METABOLOMIC ANALYSES OF HER2-POSITIVE BREAST CANCER CELLS

项目总结 致癌受体HER2在25%-30%的乳腺癌患者中过度表达，其特征是 侵袭性生长，疾病复发增加，预后不良。虽然大部分的信号 机制(S)已阐明，HER2阳性对细胞代谢的影响不是很好 明白了。最近，我们发现细胞内乳酸的高积聚与HER2阳性有关。 乳腺癌细胞。乳酸在肿瘤组织内的积聚是众所周知的(Warburg效应)，主要是由于 癌细胞糖酵解率增加。乳酸脱氢酶的主要酶(LDHA和LDHB)是 负责丙酮酸向乳酸的转化，常在癌症中过度表达。这一证据表明 令人担忧的临床问题：HER2阳性是导致新陈代谢脱轨的驱动因素吗？ 增殖/侵袭性表型？我们假设HER2的表达是糖酵解的驱动因素 导致乳酸盐积聚的乳腺癌亚群的表型。我们提出了三个相互关联的目标 系统研究HER2过表达对乳腺癌细胞存活的生化影响 通过识别和跟踪乳酸糖酵解途径中的关键代谢物，了解如何对抗 HER2策略在特定的实验细胞环境中促进细胞死亡：1)决定代谢路径 HER2阳性乳腺癌细胞中乳酸的变化，2)确定抗HER2策略的代谢影响，以及 3)确定目标候选基因在改变的代谢途径中的表达。3D/球体乳房的组合 具有可变和可诱导的HER2表达的癌症模型将与代谢示踪剂(13C)-葡萄糖、- 通过无偏的、数据驱动的核磁共振来确定乳酸、谷氨酰胺随时间的消耗和释放情况 光谱学。这一提议意义重大，因为它试图揭示HER2和HER2之间的分子机制 和乳酸使用代谢组学方法在稳健的等基因3D培养中。结果可能会直接影响到 选择蛋白质和小分子药理工具更好地了解乳酸在人体内的转运 恶性肿瘤。此外，这是为了应对有限的范围，URM学生的学习目标是SC3 主动性和关键路径必须通过基于体外的球体研究进行概念验证 然后再转化为先进的模型，如人类乳腺癌样本。 。

PROJECT SUMMARY The oncogenic receptor, HER2, is overexpressed in 25-30% of breast cancer patients and is characterized by aggressive growth, increased disease recurrence, and poor prognosis. While much of the signaling mechanism(s) have been elucidated, the impact of HER2 positivity on cellular metabolism is not well understood. Recently, we found high intracellular accumulation of lactate that correlated with HER2 positivity in breast cancer cells. Lactate accumulation within tumor tissue is well known (Warburg effect) and largely due to the increased glycolytic rate of cancer cells. The major lactate dehydrogenase enzymes (LDHA and LDHB) are responsible for conversion of pyruvate to lactate and often overexpressed in cancer. This evidence points to the alarming clinical problem: is HER2 positivity a driver towards derailed metabolism and a potentially more proliferative/aggressive phenotype? We hypothesize that HER2 expression is the driver for a glycolytic phenotype in a subset of breast cancer that leads to lactate accumulation. We propose three interlinked aims to systematically define the biochemical impact of HER2 overexpression to drive breast cancer cell survival through identifying and tracking key metabolites in the glycolytic pathway of lactate, and understand how anti- HER2 strategies promote cell death in specific experimental cellular contexts: 1) determine the metabolic path of lactate in HER2-positive breast cancer cells, 2) determine the metabolic impact of anti-HER2 strategies, and 3) define target candidate expression in altered metabolic pathways. A combination of 3D/spheroid breast cancer models with variable and inducible HER2 expression will be used with metabolic tracers (13C)-glucose, - lactate, -glutamine to define consumption and release profiles over time by unbiased, data-driven NMR spectroscopy. This proposal is significant because it seeks to reveal the molecular mechanism between HER2 and lactate using a metabolomic approach in a robust isogenic 3D culture. Outcomes may directly impact the selection of protein and small molecule pharmacologic tools to better understand lactate shuttling in human malignancies. Furthermore, this is responsive to the limited scope, URM student studies targeted for the SC3 initiative and it is imperative for the critical path to show proof-of-concept with in vitro-based spheroid studies before translating into advanced models such as human breast cancer samples. .