STTR Phase I: Polymer electrosonic actuation microarray: Low-cost tool for transfection of biological cells

这个小型企业技术转移（STTR）第一阶段项目将证明用于生物细胞机电转染的一次性聚合物微阵列的技术和商业可行性。为了获得生物制药、基因治疗和干细胞研究的潜力，需要替代的基因转移解决方案。尽管基因转移的物理方法（如电穿孔）已经证明（与传统的化学介导技术相比）在难以转染的原代细胞和干细胞中的治疗效果有所改善，但治疗的控制和均匀性仍然不足。此外，基于试管的电穿孔系统的性能增量改进只能通过相应的系统复杂性和成本的增加来实现。通过细胞大小的孔，一个接一个的细胞喷射被发现通过在每个细胞上施加相同的和精心控制的机电环境来促进细胞膜穿孔和DNA进入细胞。不幸的是，相对较高的材料成本和可实现的处理条件的限制（由于使用标准硅微加工技术制造的微阵列的几何形状的限制），抑制了现有硅基微阵列的实际实施。在STTR一期项目下，聚合物微阵列的创新设计和优化将产生一种低成本的系统，能够产生所需的机械应力场，以改善难以转染的细胞的治疗效果。该项目更广泛的影响/商业潜力是能够有效和经济地转染难以转染的原代细胞和干细胞，用于生命科学的各种研究和治疗应用。将基因、药物分子、显像剂、多肽、抗体和酶有效地输送到活细胞中，对于从通过引入DNA治疗人类疾病到通过单分子成像研究基本细胞功能的各种应用至关重要；然而，细胞内传递和转染仍然是困难的任务。虽然在使用化学或物理方法转染实验室建立和维持的（“容易”）细胞系的基础研究应用中，效率高于90%是常见的，但对于“困难”细胞，效率可能降至10%或更低。物理方法的改进（例如，电穿孔）已经实现了性能的逐步改进；然而，目前市场上还没有系统能够满足终端用户对效率、可行性、功能和成本的所有要求。这种新的转染方法是STTR一期项目的主题，有望通过同时创新地使用多种基因转移技术来提高治疗效果，同时通过为困难细胞提供成本效益高的转染解决方案，更好地满足最终用户的需求。

This Small Business Technology Transfer (STTR) Phase I project will prove the technical and commercial feasibility of a single-use polymer microarray for electromechanical transfection of biological cells. Alternative gene transfer solutions are needed to access the potential held in biopharmaceuticals, gene therapy, and stem cell research. Although physical methods for gene transfer (e.g., electroporation) have demonstrated improved (over conventional chemical-mediated techniques) treatment outcomes in difficult-to-transfect primary and stem cells, control and uniformity of treatment remain inadequate. Further, incremental improvements in performance of cuvette-based electroporation systems have only been realized through corresponding increases in system complexity and cost. One-by-one ejection of cells through cell-sized orifices has been found to promote cell membrane poration and DNA delivery into cells by imposing an identical and carefully controlled electromechanical environment on each individual cell. Unfortunately, a relatively high material cost and limitations on achievable treatment conditions (due to constraints on the geometry of microarrays manufactured using standard silicon micromachining techniques), inhibit practical implementation of existing silicon-based microarrays. The innovative design and optimization of the polymer microarray under this STTR Phase I project will yield a low-cost system capable of generating the mechanical stress field needed to achieve improved treatment outcomes in difficult-to-transfect cells.The broader impact/commercial potential of this project is to enable effective and economical transfection of difficult-to-transfect primary and stem cells used for a variety of research and therapeutic applications in the Life Sciences. Effective delivery of genes, drug molecules, imaging agents, peptides, antibodies, and enzymes into living cells is critical to applications ranging from the treatment of human disease through introduction of DNA to the investigation of basic cellular function through single molecule imaging; yet, intracellular delivery and transfection remain difficult tasks. While efficiencies of greater than 90% are common in basic research applications that use chemical or physical methods to transfect laboratory established and maintained ("easy") cell lines, efficiency can drop to 10% or lower for "difficult" cells. Refinements of physical methods (e.g., electroporation) have achieved incremental performance improvements; however, no system currently on the market meets all end-user requirements for efficiency, viability, functionality, and cost. The novel approach to transfection, which is the subject of this STTR Phase I project, promises to improve treatment efficacy through innovative use of multiple gene transfer techniques simultaneously, while better addressing end user needs by providing a cost-effective transfection solution for difficult cells.