Excellence in Research: Stimuli-Responsive Exosomes for Quantitative Biosensing

卓越的研究：用于定量生物传感的刺激响应外泌体

• 批准号: 2302452
• 负责人: Kristen Dellinger
• 金额: $ 49.95万
• 依托单位: North Carolina Agricultural & Technical State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2302452&HistoricalAwards=false
• 关键词: Excellence; Research; Stimuli; Responsive; Exosomes

Exosomes are a category of lipid-bound, nanosized vesicles ( small sacs formed by a membrane and filled with liquid) produced by most cells in the body. Due to the nature of their packaging and release, these vesicles can contain molecules that are characteristic of their cells of origin. This means that exosomes can carry biomarkers for various disease states, including cancer and Alzheimer’s disease. However, current methods for isolating and purifying exosomes are costly and often ineffective. Developing new ways to control the release of exosome contents is essential to engineer better diagnostic tests. This project will investigate a new set of knowledge and develop tools to control the behavior of exosomes through the application of heat. Students participating in this multidisciplinary research will be mentored and trained to prepare them for a successful career in STEM through summer research experiences, conferences, and hands-on lab experiences. The goal of this Historically Black Colleges and Universities - Excellence in Research (HBCU-EiR) project is to develop and investigate the potential for engineering thermo-sensitive liposomes for applications in biosensing and downstream diagnostic applications. This will address a critical challenge in the field, which is to exert selective control over specific exosome populations within a mixed sample. More specifically, the ability to direct the release of a subset of exosomes from a capture substrate and to discharge their intra-vesicular cargo in a reliable and highly controlled manner could replace burdensome purification and isolation protocols and broaden the technological toolbox for integrated biosensors. Inspired by recent success in engineering stimuli-responsive liposomes, the investigators propose a new paradigm in the field of exosome research. Studies are designed to test the hypothesis that exosomes can be optimized to exhibit controllable behavior (e.g., release cargo) when exposed to an external stimulus (e.g., temperature). Ultimately, the project aims to define the criteria for achieving differential release in two distinct scenarios (1) from release of a 2D captured substrate and (2) from release of intra-exosomal contents (or target analytes). This will be accomplished via the following objectives: (1) Determine whether compositional differences in cell-derived model exosomes can be used to incite stimuli-responsive behavior; and (2) Engineer thermo-responsive exosomes by selective surface modification and validate their potential for biosensing applications. These outcomes are expected to advance and transform how exosomes and other extracellular vesicles are manipulated in many settings by introducing a new mechanism to control their behavior selectively. The relevance of this work additionally lies in the potential to define and test proof-of-concept criteria for other stimuli-responsive technologies, including light, enzymes, and acoustics. This will open pathways for opportunities to advance the quantification, monitoring, and detection of any targets bound in exosomes and capabilities with high relevance to the biosensing community.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

外泌体是一类脂质结合的纳米囊泡（由膜形成并充满液体的小囊），由体内大多数细胞产生。由于其包装和释放的性质，这些囊泡可以含有其来源细胞的特征分子。这意味着外泌体可以携带各种疾病状态的生物标志物，包括癌症和阿尔茨海默病。然而，目前用于分离和纯化外泌体的方法是昂贵的并且通常无效。开发控制外泌体内容物释放的新方法对于设计更好的诊断测试至关重要。该项目将研究一套新的知识，并开发工具，通过应用热量来控制外泌体的行为。参加这项多学科研究的学生将接受指导和培训，通过夏季研究经验，会议和动手实验室经验为他们在STEM的成功职业生涯做好准备。这个历史上的黑人学院和大学-卓越研究（HBCU-EiR）项目的目标是开发和研究工程热敏脂质体在生物传感和下游诊断应用中的潜力。这将解决该领域的一个关键挑战，即对混合样品中的特定外泌体群体施加选择性控制。更具体地说，以可靠和高度受控的方式引导外泌体子集从捕获底物释放并排出其囊内货物的能力可以取代繁琐的纯化和分离方案，并拓宽集成生物传感器的技术工具箱。受到最近在工程刺激响应脂质体方面取得的成功的启发，研究人员提出了外泌体研究领域的新范式。设计研究以检验外泌体可以被优化以表现出可控行为（例如，释放货物）时暴露于外部刺激（例如，温度）。最终，该项目旨在定义在两种不同情况下实现差异释放的标准（1）从2D捕获底物的释放和（2）从外泌体内内容物（或目标分析物）的释放。这将通过以下目标来实现：（1）确定细胞衍生的模型外泌体中的组成差异是否可用于激发刺激响应行为;以及（2）通过选择性表面修饰工程化热响应外泌体并验证其生物传感应用的潜力。这些结果有望通过引入一种新的机制来选择性地控制外泌体和其他细胞外囊泡的行为，从而推进和改变外泌体和其他细胞外囊泡在许多环境中的操纵方式。这项工作的相关性还在于有可能定义和测试其他刺激响应技术的概念验证标准，包括光，酶和声学。这将为推进外泌体中任何目标的定量、监测和检测以及与生物传感社区高度相关的能力开辟道路。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。