CAREER: Spectral imaging for sub-cellular nanometrology and nanotoxicology

职业：亚细胞纳米计量学和纳米毒理学的光谱成像

• 批准号: 1844536
• 负责人: Daniel Roxbury
• 金额: $ 50万
• 依托单位: University of Rhode Island
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1844536&HistoricalAwards=false
• 关键词: CAREER; Spectral; imaging; sub; cellular

Engineered nanomaterials have demonstrated biomedical applications in diagnostics, imaging, and drug delivery. For widespread adoption of these technologies, potential adverse health effects in both the short and long-term should be thoroughly explored. Due to their novel nanoscale properties, the manner in which these nanomaterials interact with biological systems is complex and relatively unknown, and very few tools exist to accurately understand the fate of these nanomaterials in cells, animals and humans. This CAREER award will fund a research program that introduces a new imaging technique to more precisely describe the locations and numbers of nanomaterials within the confines of live cells. This research will generate new knowledge, where quantitative imaging data will be correlated to established measures of cell health, leading to a better understanding of nanomaterial-induced toxicity. The work will be closely integrated into an educational and outreach program that engages the local K-12 community with interactive seminars and hands-on laboratory experience investigating interactions at the nano-bio interface. Finally, the work will enable the formation of a highly interdisciplinary bionanotechnology course, with particular emphasis on nanotoxicology, to further stimulate and educate the STEM-focused workforce within Rhode Island. Together, these activities support the broader impacts and dissemination of the work by generating widespread interest in STEM and improved understanding of novel nanomaterial technologies. Engineering nanomaterials for the purposes of creating novel diagnostic, imaging, and drug delivery devices has garnered significant attention within the past two decades. A recently discovered one-dimensional allotrope of carbon, the single-walled carbon nanotube, with intrinsic near-infrared fluorescence that is indefinitely photostable and environmentally sensitive, presents a unique opportunity to create sensing and imaging constructs as research tools for live cell and animal studies. As with all exogenously introduced materials, adverse effects to cell health in both the short and long-term should be thoroughly explored. In the case of carbon nanotubes, the nanomaterial is known to enter cells through receptor-mediated endocytosis and remain within the endosomal pathway. For widespread adoption of nanotube-based sensors and imaging probes in standard biological applications, detrimental effects to the vesicles involved in this pathway should be investigated. The research objective of this CAREER project, using a novel spectral imaging approach for sub-cellular measurements within live cells, is to quantify the number of nanotubes in diffraction-limited regions within a cell and determine how naturally aggregated nanomaterials influence toxicity in mammalian cells. The research project seeks to: 1) employ hyperspectral fluorescence microscopy to investigate the nanostability that engineered nanotubes of varying physical properties exhibit in biological media, 2) investigate how nanotube functionalization, concentration, and aggregation state can affect the uptake and endosome loading ratio (nanotubes per endosome) in mammalian cells, and 3) examine how endosome loading ratio influences natural endosomal maturation processes (vesicle trafficking, endosome-to-lysosome progression, etc.), and correlate this to various cell stress and toxicity assays. The transformative nature of this work stems from the ability to pose and answer outstanding questions of nanotoxicology at the single-cell and sub-cellular level in a new manner that does not necessitate labeling or perturbing the biological system at hand. By performing these assays, a framework of rules will be created that governs the manner in which mammalian cells interact with nanotubes of various physical natures. The work will be thoroughly incorporated into a K-12 educational and outreach program with interactive seminars and practical laboratory experience probing biophysical interactions at the nano-bio interface. Finally, the work will enable the formation of a highly interdisciplinary bionanotechnology course, highlighting topics in nanotoxicology, to further stimulate and educate the STEM-focused workforce within Rhode Island.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.

工程纳米材料已在诊断、成像和药物输送方面展示了生物医学应用。为了广泛采用这些技术，应彻底探讨短期和长期潜在的不利健康影响。 由于其新颖的纳米级特性，这些纳米材料与生物系统相互作用的方式是复杂且相对未知的，并且很少有工具可以准确了解这些纳米材料在细胞、动物和人类中的命运。该职业奖将资助一项研究计划，该计划引入一种新的成像技术，以更精确地描述活细胞范围内纳米材料的位置和数量。这项研究将产生新的知识，其中定量成像数据将与既定的细胞健康指标相关联，从而更好地了解纳米材料引起的毒性。这项工作将紧密地融入到一项教育和推广计划中，该计划让当地 K-12 社区参与互动研讨会并获得研究纳米生物界面相互作用的实验室实践经验。最后，这项工作将有助于形成高度跨学科的生物纳米技术课程，特别强调纳米毒理学，以进一步刺激和教育罗德岛州内以 STEM 为重点的劳动力。这些活动共同激发了人们对 STEM 的广泛兴趣并提高了对新型纳米材料技术的理解，从而支持了更广泛的影响和传播工作。在过去的二十年里，用于制造新型诊断、成像和药物输送设备的工程纳米材料引起了人们的广泛关注。最近发现的碳的一维同素异形体，即单壁碳纳米管，具有无限光稳定性和环境敏感性的固有近红外荧光，为创建传感和成像结构作为活细胞和动物研究的研究工具提供了独特的机会。与所有外源引入的材料一样，应彻底探讨短期和长期对细胞健康的不利影响。就碳纳米管而言，已知纳米材料通过受体介导的内吞作用进入细胞并保留在内体途径内。为了在标准生物应用中广泛采用基于纳米管的传感器和成像探针，应该研究对该途径所涉及的囊泡的有害影响。该 CAREER 项目的研究目标是使用一种新颖的光谱成像方法进行活细胞内的亚细胞测量，量化细胞内衍射极限区域中的纳米管数量，并确定自然聚集的纳米材料如何影响哺乳动物细胞的毒性。该研究项目旨在：1) 采用高光谱荧光显微镜研究不同物理性质的工程纳米管在生物介质中表现出的纳米稳定性，2) 研究纳米管功能化、浓度和聚集状态如何影响哺乳动物细胞中的摄取和内体装载率（每个内体的纳米管），以及 3) 检查内体装载率如何影响自然 内体成熟过程（囊泡运输、内体到溶酶体的进展等），并将其与各种细胞应激和毒性测定相关联。这项工作的变革性本质在于能够以一种新的方式在单细胞和亚细胞水平上提出和回答纳米毒理学的突出问题，而无需标记或扰乱现有的生物系统。通过进行这些测定，将创建一个规则框架，控制哺乳动物细胞与各种物理性质的纳米管相互作用的方式。这项工作将被彻底纳入 K-12 教育和推广计划，其中包括互动研讨会和探索纳米生物界面生物物理相互作用的实际实验室经验。最后，这项工作将有助于形成高度跨学科的生物纳米技术课程，突出纳米毒理学主题，以进一步刺激和教育罗德岛州内以 STEM 为重点的劳动力。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优点和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Enhancing the Thermal Stability of Carbon Nanomaterials with DNA

利用 DNA 增强碳纳米材料的热稳定性

• DOI: 10.1038/s41598-019-48449-x
• 发表时间: 2019
• 期刊: Scientific Reports
• 影响因子: 4.6
• 作者: Safaee, Mohammad Moein;Gravely, Mitchell;Lamothe, Adeline;McSweeney, Megan;Roxbury, Daniel
• 通讯作者: Roxbury, Daniel

A Wearable Optical Microfibrous Biomaterial with Encapsulated Nanosensors Enables Wireless Monitoring of Oxidative Stress

• DOI: 10.1002/adfm.202006254
• 发表时间: 2021-01-15
• 期刊: ADVANCED FUNCTIONAL MATERIALS
• 影响因子: 19
• 作者: Safaee, Mohammad Moein;Gravely, Mitchell;Roxbury, Daniel
• 通讯作者: Roxbury, Daniel

Aggregation Reduces Subcellular Localization and Cytotoxicity of Single-Walled Carbon Nanotubes

• DOI: 10.1021/acsami.2c02238
• 发表时间: 2022-05-04
• 期刊: ACS APPLIED MATERIALS & INTERFACES
• 影响因子: 9.5
• 作者: Gravely, Mitchell;Kindopp, Aidan;Roxbury, Daniel
• 通讯作者: Roxbury, Daniel

Multispectral Fingerprinting Resolves Dynamics of Nanomaterial Trafficking in Primary Endothelial Cells

• DOI: 10.1021/acsnano.1c04500
• 发表时间: 2021-06-28
• 期刊: ACS NANO
• 影响因子: 17.1
• 作者: Gravely, Mitchell;Roxbury, Daniel
• 通讯作者: Roxbury, Daniel

Biomolecular Functionalization of a Nanomaterial To Control Stability and Retention within Live Cells

• DOI: 10.1021/acs.nanolett.9b02267
• 发表时间: 2019-09-01
• 期刊: NANO LETTERS
• 影响因子: 10.8
• 作者: Gravely, Mitchell;Safaee, Mohammad Moein;Roxbury, Daniel
• 通讯作者: Roxbury, Daniel