SBIR Phase II: Development of an Intracellular Delivery Platform for Accelerated Drug Discovery Using Genetically Engineered Human Immune Cells

这一小型企业创新研究(SBIR)第二阶段项目的更广泛影响/商业潜力将是开发细胞内生物分子直接进入细胞的技术。这种基于微流体的平台有可能成为一种细胞内传递的使能技术，通过允许可靠、高效地传递不同类型的材料，而不必设计材料或细胞来天然地吸收这些分子，这可能被用于加速药物发现研发。这种能力可以让制药公司比以往任何时候都更快地评估候选药物的疗效，特别是在集成到已经建立良好和有效的高通量机器人工作流程的情况下。这项技术可以将候选药物活性的确定与细胞对分子的亲和力分离，从而极大地缩短新药上市的时间。它还可以促进对生物过程和途径的更深入了解。与领先的药物开发商和学术实验室为实现这一目标进行的初步研究非常令人鼓舞，在未来，该平台可能会为针对包括流感、癌症甚至自身免疫性疾病在内的各种疾病的细胞疗法实现强大的细胞功能工程。这个SBIR第二阶段项目建议继续开发细胞内递送技术，以满足药物研发中的相关应用。新药发现经常受到膜不透性候选药物无法进入细胞胞浆的阻碍，需要使用外源材料进行递送，例如强电场或病毒载体。然而，这些材料往往会引起非靶标效应或毒性，这就需要一种技术，在不改变治疗后细胞功能的情况下促进递送。该项目的目标是展示一个面向市场的平台，该平台采用微流控硬件作为转基因和细胞内传递的标准方法。在第二阶段，该平台将经过充分的表征、验证和验证，以产生实现市场进入所需的一致、可重复的结果。此外，计划进行研究，通过开发与这种细胞内递送技术一起使用的CRISPR/Cas9基因编辑系统，证明该平台支持药物发现研发的能力。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project will be the development of technology for the intracellular delivery of biomolecules directly into cells. This microfluidics-based platform has the potential to become an enabling technology for intracellular delivery, which may be used to accelerate drug discovery R&D by allowing reliable, efficient delivery of diverse material classes without having to engineer the material or the cell to natively uptake these molecules. Such capabilities could allow pharmaceutical companies to assess the efficacy of drug candidates faster than ever before, especially with integration into high-throughput robotic workflows that are already well-established and efficacious. The technology could dramatically reduce the time to market for new drugs by decoupling determination of a candidate's activity from the cell's affinity for the molecule. It also could facilitate a deeper understanding of biological processes and pathways. Initial studies with leading drug developers and academic laboratories towards this goal have been very encouraging, and, in the future, the platform could potentially enable robust engineering of cell function for cell-based therapies targeting a diversity of diseases including influenza, cancer, and even autoimmune disorders.This SBIR Phase II project proposes the continued development of the intracellular delivery technology to address relevant applications in drug discovery R&D. New drug discovery is often hampered by the inability of membrane-impermeable drug candidates to enter the cell cytosol, necessitating exogenous materials for delivery such as strong electric fields or viral vectors. However, these materials tend to cause off-target effects or toxicity, presenting a need for a technology that can facilitate delivery without altering post-treatment cellular function. The goal of this project is to demonstrate a platform geared towards market adoption of microfluidic hardware as the standard method for transfection and intracellular delivery. During Phase II, the platform will be fully-characterized, validated, and verified in order to produce the consistent, repeatable results necessary to achieve market entry. In addition, research is planned to demonstrate the ability of the platform to support drug discovery R&D by developing the use of the CRISPR/Cas9 gene editing system for use with this intracellular delivery technology.