Engineering approaches for point-of-care diagnosis of inner ear disorders based on the detection of blood biomarkers

内耳细胞的永久性损伤会导致听力和平衡障碍。没有诊断测试可用于确定内耳中细胞损伤的确切位置。因此，对基于检测内耳循环血液生物标志物的新诊断方法的需求尚未得到满足。在拟议的研究中，我们的目标是首次开发即时护理（POC）生物传感器平台，用于快速检测和定量内耳血液生物标志物，包括特定的细胞，基因，抗体或蛋白质。我们将在开发生物标志物识别策略（如用于特定靶捕获的基于抗体、肽或适体的缀合机制）的过程中，探索POC诊断平台用于所识别的内耳生物标志物的未表征属性，设计用于血液中靶生物标志物的快速超灵敏电化学检测的单步测定，（3）便携式一次性纸装置原型，用于在小样品体积中以低浓度检测多个目标血液生物标志物。该提案的长期目标是为内耳及其他领域建立一个全面的诊断平台。最近的报道表明，很少有内耳蛋白生物标志物，如血清普雷斯廷和耳蛋白-1作为内耳疾病的指标。然而，用于准确测量这些蛋白质的检测方法是基于传统的多步耗时的定量过程。我们将设计电化学生物传感器，以实现高水平的灵敏度，信号选择性和快速性，易于校准。我们将实施与真实的样品相容的表面固定化测定，以通过DNA杂交、抗体缀合或蛋白质-蛋白质相互作用来实现靶选择性。此外，通过设计具有大表面积的高曲率纳米结构电极，我们的目标是提高小样品体积的检测限。此外，自上而下和自下而上的方法将用于开发由玻璃或纸制成的具有可寻址通道和电极的各种微流体基底。我们将使用分子合成和共轭技术，电化学制造和印刷技术以及表征方法进行研究。最终，通过检测血液生物标志物进行更准确的诊断将有助于确定内耳损伤的部位-感觉毛细胞，听觉神经元等，这将提供诊断和监测各种内耳疾病的发生和进展以及治疗效果的能力。虽然POC生物传感平台的初始设计是用于检测内耳蛋白质生物标志物，但几种临床相关情况将受益于这种快速早期诊断的方法。

Permanent damage to the inner ear cells leads to hearing and balance disorders. There is no diagnostic test available to identify the exact location of cellular damage in the inner ear. Thus, the need for new diagnostic methods based on the detection of inner ear circulating blood biomarkers is unmet. In the proposed research, we aim to develop for the first time, point-of-care (POC) biosensor platforms for the rapid detection and quantification of inner ear blood biomarkers including specific cells, genes, antibodies, or proteins. We will explore the uncharacterized attributes of the POC diagnostic platform for the identified inner ear biomarkers throughout the development of biomarker recognition strategies such as antibody, peptide, or aptamer-based conjugation mechanisms for specific target capturing, design of single-step assays for rapid ultra-sensitive electrochemical detection of the target biomarkers in blood, (3) portable disposable paper device prototype for detection of multiple target blood biomarkers at low concentration in small sample-volumes. The long-term objective of this proposal is to build a comprehensive diagnostic platform for the inner ear and beyond. Recent reports indicated few inner ear protein biomarkers, such as serum prestin and otolin-1 as indicators of inner ear disorders. However, the detection methods for accurately measuring these proteins are based on the conventional multi-step time-consuming quantification processes. We will design electrochemical biosensors to achieve high levels of sensitivity, signal selectivity, and rapidness with the ease of calibration. We will implement the surface-immobilized assays that are compatible with real samples to achieve the target selectivity through DNA hybridization, antibody conjugation, or protein-protein interactions. In addition, with the design of high-curvature nanostructured electrodes with large surface areas, we aim to improve the detection limits in small sample volumes. Furthermore, top-down and bottom-up approaches will be used to develop various microfluidic substrates made of glass or paper with addressable channels and electrodes. We will conduct our research using molecular synthesis and conjugation techniques, electrochemical fabrication and printing techniques, as well as characterization methods. Ultimately, more accurate diagnostics through the detection of blood biomarkers will help identify the sites of damage in the inner ear - sensory hair cells, auditory neurons, etc. This will provide the ability to diagnose and monitor the occurrence and progression of a variety of inner ear disorders as well as the efficacy of treatment. While the initial design of the POC biosensing platform is for the detection of inner ear protein biomarkers, several clinically relevant circumstances would benefit from such an approach for the rapid early-stage diagnosis.