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。

基本信息

批准号：
10644011
负责人：
Shonagh Russell
金额：
$ 9.75万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
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/10644011
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 Cell secretion 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 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 comparison control enzyme activity experimental study extracellular glucose uptake improved improved outcome in vivo inhibitor insight metabolic abnormality assessment metabolic phenotype metabolomics mouse model multidisciplinary neoplastic cell normoxia overexpression pre-doctoral prognostic response skills small cell lung carcinoma targeted treatment therapeutic target therapy resistant three dimensional cell culture tool treatment response tumor tumor metabolism tumor microenvironment tumor progression tumorigenic

项目摘要

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.

项目概要该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的治疗结果。尽管靶向治疗的重点是在癌症研究界，他们未能产生持久的反应，因为出现了耐药性和癌症的进化。新陈代谢对体内的扰动极其敏感微环境，这是目前癌症研究中一个研究不足的领域。成功瞄准新陈代谢有可能使多种癌症类型和基因型的患者受益，到目前为止，一直是一个挑战。即使有足够的氧气，癌症也会优先消耗葡萄糖（有氧糖酵解），导致乳酸产生，降低细胞外 pH 值。在这个博士学位中。项目中，假设有氧糖酵解的另一种解释，也称为瓦伯格效应 (W.E)，葡萄糖的摄取增强是由于酸输出膜的表达运输者。碳酸酐酶 IX (CA-IX) 就是这样一种产酸蛋白，我们假设它导致细胞内质子缺乏，驱动葡萄糖发酵来补充缺乏。 CA-IX 是一种临床相关蛋白质在多种癌症中上调，包括乳腺癌和卵巢癌。 CA-IX 具有面部外活性可逆地将 CO2 水合成 HCO3- 和 H+ 的位点，我们将其称为假缺氧蛋白，尽管受缺氧调节，它通常在含氧量正常的条件下表达。在目标 1.1（先前的研究）中，我们有表明 CA-IX 或 PMA1（酵母质子 ATP 酶）在低侵袭性、非转移性细胞中过度表达乳腺癌细胞系 (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来比较 2D 和 3D 代谢 Seahorse 细胞外通量分析仪帮助我们进行代谢研究。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。在目标 1.2（拟议的研究）中，我们将在不同的时间对 CA-IX 和 PMA-1 克隆的细胞内 pH 进行更可靠的测量细胞外pH值。我们还将测量存在和不存在 CA-IX 的情况下的 CA-IX 酶活性来自 Philogen 的抑制剂。我们将在存在和存在的情况下重复我们的体内尾静脉实验转移研究不使用碳酸氢钠（缓冲疗法）和 CA-IX Philogen 抑制剂，看看它是否会减少转移。最后，在目标 2 中，博士后将重点了解小细胞的代谢表型肺癌（SCLC）。这项工作将集中于帮助患者的治疗。 SCLC 有一个预后非常差，目前其代谢脆弱性尚未开发。使用 13C 标记通过体内代谢研究和患者针活检，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预后和治疗耐药性的影响使临床患者受益。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Development and Characterization of a Luciferase Labeled, Syngeneic Murine Model of Ovarian Cancer.

荧光素酶标记的同基因小鼠卵巢癌模型的开发和表征。

DOI：
发表时间：
2022-08-30
期刊：
影响因子：
5.2
作者：
Russell, Shonagh;Lim, Felicia;Peters, Pamela N;Wardell, Suzanne E;Whitaker, Regina;Chang, Ching;Previs, Rebecca A;McDonnell, Donald P
通讯作者：
McDonnell, Donald P