Cellular responses to pH and oxygen microenvironments in a new 3D tumor model

新 3D 肿瘤模型中细胞对 pH 和氧气微环境的反应

• 批准号: 9071293
• 负责人: Jacqueline A. De Lora
• 金额: $ 2.86万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9071293
• 关键词: Acidosis; Address; Alginates; Biochemical; Biocompatible; Biological; Biological Assay; Biological Models; Bioreactors; Blood Vessels; Blood capillaries; Buffers; Cell Hypoxia; Cell Proliferation; Cell Survival; Cells; Cellular biology; Characteristics; Chemicals; Clinical; Coculture Techniques; Complex; Consumption; Controlled Study; Coupled; Cultured Cells; Detection; Devices; Encapsulated; Energy Metabolism; Environment; Enzymes; Fellowship; Fluorescence; Fluorometry; Freezing; Gene Expression; Gene Proteins; Germ Cells; Glass; Glucose Transporter; Goals; Half-Life; Harvest; Health; Hypoxia; Hypoxia Inducible Factor; Image Analysis; In Situ; In Vitro; Indium; Investigation; Lead; Length; Ligand Binding; Location; Malignant - descriptor; Measures; Mediating; Metabolic; Metabolism; Methods; Microspheres; Modeling; Molecular Analysis; Molecular Biology; Monitor; Mono-S; Neoplastic Stromal Cell; Null Lymphocytes; Nutrient; Optics; Oxygen; Pathologic Processes; Pathway interactions; Perfusion; Phenotype; Physiological; Physiological Processes; Play; Process; Production; Proliferating; Proteins; Radial; Regulation; Role; Signal Transduction; Solid Neoplasm; System; Testing; Time; Training; Tumor Biology; Tumor Volume; Variant; Vascular Endothelial Growth Factors; angiogenesis; base; calginat; cancer cell; capillary; cell type; combinatorial; cytokine; design; extracellular; fluorescence microscope; hypoxia inducible factor 1; improved; instrument; instrumentation; metabolic rate; monomer; nanosensors; neoplastic cell; protein expression; research study; response; three dimensional cell culture; three-dimensional modeling; tool; transcription factor; tumor; tumor microenvironment; wasting

 DESCRIPTION (provided by applicant): Adaptation of cancer cells to changes in the microenvironment in an expanding tumor mass is a crucial aspect of malignant progression. To proliferate, cells depend on nutrient and waste transport from surrounding vasculature that becomes less orderly as tumor volume increases, creating chemical gradients in extracellular pH (pHe) and oxygen (O2). Hypoxia inducible factor 1α (HIF-1α), a monomer of a heterodimeric transcription factor regulated by a complex network, mediates cellular responses to physiologic and pathologic processes that allow changes in angiogenesis, cell proliferation and survival, chemosensitivity, and metabolism. The activation of the glycolytic pathway even in the presence of oxygen is a characteristic of many cancer cells, leading to an increase in metabolic consumption rates that can be directly correlated to pHe and O2. HIF-1α regulates energy metabolism by activating the gene expression of glycolytic enzymes, regulatory enzymes, and glucose transporters. HIF-1α is itself post-transcriptionally regulated through blockage of proteosomal degradation as it depends on oxygen concentration. We hypothesize that pHe also plays a role in the regulation of HIF-1α by influencing the half-life of the transcription factor potentially explaining clinical observations of increased chemosensitivity even under hypoxic conditions traditionally thought to cause chemoresistance. Currently, there are no experimental platforms that provide the tools to test such a hypothesis. The fellowship-training plan, through three specific aims, will develop a new in vitro 3D cell culture perfusion system with integrated fluorescence nanosensing capabilities for in situ detection of pHe and O2 gradients. The instrument is designed to enable spatially correlated investigation of cellular responses to the measured pHe and O2 gradients by determination of metabolic consumption rates using a simple transport model. Recovering cells and supernatants from defined regions within the device allows for biochemical and molecular analysis of HIF-1α, proteins regulated by HIF-1α, cell proliferation and survival, and VEGF as adapted by cells in different microenvironments. The three aims will ultimately yield a new integrated model system that will produce advances in: 1) quantitative, manipulable in vitro 3D models of the tumor microenvironment; 2) direct spatial correlation of pHe and O2 gradients with cellular adaptations; and 3) understanding tumor cell response to gradients of pHe and O2 as a function HIF-1α.

 描述（通过应用提供）：癌细胞在扩展的肿瘤质量中改变微环境的改变是恶性进展的关键方面。为了增殖，细胞依赖于周围脉管系统中的营养和废物转运，随着肿瘤体积的增加而变得越来越有序，从而在细胞外pH（PHE）和氧气（O2）中产生化学梯度。低氧诱导因子1α（HIF-1α）是由复杂网络调节的异二聚体转录因子的单体的单体，介导了细胞对生理和病理过程的反应，从而使血管生成，细胞增殖，生存率，化学敏感性和代谢的变化变化。即使存在氧气也是许多癌细胞的特征，糖酵解途径的激活也导致代谢消耗率增加，这可以直接与PHE和O2直接相关。 HIF-1α通过激活糖酵解酶，调节酶和糖果转运蛋白的基因表达来调节能量代谢。 HIF-1α本身在转录后通过蛋白质体降解的阻塞，因为它取决于氧气浓度。我们假设PHE还通过影响转录因子的半衰期的半衰期在调节HIF-1α中起作用，即使在传统上被认为会导致化学耐药性的缺氧条件下，也可以解释化学敏感性增加的临床观察。当前，没有一些实验平台可以提供检验这种假设的工具。通过三个特定目的，奖学金训练计划将开发出一种新的体外3D细胞培养系统，具有集成的荧光纳米传感能力，用于原位检测PHE和O2梯度。该仪器旨在通过使用简单的运输模型来确定代谢消耗率，以实现细胞对测得的PHE和O2梯度的空间相关投资。从设备内定义的区域中恢复细胞和上清液，可以对HIF-1α，由HIF-1α调节的蛋白质，细胞增殖和存活以及VEGF进行生化和分子分析，以及由不同微环境中细胞改编的VEGF。这三个目标最终将产生一个新的集成模型系统，该系统将在以下方面产生进展：1）肿瘤微环境的定量，可操作的体外3D模型； 2）PHE和O2梯度与细胞适应的直接空间相关； 3）理解肿瘤细胞对PHE和O2作为功能HIF-1α梯度的反应。