Development of Analytical Tools for Concentration and Real-Time Control of Dissolved Gases and Their Regulation of Tissue Function

溶解气体浓度和实时控制及其组织功能调节分析工具的开发

• 批准号: 10567233
• 负责人: John C. Kramlich
• 金额: $ 48.6万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10567233
• 关键词: Acute; Anti-Inflammatory Agents; Astronauts; Atherosclerosis; Binding; Biochemical; Bioenergetics; Biological; Biological Sciences; Blood flow; Blood gas; Carbon Dioxide; Cells; Chemicals; Clinical; Collaborations; Complex; Culture Media; Cytoprotection; Data; Data Reporting; Dependence; Development; Devices; Diabetes Mellitus; Engineering; Equilibrium; Evaluation; Exposure to; Future; Gases; Goals; Graft Rejection; Heart Rate; Hormone secretion; Hormones; Hypothalamic structure; Immune response; Industry; Institution; Islets of Langerhans; Kinetics; Malignant Neoplasms; Measures; Mechanics; Mediating; Metabolic Pathway; Methodology; Methods; Modality; NADH; Neurotransmitters; Oils; Oxygen Consumption; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Physiological; Play; Process; Production; Property; Proteins; Protocols documentation; Provider; Publishing; Reaction Time; Regulation; Research; Research Personnel; Retina; Role; Sampling; Sepsis; Signal Transduction; Signaling Molecule; Solubility; Specific qualifier value; System; Technology; Testing; Time; Tissue Microarray; Tissue Model; Tissues; Toxic Environmental Substances; Tube; Validation; Water; Work; analytical tool; aqueous; design; design and construction; experience; fundamental research; instrument; instrumentation; insulin secretion; mathematical model; novel; protein purification; response; technology development; temporal measurement; therapeutic evaluation; therapy development; tool; user-friendly

Project Summary The major goal of the proposed research is to develop tools to enable the study of dissolved gases on biological samples. Both blood gases (O2, and CO2) as well as trace signaling gases (NO, CO and H2S) play critical roles in regulation of wide array of tissue functions as diverse as regulation of heart rate, blood flow, immune responses, hormone and neurotransmitter secretion, as well as cytoprotective and anti-inflammatory properties. Indeed, most if not all tissues produce and are regulated by NO, CO and H2S on top of the central role of O2 in regulating bioenergetics and metabolic pathways. Research by academic institutions as well as pharmaceutical companies are endeavoring to harness the beneficial effects of gas signals in order to treat a range of conditions including diabetes, transplant rejection, sepsis, atherosclerosis and cancer. Despite the scientific and clinical importance of dissolved gases, quantitative methods to measure real time effects of dissolved gases on tissue/cells are not available. Investigators who have studied the beneficial signaling initiated by trace gases and/or who are developing drugs to trigger the same benefits almost exclusively use water soluble surrogates/donors of each signaling gas. A critical point for this proposal is that such chemical donors of gases may not allow for accurate control of gas levels within tissue, and due to their low aqueous solubility, rapidly deplete from culture media. Preliminary data we have generated and published revealed opposite effects of dissolved H2S vs. that obtained with an aqueous provider of H2S, indicating a need to re- evaluation the effects of what is established regarding NO, H2S and CO from studies using donor molecules. As most life science researchers do not have the ability to study the direct effects of trace gases at physiologically relevant concentrations, we will develop a turnkey, automated instrumentation to control the concentration and exposure time of tissue to media containing user-specified levels of 6 gases including NO, CO, H2S, O2, CO2 and N2. This automated and user-friendly system will be constructed to supply gas mixtures to a variety of widely used tissue assessment modalities including fluidics systems, static culture in plates, and cuvette systems for studying gas binding interactions to purified proteins. We have assembled a team consisting of Bio- and Mechanical/Combustion Engineers with many years of experience designing and constructing gas fluidics system applied to biological analysis, as well as an Applied Mathematician, an Analytical Chemists and Cellular Physiologists who will be involved with the validation of the instrumentation. This technology will have broad impact on fundamental research by facilitating the study of kinetic and concentration-dependent effects of signaling gases on tissue function, signaling and bioenergetics, and evaluation of therapeutics being developed to mimic the cytoprotective properties of the gases.

项目摘要 拟议研究的主要目标是开发工具，以使溶解气体的研究能够在 生物样品。血液气体（O2和CO2）以及痕量信号传导气（NO，CO和H2S）均发挥作用 在调节广泛组织的调节中的关键作用与心率，血流调节一样多样化， 免疫反应，激素和神经递质分泌以及细胞保护和抗炎 特性。实际上，大多数组织（如果不是全部）会产生并由中央顶部的NO，CO和H2进行调节 O2在调节生物能学和代谢途径中的作用。学术机构以及 制药公司正在努力利用气体信号的有益影响，以便治疗 包括糖尿病，移植排斥，败血症，动脉粥样硬化和癌症在内的条件范围。尽管有 溶解气体的科学和临床重要性，测量实时影响的定量方法 没有组织/细胞上的溶解气体。研究了有益信号的调查人员 通过微量气和/或正在开发药物以触发相同好处的人发起的几乎专门使用 每个信号气体的水溶性替代/供体。该提议的关键点是这种化学 气体的供体可能无法准确控制组织内的气体水平，并且由于其低水位 溶解度，迅速从培养基中耗尽。我们生成和发布的初步数据揭示了 溶解的H2S与用H2S水提供者获得的相反的影响，表明需要重新 评估使用供体分子的研究中关于NO，H2S和CO的建立的影响。 由于大多数生命科学，研究人员没有能力研究痕量气体的直接影响 生理上相关的浓度，我们将开发一种交钥匙，自动化的仪器以控制 组织的浓度和暴露时间在含有用户指定水平的6个气体的培养基中 包括NO，CO，H2S，O2，CO2和N2。这个自动化和用户友好的系统将被构建为 向各种广泛使用的组织评估方式提供气体混合物，包括流体系统，静态 板中的培养物和用于研究气体结合相互作用与纯化蛋白质的培养物。我们有 组建了一个由生物和机械/燃烧工程师组成的团队，并拥有多年的经验 设计和构造气体系统应用于生物分析，以及应用的 数学家，分析性化学家和细胞生理学家，将参与验证 仪器。这项技术将通过促进研究对基础研究产生广泛的影响 信号气体对组织功能，信号传导和生物能学的动力学和浓度依赖性影响， 并评估用于模仿气体的细胞保护特性的治疗剂。