Electrosonic Ejector Microarray for Development of Cancer Therapies

描述（由申请人提供）：尽管几十年来的研究主要集中在基因转染上，但有效的细胞内递送生物学相关材料仍然是一项艰巨的任务，抑制了各种生物学和生物医学领域的研究。这项工作描述了一种商业上可行的细胞内纳米材料递送装置，即电子弹射微阵列（EEM），它通过精确控制生物物理作用（即同时应用声/机械穿孔、电穿孔和热穿孔），优化了药物和/或基因/核酸和/或显像剂递送到细胞中。EEM是一种新型的微机电系统（MEMS）设备，它通过带有电穿孔电极的微观喷嘴喷射含有生物细胞的样品，从而通过机械/电穿孔结合在细胞膜上打开孔隙，以吸收纳米材料操作的高超声波频率和并行（阵列）格式能够以每秒1至1亿个细胞之间的速率快速处理大细胞群；然而，当在连续流模式下操作时，该设备可以容纳大范围的样品尺寸，从~100 nL到任意大体积。此外，该设备可以一次性制造，以消除交叉污染，并在单细胞水平上提供均匀（即在整个细胞群中相同）的处理，这是细胞生物学和基因治疗临床应用中样品制备的关键能力。EEM非常适合基础/应用研究，以及诊断和治疗用途。最初的研究将集中在将成熟的重组蛋白疗法与新兴的RNA干扰（RNAi）技术相结合的癌症治疗上。通过这项提议的工作将获得的EEM操作参数和实现的生物效应之间关系的改进理解可直接转移到其他应用领域。这项工作的主要目标是开发一种商业上可行的EEM，以证明比目前可用的纳米材料传输技术的定量性能改进。为了实现这一目标，(1)将开发一个优化细胞处理的EEM平台，(2)将展示各种细胞类型的安全处理和纳米材料定位的表征，(3)将优化EEM的转染能力，并量化其相对于传统转染技术的优势。公共卫生相关性：电子弹射微阵列（EEM）的发展将解决当前细胞内药物/基因递送技术的不足，这些技术正在抑制各种生物和生物医学领域的研究。特别是，EEM在治疗胶质母细胞瘤（最常见和致命的脑肿瘤）的疗法开发中的成功应用，将证明其在广泛的药物发现和基因治疗应用中的可行性。

DESCRIPTION (provided by applicant): Despite decades of research focused mainly on gene transfection, effective intracellular delivery of biologically relevant material remains a difficult task, inhibiting research in a variety of biological and biomedical fields. This proposed work describes a commercially viable intracellular nanomaterial delivery device, the Electrosonic Ejector Microarray (EEM), that is optimized for drug and / or gene / nucleic acid and / or imaging agent delivery into cells via precise control of biophysical action (i.e., concurrent application of sono / mechanoporation, electroporation and thermoporation). The EEM is a novel microelectromechanical systems (MEMS)-enabled device that ejects a sample containing biological cells through microscopic nozzles with incorporated electroporation electrodes, thereby opening pores in the cell membrane via combined mechano / electroporation for uptake of nanomaterials.1 The high ultrasonic frequency of operation and a parallel (array) format enable fast processing of large cell populations at rates between 1 and 100 million cells per second; however, the device can potentially accommodate a wide range of sample sizes, from ~100 nL to arbitrarily large volumes when operated in continuous-flow mode. In addition, the device can be made disposable to eliminate cross-contamination and provides uniform (i.e., identical across an entire cell population) treatment on a single-cell level, which are critical capabilities for sample preparation in clinical applications of cell biology and gene therapy. The EEM is well-suited to basic / applied research, as well as diagnostic and therapeutic uses. Initial investigations will focus on cancer therapies combining mature recombinant protein therapies with emerging RNA interference (RNAi) technologies. The improved understanding of the relationship between the EEM operating parameters and realized bioeffects that will be gained through this proposed work is directly transferable to other application spaces. The primary objective of the proposed work is to develop a commercially viable EEM that demonstrates quantitative performance improvement over currently available nanomaterial delivery technologies. To achieve this objective, (1) an EEM platform that is optimized for cell treatment will be developed, (2) safe treatment of a variety of cell types and characterization of nanomaterial localization will be demonstrated, and (3) the transfection capabilities of the EEM will be optimized and its advantages over traditional transfection technologies quantified. PUBLIC HEALTH RELEVANCE: Development of the Electrosonic Ejector Microarray (EEM) will address the deficiencies of current intracellular drug / gene delivery techniques, which are inhibiting research in a variety of biological and biomedical fields. In particular, successful use of the EEM in development of therapies to treat glioblastomas, which are the most common and lethal brain tumors, will prove its feasibility in a wide range of drug discovery and gene therapy applications.