Obesity, Metabolism and Breast Cancer Metastasis

肥胖、代谢重编程和乳腺癌转移：细胞外小泡的作用 摘要 肥胖增加了三阴性乳腺癌(TNBC)及其进展为转移的风险。考虑 当疾病已经转移时，只有27%的受试者存活超过5年，这一点至关重要 了解肥胖相关的身体成分和代谢平衡的变化是如何通过 癌细胞和远处的器官。在肥胖症中，肥大和代谢重新编程的脂肪组织 分泌大量细胞外小泡(EVS)。在癌症生物学方面，电动汽车正受到越来越多的关注 它们通过递送功能分子影响受体细胞的细胞编程的能力，包括 核酸、蛋白质、脂类和代谢物。电动汽车货物的内在化有助于上皮到... 受体细胞间充质(EMT)转化和转移性生态位形成。为了减轻 肥胖引起的转移潜能和提高肥胖女性的治疗反应至关重要 了解源自肥胖肿瘤微环境(TME)的EVS的影响及其对 局部和远处器官的代谢重新编程和转移进展。在这项提案中，我们 肥胖小鼠和正常体重小鼠来源的EV调节PC活性促进的假说 代谢重编程和转移。为了验证这一假设，我们将使用肥胖和 确定从肥胖小鼠模型中分离出的EVS和TNBC对PC活性、肿瘤的影响 EMT的代谢和激活。EVS将从血清、肿瘤和脂肪组织中分离出来。PC内容 的电动汽车将通过靶向蛋白质组学和RNAseq进行筛选。肿瘤细胞，MetMwntlung和4T1，正常 成纤维细胞将被培养并暴露在分离的EV中。EVS治疗对PC表达的影响 用定量聚合酶链式反应(QPCR)检测EMT介体。代谢底物的依赖性将由以下因素决定 利用GC-MS进行同位素标记分析，线粒体功能将通过海马代谢来确定 流量分析和呼吸测量。上皮/间充质表型将通过以下方法进行评估 利用一组抗ESTESTED的单抗进行免疫组织化学和免疫荧光 EMT相关蛋白。这项研究将阐明EVS在肥胖和癌症之间的串扰中的作用 代谢和TNBC转移。这些研究的结果将导致对机械的识别 降低TNBC患者肥胖负担的目标和干预策略。

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.