Deciphering the role of chemical signals in inflammation with open microfluidic functional assays

通过开放微流控功能分析解读化学信号在炎症中的作用

• 批准号: 10556928
• 负责人: Ashleigh Brooks Theberge
• 金额: $ 21.77万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10556928
• 关键词: Acute; Address; Affect; Autoimmune; Award; Back; Biological Assay; Biological Markers; Biological Process; Biology; Blood; Blood capillaries; Blood specimen; COVID-19; COVID-19 complications; Cell Line; Cells; Cellular biology; Chemicals; Child; Chronic; Clinic; Clinical Research; Clinics and Hospitals; Collection; Communication; Communities; Complement; Custom; Data; Data Set; Development; Devices; Diagnostic; Diagnostics Research; Disease; Disease Pathway; Disease Progression; Economic Burden; Engineering; Environment; Etiology; Evaluation; Event; Extravasation; Fibrosis; Flare; Foundations; Frequencies; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genetic Transcription; Goals; Grant; Health; Home; Human; Immune; In Vitro; Individual; Infection; Inflammation; Inflammatory; Laboratories; Link; Liquid substance; Long COVID; Longitudinal Studies; Malignant Female Reproductive System Neoplasm; Measurement; Medical; Mental disorders; Methods; Microfluidics; Modeling; Molecular; Molecular Profiling; Neurologic; Paracrine Communication; Parents; Participant; Partner in relationship; Pathway interactions; Patients; Pattern; Persons; Phase; Pilot Projects; Population; Positioning Attribute; Post-Acute Sequelae of SARS-CoV-2 Infection; Production; RNA; RNA analysis; Recurrence; Research; Resolution; Role; Sample Size; Sampling; Schedule; Shipping; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Therapeutic; Time; Transportation; Tube; United States National Institutes of Health; Uterine Cancer; Vasodilation; Vocabulary; Woman; Work; analytical method; base; caregiving; clinical research site; cohort; experience; gender difference; individual variation; insight; laboratory experience; longitudinal analysis; malignant breast neoplasm; member; men; novel; peripheral blood; preservation; pressure; prevent; research study; response; rural area; small molecule; social; therapeutic target; tool; transcriptomics; treatment response; trend; vaccine development

Project Abstract Small molecule and protein signals provide a rich vocabulary for cellular communication, and upstream changes in RNA expression help drive the molecular dialogue. The downstream consequences of gene expression changes and signal molecule production are exquisitely sensitive and vary based on microenvironment, disease state (including temporal considerations), and individual heterogeneity. For example, gene expression changes vary temporally in response to disease flares or treatments. Dissecting the basic biology of cell signaling and molecular pathway changes in disease is challenging, and new methods are required to address fundamental questions: What is the downstream biological function of each signaling molecule? How is the biological function different when molecules are present in mixtures? How do signaling pathways vary across the human population and through time within an individual? To address these challenges, the parent R35 award has three stated goals: 1) Develop novel microscale co- and multi-culture platforms to study soluble factor signaling and use these tools to elucidate paracrine signaling mechanisms. 2) Develop and validate new readouts for inflammation (e.g., fibrosis, vasodilation) and apply these methods to identify key effector molecules and signaling pathways in inflammation. 3) Develop new analytical methods to stabilize, isolate, and study inflammatory signals. Under the parent R35 award Goal 3, we developed a novel platform that enables at-home blood sampling and RNA stabilization, homeRNA. The homeRNA kit contains a commercially available at-home blood collection device and a custom ‘stabilizer tube’ that our lab engineered to contain a stabilizing solution (e.g., RNAlater to stabilize RNA in blood). Leveraging our lab’s experience with microfluidics and utilizing passive forces (e.g., capillary flow, Laplace pressure), we engineered the ‘stabilizer tube’ with an integrated fluidic channel that prevents spillage of RNAlater while the user handles the kit, but enables the transfer of fluid (e.g., RNAlater solution) when the stabilizer tube is mated with the blood collection tube. Importantly, homeRNA enables longitudinal studies within an individual to capture temporal changes in gene expression signatures resulting from disease flares or in response to treatment. homeRNA enables evaluation of mechanistic hypotheses in human populations, providing a complement to our lab’s in vitro microfluidic platforms, which use cell lines or primary cells. In this supplement, we will use homeRNA to study the molecular mechanisms underlying inflammation in women of understudied, underrepresented, and underreported (U3) populations experiencing post-acute sequelae of SARS-CoV-2 (PASC, also called ‘long COVID’), ultimately enabling better diagnostic and therapeutic approaches for PASC. Further, we will establish homeRNA as a broadly applicable research tool for reaching women of U3 populations.

项目摘要 小分子和蛋白质信号为细胞通信提供了丰富的词汇，上游变化 在RNA表达中有助于推动分子对话。基因表达的下游后果 变化和信号分子的产生完全敏感，并且根据微环境，疾病而变化 状态（包括临时考虑）和个体异质性。例如，基因表达改变 暂时响应疾病或治疗而有所不同。剖析细胞信号的基本生物学和 疾病的分子途径变化具有挑战性，需要新的方法来解决基本 问题：每个信号分子的下游生物学功能是什么？生物功能如何 混合物中存在分子时有所不同吗？信号通路如何在人口中变化 在一个人内的时间？为了应对这些挑战，父母R35奖已有三个 目标：1）开发新型的微观合作和多元文化平台来研究固体因素信号并使用这些平台 阐明旁分泌信号传导机制的工具。 2）开发和验证新的炎症读数（例如， 纤维化，血管舒张）并应用这些方法来识别关键效应分子和信号通路 炎。 3）开发新的分析方法来稳定，分离和研究炎症信号。在 家长R35奖项目标3，我们开发了一个新颖的平台，可实现在家取样和RNA 稳定，Homerna。 HOMERNA套件包含一个可商购的在家血液收集装置 以及我们设计的定制“稳定管”，该实验室设计为稳定溶液（例如，稳定剂量 血液中的RNA）。利用我们实验室在微流体学和使用被动力量的经验（例如，毛细管流， 拉普拉斯压力），我们用集成的流体通道设计了“稳定管”，以防止溢出 当用户处理套件时，rnalater，但在启用流体转移（例如，rnalater溶液） 稳定管是与血液收集管的伴侣。重要的是，Homerna可以在 一个人捕获疾病耀斑或中引起的基因表达特征的暂时变化 对治疗的反应。 Homerna可以评估人类种群的机理假设， 为我们的实验室体外微流体平台提供了完善，该平台使用细胞系或主要细胞。在这个 补充，我们将使用Homerna研究女性的分子机制 代表性不足，报道不足的（U3）种群经历了急性后遗症的后遗症 SARS-COV-2（PASC，也称为“ Long covid”），最终使能够更好地诊断和治疗 PASC的方法。此外，我们将建立Homerna作为一种广泛适用的研究工具 U3人口的妇女。