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

这个PFI项目的更广泛的影响/商业潜力将来自开发有效和均匀地将分子输送到细胞内部的新设备。许多大的或带电的分子不容易穿透细胞的外层，也不能接触到细胞内的成分。这是一个问题，因为将某些分子(包括糖、蛋白质、DNA或RNA)输送到细胞内部将使修改和储存细胞的方法变得更好。这可能有用的例子包括一种受自然启发的制备冷冻干燥细胞的过程，以便它们可以在室温下储存并根据需要进行再水化，这将对研究(细胞系)和医疗(红细胞)都有用。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健康和支持国防。该项目还旨在加强学术界和产业界之间的关系，并鼓励妇女和代表不足群体的个人参与STEM研究。拟议的项目预计将促进许多领域的知识，包括超声波介导声手术、微流体和低温生物学。我们的最终目标是开发集成的超声-微流体设备，以便最佳地将细胞不渗透分子输送到各种类型的细胞。目前的方法并不理想，原因有很多，包括低的转染率和毒性。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)输送到红细胞或培养细胞系，使细胞能够被冷冻、脱水、在常温下储存，然后再水化，而不会造成大量活性或功能损失；以及(2)将生物分子(DNA、RNA或蛋白质)输送到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控设计、流速、冷冻保存或转染液，以及声孔、冷冻、干燥和回收的方法。我们方法的成功将通过评估多个因素来决定，例如负载、生存能力和功能。这是一个学术界和业界合作的项目，大学研究人员和行业专家将共同努力，最大限度地提高开发速度和项目产生商业可行产品的可能性。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。

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.