor growth tumor microenvironment tumor progression

项目摘要

PROJECT SUMMARY/ABSTRACT The biology of high grade serous ovarian cancer (OvCa) is distinct from that of most epithelial tumors, in that hematogenous metastases are rare. Ovarian tumors remain confined to the peritoneal cavity and primarily seed to the omentum and peritoneum. There, cancer cells interact with cancer associated fibroblasts (CAFs) which promote cancer cell proliferation, invasion, and metastasis. To study bidirectional signaling between the two cell types, we co-cultured them and used SILAC coupled with quantitative, label-free phosphoproteomics to identify phospho-tyrosine signaling events in both OvCa cells and fibroblasts. We identified activation of phosphoglucomutase 1 (PGM1) in the cancer cells, pointing towards regulation of glycogen metabolism in OvCa cells by CAFs. Further, preliminary experiments showed that glycogenolysis provides energy to cancer cells and regulates protein glycosylation and histone methylation. Based on these data, the primary hypothesis underlying this application is that CAF-mediated glycogenolysis promotes metastasis through several different mechanisms, including glycolysis, protein glycosylation, and epigenetics. In Aim I, we will explore the metabolic consequences of PGM1 activation or inhibition in both OvCa cells and normal fallopian tube epithelial cells using untargeted metabolomics and metabolic flux studies. We will use PGM knockdown cells and investigate the effects of PGM1 inhibition on adhesion/invasion/proliferation using a 3D model and a syngeneic mouse model of metastasis. We will then utilize a high-throughput RNAi library with the goal of identifying the upstream signaling pathways regulating glycogenolysis in response to CAFs. In Aim II, we will systematically study glycogen-mediated glycosylation events using human protein glycosylation arrays to identify which proteins are specifically glycosylated by CAF-mediated glycogenolysis. This investigation will be complemented by studies determining how glycosyltransferase enzymes regulate CAF- mediated glycosylation. We will then study the role of glycosyltransferases on in vivo metastasis assays using primary CAF/OvCa cells and a syngeneic mouse model of OvCa metastasis. Using targeted metabolomics data and flux analysis of CAFs co-cultured with OvCa cells, we discovered that glycogenolysis induced α- ketoglutarate (KG). We therefore propose, in Aim III, to study how glycogenolysis alters epigenetic changes in cancer cells. We will characterize genome-wide changes in histone methylation (ChIP-seq) and DNA hydroxymethylation (Nano-hmC-Seal) to identify genes epigenetically regulated by glycogenolysis. The proposed experiments aim to define the link between glycogenolysis and epigenetics to identify functional regulators of OvCa metastasis. By understanding glycogen metabolism in the tumor organ, we may be able to elucidate novel metabolic mechanisms important for metastasis, which could result in the identification of a new and clinically relevant approach to the treatment of metastatic ovarian cancer.

项目总结/文摘