Surefection Technology for High-Throughput Primary Cell Transfection

描述(申请人提供)：人类原代细胞是直接从患者或健康捐赠者的组织样本中提取的细胞。它们是人类生理和疾病最真实的体外模型，因此在医学发现和发展中具有无价的价值。使用人类原代细胞作为与生理或疾病相关的模型在研究和开发方面提供了显著的优势，有助于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。遗憾的是，由于缺乏有效的、副作用最小的转基因手段，尤其是在高通量模式下，珍贵的原代细胞的实用性在很大程度上受到了阻碍。为了解决这一未得到满足的需求，我们建议开发一种SureFect技术，该技术结合了细胞转染领域的两个新进展，即受控原位电穿孔和DMA核传递，以有效地转染原代细胞，而没有生化副作用。这一战略将在新型微型设备中实施，这些设备可以很容易地进行多路复用，以符合96孔幻灯片/1536孔微板UHT足迹，从而允许进行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个原代细胞系统中的适用性，这三个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一使能技术与我们新的功能分析方法相结合，为目标ID验证、化合物筛选和体外药物测试/图谱提供基于原代细胞的高通量解决方案，作为我们战略的一部分，以促进更快、更便宜和更安全的小分子药物开发。拟议的研究和开发将促进我们对人类基因在生理学和疾病中的作用的理解，并加快抗击癌症、心血管和其他疾病的治疗开发，从而使公众健康受益。

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.