METABOLOMIC ANALYSES OF HER2-POSITIVE BREAST CANCER CELLS

项目摘要 致癌受体HER 2在25-30%的乳腺癌患者中过表达，其特征在于： 侵袭性生长、疾病复发增加和预后不良。虽然大部分的信号 尽管已经阐明了机制，但HER 2阳性对细胞代谢的影响尚不清楚 明白最近，我们发现高细胞内乳酸积累与HER 2阳性相关， 乳腺癌细胞肿瘤组织内的乳酸积累是众所周知的（瓦尔堡效应），主要是由于 癌细胞的糖酵解速率增加。主要的乳酸脱氢酶（LDHA和LDHB）是 负责将丙酮酸转化为乳酸，并且通常在癌症中过表达。这些证据表明， 令人担忧的临床问题：HER 2阳性是否是导致代谢脱轨的驱动因素， 增殖/侵袭表型？我们假设HER 2表达是糖酵解的驱动因素， 在乳腺癌的一个子集表型，导致乳酸积累。我们提出三个相互关联的目标 系统定义HER 2过表达对乳腺癌细胞存活的生物化学影响 通过识别和跟踪乳酸糖酵解途径中的关键代谢物，并了解抗- HER 2策略在特定实验细胞环境中促进细胞死亡：1）确定代谢途径 2）确定抗HER 2策略的代谢影响，和 3)在改变的代谢途径中定义目标候选表达。3D/球体乳房的组合 具有可变和诱导型HER 2表达的癌症模型将与代谢示踪剂（13 C）-葡萄糖， 通过无偏的数据驱动NMR确定随时间推移的消耗和释放曲线 谱这一提议是重要的，因为它试图揭示HER 2 和乳酸的代谢组学方法。结果可能直接影响 选择蛋白质和小分子药理学工具以更好地了解人体内乳酸穿梭 恶性肿瘤。此外，这是响应有限的范围，URM学生的研究目标是SC 3 这是一项倡议，关键路径必须通过体外球体研究证明概念 然后转化为先进的模型，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. .