Method Development for Non-invasive Determination of Frozen Biospecimen Quality

建议：1335936 PI：Aksan从患者和健康捐赠者身上采集的约10亿份生物标本（血液、尿液、唾液、支气管灌洗液样本和组织活检）目前储存在世界各地的生物储存库和生物库中。这些生物标本储存在-80 ℃的机械冷冻箱或液氮杜瓦瓶中，以供未来研究;特别是用于分子生物标志物的发现和验证。冷冻生物标本的质量由其所含分子生物标志物的稳定性决定。如果生物样本在储存过程中降解，它们所包含的分子信息就会丢失。不幸的是，在大多数诊所和医院中，生物样本的收集和处理方案尚未标准化，因此，相当大比例的收集的生物样本甚至在冷冻和储存之前就已经变质。由于处理不当和储存条件不稳定，在储存过程中会积累更多的损坏。该项目的主要目标是开发一种光谱方法，以非侵入性和非破坏性的方式评估冷冻液体生物标本，以筛选出目前储存在生物库中的数百万低质量生物标本。为了达到我们的主要目标，将通过实验确定冷冻的关键参数，这些参数导致有害的冰-大分子相互作用屈服于结构，以及分子生物标志物的活性损失。结果将解释的数学模型的帮助下，耦合传输和冻结引起的热力学现象。然后，这些信息将用于开发一种预测算法，以非破坏性的方式使用光谱方法确定冷冻生物样品的质量。在本项目中，我们将在选定的合作生物库中进行实地测试。在本项目中，我们的目标是发展科学和技术，以快速和非破坏性地筛选出储存在全国生物库中的不合格和劣质生物标本。这将大大降低成本，更重要的是，停止在研究中使用质量差的标本。该提案中产生的科学信息还将有助于制定生物保存协议，最大限度地提高生物标本的稳定性，并开发更简单，更经济，更环保的替代现有保存方法，减少生物标本储存的碳足迹。

Proposal: 1335936PI: AksanApproximately one billion biospecimens (blood, urine, saliva, bronchial lavage fluid samples and tissue biopsies) collected from patients and healthy donors are currently being stored in biorepositories and biobanks across the world. These biospecimens are stored in -80C mechanical freezers or in liquid nitrogen dewars for future research; specifically for molecular biomarker discovery and verification. The quality of the frozen biospecimens is defined by the stability of the molecular biomarkers they contain. If the biospecimens degrade during storage, the molecular information they contain is lost. Unfortunately, in most clinics and hospitals, biospecimen collection and processing protocols have not yet been standardized and therefore a significant percentage of the collected biospecimens have deteriorated even before they were frozen and stored. More damage accumulates during storage due to improper handling and unstable storage conditions. The main goal of this project is to develop a spectroscopic method to evaluate frozen liquid biospecimens non-invasively and non-destructively to screen out the millions of low quality biospecimens currently stored in biobanks. To reach our main goal, critical parameters of freezing that result in detrimental ice-macromolecule interactions yielding to structure, and activity loss in molecular biomarkers will be determined experimentally. The results will be interpreted with the help of a mathematical model, which the couples transport and thermodynamic phenomena induced by freezing. The information will then be used to develop a predictive algorithm to determine the quality of frozen biological samples using spectroscopic methods in a non-destructive fashion. Field-testing will be conducted in selected collaborating biobanks.In this project, we aim to develop the science and technology to rapidly and nondestructively screen out the substandard and inferior biospecimens that are stored in biobanks across the nation. This will significantly cut costs and more importantly, stop the use of bad quality specimens in research. The scientific information generated in this proposal will also help develop biopreservation protocols that will maximize the stability of the biological specimens, and develop simpler, economical, and greener alternatives to existing preservation methods, decreasing the carbon footprint of biospecimen storage.