Obesity, Metabolism and Breast Cancer Metastasis

肥胖、新陈代谢和乳腺癌转移

• 批准号: 10381300
• 负责人: Stephen D Hursting
• 金额: $ 11.66万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10381300
• 关键词: 4T1; Adipose tissue; Body Composition; Breast Cancer Risk Factor; Breast cancer metastasis; Cancer Biology; Cells; Dependence; Disease; Distant; Enzymes; Epithelial; Exposure to; Fibroblasts; Genus Hippocampus; Homeostasis; Immunofluorescence Immunologic; Immunohistochemistry; Intervention; Isotope Labeling; Lipids; Mass Fragmentography; Measures; Mediator of activation protein; Mesenchymal; Metabolic; Metabolism; Mitochondria; Monoclonal Antibodies; Mus; Neoplasm Metastasis; Nucleic Acids; Obesity; Obesity Epidemic; Organ; Phenotype; Proteins; Proteomics; Pyruvate Carboxylase; Recommendation; Research; Risk; Role; Serum; Testing; Tumor Tissue; Woman; cancer cell; extracellular vesicles; interest; mortality risk; mouse model; neoplastic cell; obesity prevention; transcriptome sequencing; treatment response; triple-negative invasive breast carcinoma; tumor metabolism; tumor microenvironment

Obesity, metabolic reprogramming, and breast cancer metastasis: Role of extracellular vesicles Abstract Obesity increases the risk of triple-negative breast cancer(TNBC) and its progression to metastasis. Considering that only ~ 27% of the subjects survive past 5 years when the disease has metastasized, it is critical to understand howobesity-associated changes in body composition and metabolic homeostasis are transducedby the cancer cells and distant organs. In obesity, the hypertrophic and metabolically reprogrammed adipose tissue secretes high amounts of extracellular vesicles (EVs). In cancer biology, EVs are gaining increasing interest due to their ability to impact the recipient cells' cellular programming by delivering functional molecules, including nucleic acids, proteins, lipids, and metabolites. The internalization of EVs' cargo contributes to the epithelial-to- mesenchymal (EMT) transformation of the recipient cell and the metastatic niche formation. To mitigate metastatic potential induced by obesity and increase therapeutic response in obese women, it is crucial to understand the effects of EVs-derived from the obese tumor microenvironment (TME) and their contribution to metabolic reprogramming and metastatic progression, locally and in distant organs. In this proposal, we hypothesize that EVs derived from obese versus normoweight mice regulate PC activity promoting metabolic reprogramming and metastasis. To test this hypothesis, we will use a mouse model of obesity and TNBC to determine the impact of EVs isolated from a mouse model of obesity and TNBC on PC activity, tumor metabolism, and activation of EMT. EVs will be isolated from serum, tumor, and adipose tissue. The PC content of EVs will be screened by targeted proteomics and RNAseq. Tumor cells, MetMwntlung and 4T1, and normal fibroblasts will be cultured and exposed to isolated EVs. The impact of EVs treatment on the expression of PC and EMT mediators will be measured by qPCR. The metabolic substrate dependency will be determined by isotope labeling analysis using GC-MS, and mitochondrial function will be determined by Seahorse metabolic flux analysis and respirometry. The epithelial/mesenchymal phenotype will be evaluated by immunohistochemistry and immunofluorescence using a panel of monoclonal antibodies against established EMT-related proteins. This research will elucidate the role of EVs in the crosstalk between obesity, cancer metabolism, and TNBC metastasis. The results from these studies will lead to the identification of mechanistic targets and intervention strategies to reduce the burden of obesity in women with TNBC.

肥胖、代谢重编程和乳腺癌转移：细胞外囊泡的作用 摘要 肥胖会增加三阴性乳腺癌（TNBC）及其转移的风险。考虑 只有约27%的受试者在疾病转移后存活了5年，因此至关重要的是 了解肥胖相关的身体组成和代谢稳态变化是如何通过 癌细胞和远处器官在肥胖症中，肥大和代谢重编程的脂肪组织 分泌大量的细胞外囊泡（EV）。在癌症生物学中，电动汽车越来越受到关注， 它们通过递送功能分子（包括）来影响受体细胞细胞编程的能力 核酸、蛋白质、脂质和代谢物。EV货物的内化有助于上皮细胞对 受体细胞的间充质（EMT）转化和转移性小生境形成。以减轻 肥胖诱导的转移潜力和增加肥胖妇女的治疗反应， 了解来自肥胖肿瘤微环境（TME）的EV的影响及其对 代谢重编程和转移进展，局部和远处器官。在本提案中，我们 假设来自肥胖小鼠与正常体重小鼠EV调节PC活性促进 代谢重编程和转移。为了验证这一假设，我们将使用肥胖小鼠模型， TNBC，以确定从肥胖和TNBC的小鼠模型分离的EV对PC活性、肿瘤生长和细胞增殖的影响。 代谢和EMT的激活。EV将从血清、肿瘤和脂肪组织中分离。的PC含量 的EV将通过靶向蛋白质组学和RNAseq进行筛选。肿瘤细胞，MetMwntlung和4 T1，以及正常细胞 将培养成纤维细胞并暴露于分离的EV。EV治疗对PC表达的影响 和EMT介质将通过qPCR测量。代谢底物依赖性将通过以下方式确定： 使用GC-MS进行同位素标记分析，并通过海马代谢测定线粒体功能 流量分析和呼吸测定。上皮/间充质表型将通过 免疫组织化学和免疫荧光使用一组单克隆抗体， EMT相关蛋白质。这项研究将阐明电动汽车在肥胖、癌症和癌症之间的相互作用中的作用。 代谢和TNBC转移。这些研究的结果将有助于确定 目标和干预策略，以减轻TNBC妇女的肥胖负担。