Senescence-Chips for Radiation Biodosimetry

项目概要/摘要 放射性/核恐怖事件后急性和迟发性辐射损伤的理想生物标志物是 其在组织损伤表现之前出现并可测量，通常在电离后一至几天 辐射（IR）暴露。它们还应该以非侵入性或微创的方式进行测量， 例如，使用外周血样本。大剂量IR可引起急性和迟发性脑损伤， 造血组织和实体组织。造血干细胞（HSC）和间充质干细胞（MSC）是造血干细胞的重要组成部分。 对这些组织的再生和修复至关重要。MSC分化为成骨细胞、软骨细胞和成骨细胞。 脂肪细胞，而HSC是各种类型血细胞的前体。我们发现HSC和MSCs 经历IR诱导的衰老，一个稳定的有丝分裂后的状态，IR后。相反，大多数分化的血液， 细胞在IR后经历凋亡并被迅速清除。HSC和MSC通常存在于骨中， 骨髓中，但小数量总是可以检测到和分离外周血。在这个SBIR项目中， 我们建议使用细胞衰老作为辐射生物剂量测定的生物终点。我们将开发 集成的微流体芯片，称为“衰老芯片”，用于快速和准确地检测衰老细胞， 特别是，IR诱导的衰老HSC和MSC，以及循环细胞因子/趋化因子， 衰老相关分泌表型（SASP），来自少量人外周血。的 该项目建立在我们最近发表的表征IR诱导的HSC衰老的进展基础上， MSC、SASP作为一种稳定（慢性）表型的衰老细胞，以及一种新小鼠的开发 模型（p16- 3 MR C57 BL/6小鼠），使我们能够识别，跟踪-重要的是，诱导杀死-衰老 细胞在体内和意志。我们的新型微流控芯片包含多个功能模块， 计数HSC和MSC的总群体和衰老群体，直至单个细胞，并同时 检测外周血SASP。这些芯片将使用细胞系、小鼠模型和人类模型进行验证。 临床样本。衰老芯片将实现低成本、可重复、高度特异性和敏感性的多重 测量人体外周血的辐射生物剂量，从而作为一个外地部署 放射/核医学对策平台，包括紧急情况分类和医疗反应。

Project Summary/Abstract Ideal biomarkers of acute and delayed radiation injury after a radiological/nuclear terrorist incident are those that arise and are measurable prior to manifestation of tissue injuries, typically one to a few days after ionizing radiation (IR) exposure. They should also be measurable in a non-invasive or minimally invasive way -- for example, using peripheral blood samples. High-dose IR induces acute and delayed injuries to both hematopoietic and solid tissues. Hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) are critical for regenerating and repairing these tissues. MSCs differentiate into osteoblasts, chondrocytes, and adipocytes while HSCs are precursors of various types of blood cells. We showed that HSCs and MSCs undergo IR-induced senescence, a stable post-mitotic state, after IR. In contrast, most differentiated blood cells undergo apoptosis after IR and are quickly cleared. HSCs and MSCs generally reside in the bone marrow, but small numbers can always be detected and isolated from peripheral blood. In this SBIR project, we propose to use cellular senescence as the biological end-point for radiation biodosimetry. We will develop integrated microfluidic chips, termed "Senescence-Chips", for rapid and accurate detection of senescent cells, particularly, IR-induced senescent HSCs and MSCs, as well as circulating cytokines/chemokines due to the senescence-associated secretory phenotype (SASP), from small volumes of human peripheral blood. The project builds on our recently published progress in characterizing the IR-induced senescence of HSCs and MSCs, the SASP as a stable (chronic) phenotype of senescent cells, and the development of a new mouse model (p16-3MR C57BL/6 mice) that allows us to identify, track - and, importantly, inducibly kill - senescent cells in vivo and at will. Our novel microfluidic chips contain multiple functional modules that will capture and enumerate the total and senescent populations of HSCs and MSCs, down to single cells, and simultaneously detect SASP, in peripheral blood. The chips will be validated using cell lines, mouse models, and human clinical samples. Senescence-chips will enable low-cost, reproducible, highly specific and sensitive multiplex measurements of human peripheral blood for radiation biodosimetry, thus serving as a field-deployable platform for radiological/nuclear medical countermeasures including emergency triage and medical responses.