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.

摘要实时、高精度测量体内分子和单原子离子的能力 将彻底改变生物医学研究和临床实践的许多方面。例如， 为临床医生提供有关电解质失衡的即时可操作信息监测， 危险的狭窄治疗窗的药物的血浆水平。为此，我们正在开发 基于电化学适配体（EAB）的传感器，一种可证明的可推广平台技术， 测量体内原位分析物浓度。使用这种技术，我们已经证明了 实时，秒级分辨测量十几种药物，代谢物和蛋白质生物标志物， 静脉，大脑，和周围组织的活大鼠和皮下空间的人类受试者的时期 长达24小时。在此基础上，我们提出了适体选择和适体工程方法， 在提高这些受体的敏感性，以小的变化，其目标配体的浓度。我们 第一种方法是克服小分子结合适体的亲和力通常很差，因此“调谐” 以最佳匹配临床感兴趣的浓度范围。为了实现这一目标，我们正在开发 前所未有的新的选择方案，包括类比选择，一种获得初始，如果 有时是低性能的，针对困难靶标的适体，以及插入-重选，其递归地（和 显著地）增加了这些初始适体的结构复杂性，并因此增加了其性能。我们 第二AIM使用过量结合能（即，解离常数比必需的低几倍 测量范围）作为引入变构的基础， 协同性，一种使结合曲线变陡的机制。在短期内， 拟议的研究将是一套高精度，体内EAB传感器对一组临床重要， 窄临床窗口药物、代谢物和电解质。我们工作的预期长期影响 然而，它的范围要广得多，因为我们的成功将确立一些方法， 可以合理地改进生物分子受体对变化的配体浓度的反应，这一发展将 积极影响许多基于受体的生物技术。