Understanding metabolic vulnerabilities in cancer and the impact the tumor microenvironment has on cancer progression.

了解癌症的代谢脆弱性以及肿瘤微环境对癌症进展的影响。

• 批准号: 10413112
• 负责人: Shonagh Russell
• 金额: $ 9.32万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-22 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10413112
• 关键词: 3-Dimensional; ATP phosphohydrolase; Acids; Active Sites; Affect; Area; Automobile Driving; Bicarbonates; Biological Assay; Biological Models; Breast; Breast Cancer Cell; Breast Cancer cell line; Buffers; Cancer Patient; Carbon Dioxide; Cell Line; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Consumption; Data; Disease; Disseminated Malignant Neoplasm; Doctor of Philosophy; Dyes; Ecology; Evolution; Fellowship; Fermentation; Fluorescence; Genotype; Genus Hippocampus; Glucose; Glycolysis; Grant; Human; Hydration status; Hypoxia; Image; Injections; Institution; Label; Lactic acid; MCF7 cell; Malignant Neoplasms; Manuscripts; Measurement; Measures; Membrane; Membrane Potentials; Membrane Transport Proteins; Metabolic; Metabolic Pathway; Metabolism; Metastatic Neoplasm to the Lung; Methodology; Mitochondria; Modeling; Nature; Needle biopsy procedure; Neoplasm Metastasis; Noninfiltrating Intraductal Carcinoma; Nonmetastatic; Outcome; Ovarian; Oxides; Oxygen; Oxygen Consumption; Patient-Focused Outcomes; Patients; Persons; Phenocopy; Phenotype; Postdoctoral Fellow; Production; Protein Export Pathway; Proteins; Protons; Reporting; Research; Research Personnel; Resistance; Role; SCID Beige Mouse; Sampling; Sodium Bicarbonate; Stress Tests; T47D; Tail; Testing; Training; Translational Research; Universities; Up-Regulation; Veins; Warburg Effect; Work; Yeasts; aerobic glycolysis; anticancer research; cancer cell; cancer type; carbonate dehydratase; career; clinically relevant; enzyme activity; experimental study; extracellular; glucose uptake; improved; improved outcome; in vivo; inhibitor; insight; lung small cell carcinoma; metabolic abnormality assessment; metabolic phenotype; metabolomics; mouse model; multidisciplinary; neoplastic cell; normoxia; overexpression; pre-doctoral; prognostic; response; skills; targeted treatment; therapeutic target; therapy resistant; three dimensional cell culture; tool; treatment response; tumor; tumor metabolism; tumor microenvironment; tumor progression; tumorigenic

Project Summary This proposal seeks to understand the impact of metabolism on cancer aggressiveness, and how metabolic vulnerabilities can be targeted to improve patient outcome. Although targeted therapies have a great focus in the cancer research community, they have failed to generate durable responses, because of the emergence of resistance and the evolution of cancer. Metabolism is exquisitely sensitive to perturbations in the microenvironment, and this is currently an under-investigated area in cancer research. Successfully targeting metabolism has the potential to benefit patients across multiple cancer types, and genotypes, which until now has been a challenge. Cancer preferentially consumes glucose even in the presence of adequate oxygen (aerobic glycolysis), which results in the lactic acid production that decreases extracellular pH. In this Ph.D. project, it is hypothesized an alternative explanation for aerobic glycolysis, also known as the Warburg Effect (W.E), is that the enhanced uptake of glucose is due to the expression of acid exporting membrane transporters. Carbonic anhydrase IX (CA-IX) is one such acid producing protein, which we hypothesize leads to an intracellular proton deficit, driving the fermentation of glucose to replenish the deficit. CA-IX is a clinically relevant protein upregulated in numerous cancers, including breast and ovarian. CA-IX has an exofacial active site that reversibly hydrates CO2 into HCO3- and H+, and we term it a pseudohypoxic protein, as although regulated by hypoxia it is often expressed under normoxic conditions. In Aim 1.1 (prior studies), we have shown that CA-IX, or PMA1(yeast proton ATPase), over-expression in a lowly aggressive, non-metastatic breast cancer cell line (MCF-7) increases the glycolytic rate, glucose uptake, lactate production and increases lung metastasis in vivo. We also developed a metabolic profiling tool to compare 2D and 3D metabolism in the Seahorse Extracellular Flux Analyzer to aid us in our metabolic studies. Finally, preliminary pHi studies show our CA-IX clones have a higher intracellular pH compared to parental MCF-7. In Aim 1.2, (proposed studies), we will take more robust measurements of intracellular pH in both CA-IX and PMA-1 clones at various extracellular pH. We will also measure CA-IX enzymatic activity in the presence and absence of a CA-IX inhibitor from Philogen. We will repeat our in vivo tail vein experimental metastasis studies in the presence and absence of sodium bicarbonate (buffer therapy), and the CA-IX Philogen inhibitor, to see if it reduces metastasis. Finally, in Aim 2, the post-doc will focus on understanding the metabolic phenotype of small cell lung cancer (SCLC). The work will be translationally focused to aid in the treatment of patients. SCLC has a very poor prognostic outcome and, currently, its metabolic vulnerabilities are untapped. Using 13C-labelled metabolite studies in vivo and through patient needle biopsies, we hope to elucidate those metabolic pathways aiding in aggressiveness of the disease. Overall, the focus of my career is to understand the impact of cancer cell metabolism on patient outcome and therapy resistance to clinically benefit patients.

项目摘要 该提议旨在了解代谢对癌症侵略性的影响以及如何代谢 可以针对改善患者预后的漏洞。尽管有针对性的疗法非常重点 癌症研究界，由于出现 抗药性和癌症的进化。代谢对扰动非常敏感 微环境，目前是癌症研究中受访的领域不足的领域。成功定位 代谢有可能使多种癌症类型和基因型的患者受益，直到现在 一直是一个挑战。即使在有足够的氧气的情况下，癌症也优先消耗葡萄糖 （有氧糖酵解），导致乳酸产生可降低细胞外pH。在此博士学位 项目，这是有氧糖酵解的另一种解释，也称为Warburg效应 （w.e），是由于酸的表达导出的葡萄糖的摄取增强 运输商。碳酸酐酶IX（CA-IX）是一种产生酸的蛋白质，我们假设铅是 在细胞内质子不足，驱动葡萄糖的发酵以补充赤字。 CA-IX在临床上是 相关的蛋白质在包括乳房和卵巢在内的许多癌症中上调。 Ca-ix具有外叶活性 可逆地将二氧化碳水合到HCO3和H+中的位置，我们将其称为假氧蛋白，仿佛 受缺氧调节通常在常氧条件下表达。在AIM 1.1（先前的研究）中，我们有 表明Ca-Ix或PMA1（酵母质子ATPase）在低侵略性的非转移性中过表达 乳腺癌细胞系（MCF-7）增加了糖酵解速率，葡萄糖摄取，乳酸产生并增加 体内肺转移。我们还开发了一种代谢分析工具来比较2D和3D代谢 海马细胞外通量分析仪有助于我们进行代谢研究。最后，初步PHI研究表明 与亲本MCF-7相比，我们的CA-IX克隆具有更高的细胞内pH值。在AIM 1.2（拟议的研究）中， 我们将在各种CA-IX和PMA-1克隆中对细胞内pH进行更强大的测量 细胞外pH。我们还将在存在和不存在CA-IX的情况下测量CA-IX酶活性 来自哲学家的抑制剂。我们将在存在的情况下重复体内静脉静脉实验转移研究 缺乏碳酸氢钠（缓冲疗法）和CA-IX Philogen抑制剂，以查看它是否减少 转移。最后，在AIM 2中，DOC将专注于理解小细胞的代谢表型 肺癌（SCLC）。这项工作将集中在翻译上，以帮助治疗患者。 SCLC有一个 预后的结果非常差，目前尚未开发其代谢脆弱性。使用13C标记 代谢物研究在体内和通过患者针头活检，我们希望阐明这些代谢途径 协助疾病的侵略性。总体而言，我职业的重点是了解癌症的影响 细胞代谢对临床上的患者抗药性和耐药性有益。