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

这个小企业创新研究（SBIR）第二阶段项目的更广泛的影响/商业潜力将是开发将生物分子直接输送到细胞内的技术。这种基于微流体的平台有可能成为细胞内递送的一种使能技术，它可以通过允许可靠、有效地递送各种材料类别而不必设计材料或细胞来天然摄取这些分子来加速药物发现研发。这种能力可以使制药公司比以往任何时候都更快地评估候选药物的疗效，特别是与已经完善和有效的高通量机器人工作流程集成。该技术可以通过将候选药物的活性与细胞对分子的亲和力分离来大大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理解。与领先的药物开发商和学术实验室朝着这一目标进行的初步研究非常令人鼓舞，并且在未来，该平台可能能够对细胞功能进行强大的工程改造，用于针对多种疾病的基于细胞的疗法，包括流感，癌症，甚至自身免疫性疾病。该SBIR第二阶段项目提出了细胞内递送技术的持续发展，以解决药物发现&研发新药物的发现通常受到膜不可渗透的候选药物无法进入细胞胞质溶胶的阻碍，需要外源性材料进行递送，如强电场或病毒载体。然而，这些材料往往会引起脱靶效应或毒性，因此需要一种可以促进递送而不改变治疗后细胞功能的技术。该项目的目标是展示一个面向市场采用微流体硬件作为转染和细胞内递送标准方法的平台。在第二阶段，该平台将进行充分的特征描述、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，研究计划通过开发CRISPR/Cas9基因编辑系统与这种细胞内递送技术一起使用来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.