PFI-RP: Development of Delivery Devices to Enable Cell Transformation or Preservation

PFI-RP：开发递送装置以实现细胞转化或保存

• 批准号: 1827521
• 负责人: Jonathan Kopechek
• 金额: $ 75万
• 依托单位: University of Louisville Research Foundation Inc
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2023-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1827521&HistoricalAwards=false
• 关键词: PFI; RP; Development; Delivery; Devices

The broader impact/commercial potential of this PFI project will arise from the development of new devices that efficiently and uniformly transport molecules to the inside of cells. Many large or charged molecules do not easily penetrate the outer layer of cells and are not able to access the components inside the cell. This is a problem because delivery of certain molecules (including sugars, proteins, DNA, or RNA) to the inside of cells would allow better methods for modifying and storing cells. Examples of how this could be useful include a nature-inspired process for preparing freeze-dried cells so they can be stored at room temperature and rehydrated on demand, which would be useful for both research (cell lines) and medical treatments (red blood cells). If successful, these activities will increase the economic competitiveness of the United States through new product development, company formation, and training opportunities. Our research could advance public health and support national defense by improving cancer therapy and providing dried blood for military field use, space travel, and emergency preparedness. This project is also designed specifically to enhance relationships between academia and industry, and to encourage participation of women and individuals from underrepresented groups in STEM research.The proposed project is expected to advance knowledge in many areas, including ultrasound-mediated sonoporation, microfluidics, and cryobiology. Our ultimate goal is to develop integrated ultrasound-microfluidic devices for optimal delivery of cell-impermeable molecules to a variety of cell types. Current methods are not ideal for numerous reasons including low transfection efficiency and toxicity. We plan to develop devices that can be used for the following applications: (1) intracellular delivery of cryoprotectant molecules (such as trehalose) to red blood cells or to cultured cell lines so that cells can be frozen, dehydrated, stored at ambient temperature, and then rehydrated without substantial loss of viability or function; and (2) intracellular delivery of biomolecules (DNA, RNA, or protein) to cultured cells or primary cells for use in research or therapy. Parameters to be optimized include microfluidic designs, flow rate, cryopreservation or transfection solutions, and methods for sonoporating, freezing, drying, and recovery. The success of our approach will be determined by evaluating multiple factors, such as loading, viability, and function. This is an academia-industry partnership project where university researchers and industry experts will work together to maximize the speed of development and the likelihood that the project will result in a commercially viable product.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.

这个PFI项目更广泛的影响/商业潜力将来自于新设备的开发，这些设备可以高效、均匀地将分子输送到细胞内部。许多大分子或带电分子不容易穿透细胞的外层，也不能接触到细胞内部的成分。这是一个问题，因为将某些分子（包括糖、蛋白质、DNA或RNA）运送到细胞内部将允许有更好的方法来修饰和储存细胞。这可能有用的例子包括一个受自然启发的制备冻干细胞的过程，这样它们就可以在室温下储存并根据需要再水化，这对研究（细胞系）和医学治疗（红细胞）都很有用。如果成功，这些活动将通过新产品开发、公司组建和培训机会增加美国的经济竞争力。我们的研究可以通过改进癌症治疗和为军事野战、太空旅行和应急准备提供干血来促进公共卫生和支持国防。该项目还专门设计用于加强学术界和工业界之间的关系，并鼓励女性和来自代表性不足群体的个人参与STEM研究。该计划预计将推动许多领域的知识，包括超声介导的超声穿孔，微流体和低温生物学。我们的最终目标是开发集成超声微流体装置，以最佳地将细胞不渗透分子输送到各种细胞类型。由于转染效率低和毒性等原因，目前的方法并不理想。我们计划开发可用于以下应用的设备：(1)将冷冻保护剂分子（如海藻糖）细胞内递送到红细胞或培养的细胞系中，使细胞可以冷冻，脱水，在环境温度下储存，然后再水化，而不会严重丧失活力或功能；(2)细胞内递送生物分子（DNA、RNA或蛋白质）至培养细胞或原代细胞，用于研究或治疗。需要优化的参数包括微流体设计、流速、冷冻保存或转染溶液，以及超声、冷冻、干燥和回收的方法。我们的方法的成功将取决于评估多种因素，如负载、生存能力和功能。这是一个学术界和工业界的合作项目，大学研究人员和工业界专家将共同努力，最大限度地提高开发速度，并使项目成为商业上可行的产品。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值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

Assembly and Operation of an Acoustofluidic Device for Enhanced Delivery of Molecular Compounds to Cells

用于增强分子化合物向细胞递送的声流控装置的组装和操作

• DOI: 10.3791/62035
• 发表时间: 2021
• 期刊: Journal of Visualized Experiments
• 作者: Centner, Connor S.;Murphy, Emily M.;Stamp, Bryce F.;Priddy, Mariah C.;Moore, John T.;Bates, Paula J.;Menze, Michael A.;Yaddanapudi, Kavitha;Kopechek, Jonathan A.
• 通讯作者: Kopechek, Jonathan A.

Comparison of Acoustofluidic and Static Systems for Ultrasound-Mediated Molecular Delivery to T Lymphocytes.

• DOI: 10.1016/j.ultrasmedbio.2022.08.005
• 发表时间: 2022-10
• 期刊: Ultrasound in medicine & biology
• 影响因子: 2.9
• 作者: Connor S. Centner;John T Moore;Mary E. Baxter;K. Yaddanapudi;P. Bates;Jonathan A. Kopechek

Acoustofluidic-mediated molecular delivery to human T cells with a three-dimensional-printed flow chamber

• DOI: 10.1121/10.0009054
• 发表时间: 2021-12-01
• 期刊: JOURNAL OF THE ACOUSTICAL SOCIETY OF AMERICA
• 影响因子: 2.4
• 作者: Centner, Connor S.;Moore, John T.;Kopechek, Jonathan A.
• 通讯作者: Kopechek, Jonathan A.

Ultrasound-induced molecular delivery to erythrocytes using a microfluidic system

• DOI: 10.1063/1.5144617
• 发表时间: 2020-03-01
• 期刊: BIOMICROFLUIDICS
• 影响因子: 3.2
• 作者: Centner, Connor S.;Murphy, Emily M.;Kopechek, Jonathan A.
• 通讯作者: Kopechek, Jonathan A.

Rehydration outcomes for freeze-dried red blood cells in reduced gravity

减重状态下冻干红细胞的补液结果

• DOI: 10.1016/j.actaastro.2023.10.006
• 发表时间: 2024
• 期刊: Acta Astronautica
• 影响因子: 3.5
• 作者: Elder, Charles A.;Moore, John;Janis, Brett R.;Shacklette, Sienna;Jones, Clara;Cantrell, Ryan;Grimm, David F.;Pantalos, George;Kopechek, Jonathan A.;Menze, Michael A.
• 通讯作者: Menze, Michael A.