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.

工程纳米材料已经展示了在诊断、成像和药物输送方面的生物医学应用。为了广泛采用这些技术，应彻底探索短期和长期对健康的潜在不利影响。由于它们新颖的纳米级特性，这些纳米材料与生物系统相互作用的方式是复杂的，相对未知的，而且几乎没有工具可以准确地了解这些纳米材料在细胞、动物和人类中的命运。这项职业奖将资助一个研究项目，该项目引入了一种新的成像技术，可以更准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测量相关联，从而更好地理解纳米材料诱导的毒性。这项工作将被紧密地整合到一个教育和外展计划中，该计划通过互动研讨会和实际实验室经验调查纳米生物界面上的相互作用，使当地的K-12社区参与进来。最后，这项工作将有助于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德岛以STEM为重点的劳动力。总而言之，这些活动通过引起人们对STEM的广泛兴趣和增进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，为了创造新的诊断、成像和药物输送装置而设计的纳米材料引起了人们的极大关注。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，具有无限的光稳定性和环境敏感性，为创造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。与所有外源材料一样，对细胞健康的短期和长期不利影响都应得到彻底的研究。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，并留在内体途径中。为了在标准的生物学应用中广泛采用基于纳米管的传感器和成像探针，应该调查这一途径中涉及的囊泡的有害影响。这个职业项目的研究目标是利用一种新的光谱成像方法在活细胞内进行亚细胞测量，以量化细胞内衍射受限区域中的纳米管的数量，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)使用高光谱荧光显微镜来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)研究纳米管的功能化、浓度和聚集状态如何影响哺乳动物细胞的摄取和内小体负载率(每个内小体的纳米管)，以及3)检测内小体负载率如何影响自然的内小体成熟过程(囊泡运输，内小体到溶酶体的进展等)，并将其与各种细胞压力和毒性分析相关联。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提出和回答悬而未决的纳米毒理学问题，而不需要标记或扰乱手头的生物系统。通过进行这些分析，将建立一个规则框架，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，通过互动研讨会和实际实验室经验探索纳米生物界面上的生物物理相互作用。最后，这项工作将有助于形成一个高度跨学科的生物纳米技术课程，突出纳米毒理学的主题，以进一步激励和教育罗德岛以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势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