Single-Molecule Imaging of Biological Trauma: Cytokine-Based Intracellular Communication

生物创伤的单分子成像：基于细胞因子的细胞内通讯

• 批准号: 10176532
• 负责人: Markita Landry
• 金额: $ 36.25万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176532
• 关键词: Address; Appointment; Area; Autoimmune Diseases; Award; Biological; Biological Markers; Biophysics; Biopsy; Cells; Central Nervous System Diseases; Chemical Engineering; Chemicals; Clinical; Communication; Cultured Cells; Cytokine Signaling; Detection; Development; Diagnostic; Disease; Environment; Epithelial Cells; Faculty; Fluorescence; Goals; IL8 gene; Immune; In Vitro; Individual; Infection; Interleukin-6; Laboratory Research; Language; Malignant Neoplasms; Measurement; Measures; Methods; Microscopy; Molecular; Monitor; Nanotechnology; Nature; Optics; Peptide Signal Sequences; Plasma; Positioning Attribute; Postdoctoral Fellow; Research; Sampling; Science; Serum; Signal Transduction; Spectrum Analysis; Time; Training; Translating; Trauma; Variant; Vascular Endothelial Growth Factors; Work; base; chemokine; cytokine; design; insight; instrument; macrophage; molecular imaging; molecular recognition; nanomaterials; nanoscale; nanosensors; personalized diagnostics; programs; sensor; single molecule; spatiotemporal; synergism; targeted treatment; tool; tumor

PROJECT ABSTRACT Research Abstract: The detection of unlabeled cytokines in real-time and from single cells could provide a robust platform for understanding the ‘molecular language of biological trauma and disease’. However, tools to visualize cytokines at the cellular level, particularly in their secreted form, are lacking. We have developed a generic nanomaterial-based near-infrared fluorescent sensor and accompanying microscopy platform which produces a unique intensity and wavelength shift in the presence of a specific target molecule (Zhang*, Landry* et al. Nature Nanotechnology 2013; Landry et al. Sensors 2015; Landry et al. Nature Nanotechnology 2017). In this 5-year proposal, I (i) will develop synthetic sensors for VEGF, IL-6, and IL-8 cytokines, (ii) validate their use to monitor constitutive cytokine secretion from macrophage and epithelial cells, and (iii) directly visualize the spatio-temporal profiles of intercellular cytokine-based synergies. Direct cellular measurement of secreted cytokines will inform how cytokine secretion profiles from single or few individual cells are stimulated by chemokines and cytokines, which forms the basis of the cytokine secretion profiles currently used in biomarker- based diagnostics. The research we propose herein has – to the best of our knowledge – only been explored theoretically (Thurley et al. POLS Comp. Bio. 2015). Landry Laboratory Research Program: I am a single-molecule biophysicist by training, having developed several instruments capable of detecting piconewton-scale forces (Landry et al. Biophys. J. 2009), and nanometer-scale fluorescence localization (Landry et al. Nucl. Ac. Res. 2012) for my doctoral work. In transitioning to my postdoctoral position, my goal was to leverage my expertise in single-molecule spectroscopy and molecular biophysics to design purely synthetic molecular recognition tools. My scientific training in as a postdoctoral fellow in Chemical Engineering at MIT focused on merging these two previously disparate areas of science: optical microscopy and nanosensor development, yielding a platform for the optical detection of any generic molecular analyte. I began my faculty appointment at UC Berkeley in June 2016, with a research portfolio motivated by translating the technical strengths of my lab in microscopy (O’Donnel et al. Adv. Funct. Mater 2017), sensor development (Beyene et al. ACS Chem Neruo 2017 & Luo et al. ACS Sensors 2017), and molecular recognition (Li et al. RSC Chemical Science 2017) to addressing the need to develop methods to detect cytokine efflux from immune cells. In the first two years of my research plan, my group will synthesize and characterize nanomaterial-based sensors for cytokines in vitro. The remaining three years of the R35 award will implement the use of cytokine sensors to measure constitutive (year 3), induced (year 4), and intracellular (year 5) cytokine signaling from cultured cell samples. My long-term research goals focus on the application of nanosensors for cytokines, chemokines, and other important biomarkers in environments such as multicellular tumor spheroids and live tumor biopsies, in which biomarker detection has traditionally been difficult. Landry ABSTRACT AB-1

项目摘要 研究摘要：实时从单个细胞中检测未标记的细胞因子可以提供一种 理解“生物创伤和疾病的分子语言”的强大平台。然而，工具可用于 缺乏细胞水平的可视化细胞因子，特别是分泌形式的细胞因子。我们已经开发出一种 一种通用纳米材料近红外荧光传感器及伴随显微镜平台， 在特定目标分子(张*、兰德里*)存在的情况下产生独特的强度和波长移动 等人的研究。自然纳米技术，2013；Landry等人。传感器，2015；Landry等人。自然纳米技术2017年)。在……里面 这项为期5年的计划，I(I)将开发用于VEGF、IL-6和IL-8细胞因子的合成传感器，(Ii)验证它们的用途 监测巨噬细胞和上皮细胞的结构性细胞因子分泌，以及(Iii)直接显示 细胞间细胞因子协同作用的时空分布。分泌物的直接细胞测量 细胞因子将决定单个或少数单个细胞分泌细胞因子的方式。 趋化因子和细胞因子，它们构成了目前用于生物标记物的细胞因子分泌谱的基础- 基于诊断的。据我们所知，我们在这里提出的研究只被探索过 理论上(瑟利等人)。POLS公司生平。2015年)。 兰德里实验室研究计划：我是一名经过培训的单分子生物物理学家，已经开发出 几种能够探测皮牛顿力的仪器(Landry等人)。生物群落。J.2009)，以及 纳米尺度荧光定位(Landry等人。核子。交流。2012)，以感谢我的博士工作。在……里面 过渡到我的博士后职位，我的目标是利用我在单分子光谱学方面的专业知识 和分子生物物理学来设计纯合成的分子识别工具。我的科学训练是作为 麻省理工学院化学工程博士后专注于合并这两个以前不同的领域 科学：光学显微镜和纳米传感器的发展，为光学检测任何 普通分子分析物。2016年6月，我开始在加州大学伯克利分校担任教职，带着一个研究文件 动机是将我的实验室在显微镜方面的技术优势转化为(O‘Donnel等人)。Adv.Funct.2017年)， 传感器开发(Beyene等人ACSChem Neruo 2017&Luo et al.ACS传感器2017)和分子 认可(Li等人)RSC化学科学2017)以解决开发检测细胞因子的方法的需求 从免疫细胞外流。在我的研究计划的头两年，我的团队将综合和描述 基于纳米材料的体外细胞因子传感器。R35奖励的剩余三年将实施 使用细胞因子传感器测量构成(第3年)、诱导(第4年)和细胞内(第5年)细胞因子 来自培养细胞样本的信号。我的长期研究目标是将纳米传感器应用于 多细胞肿瘤球体等环境中的细胞因子、趋化因子和其他重要生物标志物 活体肿瘤活组织检查，其中生物标记物的检测传统上是困难的。 兰德里摘要AB-1