A Novel High Throughput Tumor Spheroid Microtechnology

一种新型高通量肿瘤球体显微技术

基本信息

批准号：
8738627
负责人：
Gary D Luker
金额：
$ 24.57万
依托单位：
UNIVERSITY OF AKRON
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-20 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8738627
关键词：
Academia Address Adherent Culture Biological Assay Breast Cancer Cell Cancer Biology Cell Aggregation Cell Culture System Cell Density Cell Survival Cells Cellular Spheroids Clinical Complex Consumption Culture Media Development Dextrans Diffusion Drops Drug Compounding Drug Formulations Drug Targeting Ensure Environment Equipment Exhibits Gene Expression Generations Goals Growth Image Immersion Investigative Technique Industry Kinetics Laboratories Liquid substance Mainstreaming Maintenance Malignant Neoplasms Malignant Stromal Cell Metabolic Methods Modeling Molecular Molecular Profiling Morphology Nutrient Osmolalities Oxygen Pharmaceutical Preparations Phase Physiological Physiology Polyethylene Glycols Polymers Preclinical Drug Evaluation Process Reader Reagent Research Personnel Robotics Solid Neoplasm Stromal Cells Stromal Neoplasm Surface System Techniques Technology Testing Time Validation anticancer research aqueous cancer cell cancer therapy chemotherapy clinical efficacy cost effective cost effectiveness dextran drug development drug discovery drug testing evaporation high standard high throughput screening improved in vivo instrument nanolitre neoplastic cell novel physiologic model public health relevance response screening technology validation tumor tumor microenvironment two-dimensional

项目摘要

DESCRIPTION (provided by applicant): Two-dimensional (2D) cultures of cancer cells are routinely used in drug discovery for screening and initial characterization of the efficacy of librry of potential drug compounds. Despite their simplicity and compatibility with high throughput screening instruments, 2D cell assays often fail to predict the efficacy of compounds in vivo, making drug development and discovery an extremely costly process. Disparity between 2D cultures and the complex 3D environment of cancer cells in vivo is the major shortcoming of monolayer culture systems. Development of novel chemotherapeutics requires compound screening against malignant tumor cells in a setting that resembles the 3D tumor environment. Cancer cell spheroids (CCS) are 3D clusters of malignant cells that are regarded as physiologic models of solid tumors; they possess similar metabolic and proliferative gradients to avascular tumors and exhibit the clinical expression profiles of solid tumors. Despite the inherent power of CCS to predict clinical efficacy of drugs, incorporation of CCS into the mainstream drug development process is severely hindered by complex and expensive methodological requirements for the formation and maintenance of CCS. We overcome the limitations of existing techniques by developing a technological platform to generate spheroids of consistent size in standard 384-microwell plates using an aqueous two- phase system (ATPS). A drop of the denser aqueous phase containing cancer cells is robotically dispensed into each well containing the second, immersion phase. The drop confines cells and remains immiscible from the immersion phase to facilitate aggregation of cells into a compact CCS of well-defined size. Importantly the overlay of culture media provides nutrients and minimizes the well-known problem of evaporation and changes in osmolality of media as in other assays. The entire process of generating 384 spheroids is done robotically and in a single step. The unprecedented ease of formation and maintenance of CCS and full compatibility with standard equipment in high throughput screening laboratories makes this microtechnology readily available to the researchers in academia and industry. We anticipate that this microtechnology will make drug testing and screening with 3D tumor models a routine laboratory technique prior to expensive and tedious in vivo analyses. In addition, it will dramatically improve testing throughput and cost-effectiveness (increasing numbers of tested compounds and reduced reagent consumption) and efficiency (reducing hands-on time) to expedite drug discovery. We will accomplish our goals through two specific aims: (i) Generation of cancer cell spheroids with aqueous biphasic systems; (ii) Initial validation of ATPS spheroids for compound testing.

描述（由申请人提供）：癌细胞的二维（2D）培养物通常用于药物发现中，用于筛查和初始表征潜在药物化合物的库的功效。尽管它们与高吞吐量筛选工具的简单性和兼容性，但2D细胞测定法通常无法预测体内化合物的功效，从而使药物开发和发现成为极为昂贵的过程。 2D培养物与体内癌细胞复杂的3D环境之间的差异是单层培养系统的主要缺点。在类似于3D肿瘤环境的环境中，新型化学治疗剂的开发需要针对恶性肿瘤细胞进行复合筛查。癌细胞球体（CC）是恶性细胞的3D簇，被视为实体瘤的生理模型。它们具有与血管肿瘤相似的代谢和增殖梯度，并表现出实体瘤的临床表达谱。尽管CC可以预测药物的临床功效的固有力量，但将CCS纳入主流药物开发过程中，对CC的形成和维持的复杂和昂贵的方法论要求严重阻碍了CC。我们通过开发技术平台来克服现有技术的局限性，以使用水性两相系统（ATP）在标准384-microwell板上生成一致尺寸的球体。将含有癌细胞的浓密水相滴入机器人中，并将其分配到包含第二个浸入阶段的每个井中。滴剂将细胞限制在浸入阶段，以促进细胞聚集到定义明确的尺寸的紧凑型CC中。重要的是，培养基的覆盖层提供了营养，并最大程度地减少了媒体渗透性的蒸发和渗透变化的问题，就像其他测定中一样。生成384个球体的整个过程是通过机器人和一个步骤完成的。 CCS的形成和维护的前所未有的易度性以及与高通量筛选实验室中的标准设备的完全兼容性，使该微观技术很容易为学术界和行业的研究人员提供。我们预计，这种微技术将在昂贵且乏味的体内分析之前，通过3D肿瘤模型将药物测试和筛查成为常规的实验室技术。此外，它将显着改善测试吞吐量和成本效益（测试化合物数量的增加和减少的试剂消耗量）和效率（减少动手时间）以加快药物发现。我们将通过两个具体的目标来实现我们的目标：（i）用两类水性系统产生癌细胞球体；（ii）对ATP的初始验证用于复合测试。