SBIR Phase I: An Automated Microfluidic Platform for Delivery of Biomolecules Into Cells

这个小型企业创新研究(SBIR)项目的更广泛的影响/商业潜力是解决基础生物学研究和下一代临床治疗中的一个主要障碍：将材料输送到细胞中。细胞是人体的基本功能单位，然而，由于我们无法将材料输送到细胞质，因此了解它们在疾病中的作用并利用它们固有的潜力来抗击疾病一直受到限制。通过促进进入细胞内部，可以使探测细胞内过程和为治疗目的设计细胞功能的能力迅速进步。该项目旨在进一步开发一种有前景的细胞内递送的新概念，能够克服与当前最先进技术相关的许多传统障碍。该平台将潜在地促进基于对细胞功能的更深入了解和更强大的设计细胞命运的能力的新疗法的开发。事实上，解决生物医学领域的这一根本挑战将为社会带来实质性的好处，并可能影响许多商业机会。潜在的应用包括基础研究、高通量药物发现筛选和基于细胞的治疗癌症免疫疗法。这个SBIR第一阶段项目建议开发一种无载体的微流控平台，用于细胞内生物分子的传递，以提高效率，并改善易用性。该平台使用了一种新的方法，该方法基于细胞通过微流控收缩时的快速、瞬时变形(细胞挤压)。挤压过程导致细胞膜的暂时破坏，并促进靶向递送材料的被动运输到细胞质中。这项拟议的工作旨在引入对控制输送过程的关键参数(压力、温度和流速)的自动化、闭环控制。这些添加将允许用户精确地调整输送到细胞的材料的量和由此产生的生存能力。通过开发这一硬件，到第一阶段结束时，这项技术将为更多的采用和商业化奠定良好的基础。建议的硬件控制器将通过使用初级免疫细胞的相关研究进行验证和验证。初级免疫细胞是与疾病相关的细胞子集，与现有的输送方法不相容。最后，拟议的工作将有助于推出一个强大的原型系统，用于高影响应用程序的早期测试。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is to address a major barrier in fundamental biological research and next-generation clinical treatments: Delivering materials into cells. Cells are the basic functional unit of the body yet understanding their role in disease and harnessing their inherent potential to combat ailments has been limited by our inability to deliver material to their cytoplasm. By facilitating access to a cell's interior one could enable rapid progress in the ability to probe intracellular processes and engineer cell function for therapeutic purposes. This project aims to further develop a promising new concept of intracellular delivery capable of overcoming many conventional barriers associated with the current state-of-the-art. The platform will potentially facilitate the development of novel therapeutics based on a deeper understanding of cell function and a more robust ability to engineer cell fate. Indeed, addressing such a fundamental challenge in the biomedical field would provide substantial benefits to society and could impact numerous commercial opportunities. Potential applications include basic research, high-throughput drug discovery screening, and cell-based therapies to treat cancer immunotherapies. This SBIR Phase I project proposes to develop a vector-free microfluidic platform for intracellular delivery of biomolecules in order to increase efficacy, and improve ease-of-use. The platform uses a novel method based on rapid, transient deformation of cells ("cell squeezing") as they pass through a microfluidic constriction. The squeezing process causes temporary disruption of the cell membrane and facilitates passive transport of target delivery materials into the cytoplasm. The proposed work aims to introduce automated, closed-loop control of key parameters (pressure, temperature, and flow rate) that govern the delivery process. These additions will allow users to precisely tune the amount of material delivered to cells and the resultant viability. By developing this hardware, the technology will be well-positioned for increased adoption and commercialization by the end of Phase I. The proposed hardware controllers will be verified and validated through relevant studies using primary immune cells, a disease-relevant subset of cells that are recalcitrant to existing delivery methods. Finally, the proposed work would facilitate the launch of a robust prototype system for early-stage testing in high-impact applications.