Harnessing cooperativity to achieve high-precision in vivo measurements

利用协作性实现高精度体内测量

• 批准号: 10745250
• 负责人: Tod Edward Kippin
• 金额: $ 59.01万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10745250
• 关键词: Address; Affinity; Amikacin; Ammonium; Antibiotics; Area; Binding; Biological Markers; Biomedical Research; Biotechnology; Blood Glucose; Brain; Cerebrospinal Fluid; Chemicals; Clinical; Dangerousness; Development; Diabetes Mellitus; Dissociation; Dose; Dose Limiting; Drug Delivery Systems; Drug Kinetics; Drug Monitoring; Electrolyte Balance; Electrolytes; Engineering; Ensure; Feedback; Frequencies; Funding; Generations; Goals; Health; Hormones; Human; Hyponatremia; In Situ; In Vitro; Intercellular Fluid; Ions; Ligands; Lithium; Measurement; Measures; Medical; Medicine; Metabolism; Methods; Modeling; Molecular; Monitor; Needles; Organ; Outcome; Patients; Performance; Peripheral; Pharmaceutical Preparations; Physiologic pulse; Plasma; Potassium; Proteins; Provider; Rattus; Reporting; Research; Respiration; Safety; Scheme; Shapes; Sodium; Techniques; Technology; Temperature; Therapeutic; Therapeutic Index; Time; Tissues; Toxic effect; Validation; Veins; Whole Blood; Work; analog; aptamer; clinical decision-making; clinical practice; clinically relevant; design; disease diagnosis; drug efficacy; glucose monitor; human subject; improved; in vivo; interest; invention; novel; precision drugs; protein biomarkers; rational design; receptor; receptor binding; receptor sensitivity; response; sensor; small molecule; subcutaneous; success; technology platform; temporal measurement; wearable device

Summary. The ability to measure molecules and monatomic ions in the body in real-time and with high-precision would revolutionize many aspects of both biomedical research and clinical practice. It would, for example, provide clinicians with immediately actionable information monitoring regarding electrolyte imbalances, and the plasma levels of drugs of dangerous narrow therapeutic windows. To this end, we are developing Electrochemical Aptamer-Based (EAB) sensors, a demonstrably generalizable platform technology for measuring analyte concentrations in situ in the body. Using this technique, we have already demonstrated the real-time, seconds-resolved measurement of more than a dozen drugs, metabolites and protein biomarkers in the veins, brains, and peripheral tissues of live rats and the subcutaneous space of human subjects for periods of up to 24 h. Building on this, we propose here aptamer selection and aptamer-engineering approaches aimed at improving the sensitivity of these receptors to small changes in the concentration of their target ligands. Our first approach to this end is overcome the often-poor affinity of small-molecule-binding aptamers, thus “tuning” of their affinities to optimally match the concentration range of clinical interest. To achieve this, we are developing unprecedented new selection schemes, including analog-selection, an approach for obtaining initial, if sometimes low-performance, aptamers against difficult targets, and insertion-reselection, which recursively (and dramatically) increases the structural complexity, and thus the performance, of these initial aptamers. Our second aim uses the excess binding energy (i.e., dissociation constants several-fold below the necessary measurement range) afforded by these advanced selection schemes as a basis for introducing allosteric cooperativity, a mechanism that greatly steepens binding curves. In the near term, the expected outcome of the proposed research will be a suite of high-precision, in-vivo EAB sensors against a set of clinically important, narrow-clinical-window drugs, metabolites, and electrolytes. The expected long-term impact of our work however, is much broader, as our success will establish approaches by which the responsiveness of biomolecular receptors to changing ligand concentrations can be rationally improved, a development that will positively impact many receptor-based biotechnologies.

概括。实时和高精度测量体内分子和单子离子的能力 将彻底改变生物医学研究和临床实践的许多方面。例如，它将 为临床医生提供有关电解质失衡的可起作用信息，以及 危险狭窄治疗窗口的血浆药物水平。为此，我们正在发展 电化学基于基于置换的传感器，这是一种明显的可推广平台技术 测量体内的分析物浓度。使用此技术，我们已经证明了 实时，秒确定的测量多十多种药物，代谢物和蛋白质生物标志物 活大鼠的静脉，大脑和外围组织以及人类受试者的皮下空间 最多24小时。在此基础上，我们在这里提出了针对的纠正选择和纠正工程方法 在提高这些受体对靶配体浓度少量变化的敏感性方面。我们的 解决这一目的的第一种方法是克服小分子结合体的经常贫穷的亲和力，从而“调整” 它们的亲和力与临床兴趣的浓度范围匹配。为了实现这一目标，我们正在发展 前所未有的新选择方案，包括模拟选择，一种获取初始的方法，如果 有史 大幅度地提高了这些初始适体的结构复杂性，从而提高了性能。我们的 第二目的使用过多的结合能（即解离常数低于必要的 这些高级选择方案提供的测量范围）作为引入变构的基础 协作性，一种极大的钢杆结合曲线的机制。在短期内，预​​期的结果 拟议的研究将是一套高精度，体内EAB传感器，以抗临床重要的， 狭窄的窗口药物，代谢产物和电解质。我们工作的预期长期影响 但是，要广泛得多，因为我们的成功将建立方法，通过这种方法 可以合理地改善生物分子接收器以改变配体浓度，这种发展将 积极影响许多基于受体的生物技术。