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 这项提案旨在了解代谢对癌症侵袭性的影响，以及代谢如何影响癌症的侵袭性。 可以针对漏洞来改善患者结果。尽管靶向治疗非常注重 癌症研究界，他们未能产生持久的反应，因为出现了 抵抗力和癌症的演变。新陈代谢对体内的扰动非常敏感， 微环境，这是目前癌症研究中研究不足的领域。成功瞄准 代谢有可能使多种癌症类型和基因型的患者受益， 一直是个挑战即使在氧气充足的情况下，癌症也优先消耗葡萄糖 （有氧糖酵解），这导致乳酸的产生，降低细胞外pH值。 在一个项目中，假设有氧糖酵解的另一种解释，也称为瓦尔堡效应 (W.E)葡萄糖摄取的增强是由于酸输出膜的表达 运输机碳酸酐酶IX（CA-IX）就是这样一种产酸蛋白，我们假设它导致 导致细胞内质子缺乏，驱动葡萄糖发酵以补充不足。CA-IX是一种临床 相关蛋白在许多癌症中上调，包括乳腺癌和卵巢癌。CA-IX具有外表面活性剂 一个可逆地将CO2水合为HCO 3-和H+的位点，我们称之为假缺氧蛋白， 受缺氧调节，它通常在常氧条件下表达。在目标1.1（先前的研究）中，我们有 显示CA-IX或PMA 1（酵母质子ATP酶）在低侵袭性、非转移性 乳腺癌细胞系（MCF-7）增加糖酵解速率、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，用于比较2D和3D代谢， 海马细胞外通量分析仪，以帮助我们在我们的代谢研究。最后，初步的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。在目标1.2（拟议研究）中， 我们将在不同浓度下对CA-IX和PMA-1克隆的细胞内pH值进行更可靠的测量， 我们还将在存在和不存在CA-IX酶的情况下测量CA-IX酶活性。 Philogen的抑制剂。我们将重复我们的体内尾静脉实验转移研究， 不存在碳酸氢钠（缓冲疗法）和CA-IX Philogen抑制剂，以观察其是否减少 转移最后，在目标2中，博士后将专注于了解小细胞代谢表型 肺癌（SCLC）。这项工作的重点将是帮助病人的治疗。SCLC有一个 预后结果非常差，目前，其代谢脆弱性尚未开发。使用13 C标记 通过体内代谢物研究和患者针活检，我们希望阐明这些代谢途径 有助于疾病的侵袭性。总的来说，我职业生涯的重点是了解癌症的影响 细胞代谢对患者结局和治疗耐药性的影响，使患者临床受益。