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.

工程纳米材料已经在诊断、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的且相对未知的，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。这个CAREER奖将资助一个研究项目，该项目引入了一种新的成像技术，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细胞健康的既定措施相关联，从而更好地了解纳米材料诱导的毒性。这项工作将紧密结合到一个教育和推广计划，使当地的K-12社区参与互动研讨会和动手实验室经验调查在纳米生物界面的相互作用。最后，这项工作将能够形成一个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得岛内以STEM为重点的劳动力。总之，这些活动通过产生对STEM的广泛兴趣和提高对新型纳米材料技术的理解，支持工作的更广泛影响和传播。在过去的二十年里，工程纳米材料用于创造新的诊断，成像和药物输送设备已经获得了极大的关注。最近发现的碳的一维同素异形体，单壁碳纳米管，具有内在的近红外荧光，是无限的光稳定性和环境敏感性，提供了一个独特的机会，创造传感和成像结构作为活细胞和动物研究的研究工具。与所有外源引入的材料一样，应彻底探讨其对细胞健康的短期和长期不利影响。在碳纳米管的情况下，已知纳米材料通过受体介导的内吞作用进入细胞，并保持在内体途径内。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，应该研究对参与该途径的囊泡的有害影响。该CAREER项目的研究目标是使用一种新的光谱成像方法进行活细胞内的亚细胞测量，以量化细胞内衍射限制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来研究具有不同物理性质的工程化纳米管在生物介质中表现出的纳米稳定性，2）研究纳米管官能化、浓度和聚集状态如何影响摄取和内体负载率（每个内体的纳米管），以及3）检查内体装载率如何影响天然内体成熟过程（囊泡运输、内体至溶酶体进展等），并将其与各种细胞应激和毒性测定相关联。这项工作的变革性源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，这种方式不需要标记或干扰手头的生物系统。通过执行这些测定，将创建一个规则框架，该框架管理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，这项工作将使一个高度跨学科的生物纳米技术课程的形成，突出纳米毒理学的主题，以进一步刺激和教育罗得岛内的STEM为重点的劳动力。这个奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

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