Surefection Technology for High-Throughput Primary Cell Transfection

描述（由申请人提供）：人原代细胞是直接从患者或健康供体的组织样本中提取的细胞。它们是人类生理和疾病的最真实的体外模型，因此在医学发现和发展中是无价的。使用人类原代细胞作为生理或疾病相关模型，为更好地了解和治疗各种人类疾病（包括癌症、心血管疾病和炎症性疾病）的研究和开发提供了显著优势。不幸的是，由于缺乏副作用最小的有效转染手段，特别是在高通量模式下，宝贵原代细胞的利用在很大程度上受到阻碍。为了解决这一未满足的需求，我们提出开发一种SureFection技术，该技术结合了细胞转染领域的两个新兴进展，即受控原位电穿孔和DMA核递送，以有效转染原代细胞而无生化副作用。该策略将在新型微型设备中实现，该设备可以很容易地复用，以符合96孔载玻片/1536孔微孔板uHT足迹，允许大规模的体内基因功能分析，以及使用原代细胞进行药物筛选和测试。在第一阶段，我们将展示我们的技术在三种原代细胞系统中的适用性，这些原代细胞系统是研究乳腺癌和前列腺癌、血管生成和心血管疾病的重要模型。我们的长期目标是将这项技术与我们新颖的功能分析方法相结合，为目标识别和验证、化合物筛选和体外药物测试/分析提供基于原代细胞的高通量解决方案，作为我们促进更快、更便宜和更安全的小分子药物开发战略的一部分。拟议的研究和开发将促进我们对人类基因在生理和疾病中的作用的理解，并加速治疗癌症、心血管和其他疾病的开发，从而有利于公共卫生。

DESCRIPTION (provided by applicant): Human primary cells are cells directly derived from tissue samples from patients or healthy donors. They are the truest in vitro models of human physiology and disease, therefore are invaluable in medical discovery and development. Use of human primary cells as physiology- or disease-relevant models provides significant advantages in research and development for better understanding and treatment of various human diseases, including cancer, cardiovascular and inflammatory diseases. Unfortunately, utility of precious primary cells is largely hindered by the lack of means for effective transfection with minimal side-effects, particularly in high- throughput mode. To address this unmet need, we propose the development of a SureFection technology that combines two emerging advancements in the field of cell transfection, i.e., controlled in situ electroporation and DMA nuclear delivery, to effective transfect primary cells free of biochemical side-effects. This strategy will be implemented in novel miniature devices that can be readily multiplexed to conform to 96-well slide/1536-well microplate uHT footprints, permitting massive scale in vivo gene funtion analysis, as well as drug screening and testing using primary cells. In Phase I, we will demonstrate the applicability of our technology in three primary cell systems that are important models for study of breast and prostate cancer, angiogenesis and cardiovascular disease. Our long-term goal is to combine this enabling technology with our novel functional assay methods to provide primary cell based high-throughput solutions for target ID&validation, compound screening and in vitro drug testing/profiling, as part of our strategies to facilitate faster, cheaper and safer small molecule drug development. The proposed research and development will benefit public health by forwarding our understanding of the roles of human genes in physiology and disease, and accelerating therapeutic development to fight with cancer, cardiovascular and other diseases.