Electrosonic Ejector Microarray for Development of Cancer Therapies

描述(申请人提供)：尽管几十年来的研究主要集中在基因转染上，但有效的细胞内输送生物相关材料仍然是一项艰巨的任务，阻碍了各种生物和生物医学领域的研究。这项拟议的工作描述了一种商业上可行的细胞内纳米材料递送装置，电声喷射器微阵列(EEM)，它通过精确控制生物物理作用(即同时应用声/机械穿孔、电穿孔和热复制)来优化药物和/或基因/核酸和/或显像剂进入细胞的递送。EEM是一种新型的微电子机械系统(MEMS)设备，它通过带有集成的电穿孔电极的微型喷嘴喷射包含生物细胞的样本，从而通过机械/电穿孔相结合的方式在细胞膜上开孔以摄取纳米材料。1高频操作和并行(阵列)格式使能够以每秒1到1亿个细胞的速度快速处理大细胞群体；然而，当以连续流动模式操作时，该设备可能可以容纳从~100 NL到任意大容量的广泛的样本大小。此外，该设备可以一次性使用，以消除交叉污染，并在单细胞水平上提供统一的(即，在整个细胞群体中相同的)处理，这是在细胞生物学和基因治疗的临床应用中制备样品的关键能力。EEM非常适合基础/应用研究以及诊断和治疗用途。初步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。通过这项拟议的工作将获得的对EEM操作参数和实现的生物效应之间关系的更好的理解可以直接转移到其他应用领域。这项拟议工作的主要目标是开发一种商业上可行的EEM，证明比目前可用的纳米材料输送技术有定量的性能改进。为了实现这一目标，(1)将开发一个针对细胞治疗进行优化的EEM平台，(2)将展示各种细胞类型的安全治疗和纳米材料定位的特征，(3)将优化EEM的转染能力，并量化其相对于传统转基因技术的优势。公共卫生相关性：电声喷射器微阵列(EEM)的开发将解决当前细胞内药物/基因传递技术的不足，这些技术阻碍了各种生物和生物医学领域的研究。特别是，成功地将EEM用于治疗胶质母细胞瘤的治疗方法的开发，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.