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：
8625056
负责人：
Gary D Luker
金额：
$ 25.75万
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 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8625056
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，以筛选和初步表征潜在药物化合物库的功效。尽管二维细胞检测简单且与高通量筛选仪器兼容，但它们通常无法预测化合物在体内的功效，使得药物开发和发现成为一个极其昂贵的过程。 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。新型化疗药物的开发需要在类似于 3D 肿瘤环境的环境中针对恶性肿瘤细胞进行化合物筛选。癌细胞球体 (CCS) 是恶性细胞的 3D 簇，被视为实体瘤的生理模型；它们具有与无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CCS具有预测药物临床疗效的固有能力，但将CCS纳入主流药物开发过程却受到CCS形成和维护复杂且昂贵的方法学要求的严重阻碍。我们通过开发一个技术平台来克服现有技术的局限性，使用水性两相系统 (ATPS) 在标准 384 微孔板中生成尺寸一致的球体。将一滴含有癌细胞的较致密水相自动分配到含有第二浸入相的每个孔中。该液滴限制了细胞并在浸入阶段保持不混溶，以促进细胞聚集成尺寸明确的紧凑CCS。重要的是，培养基的覆盖提供了营养，并最大限度地减少了众所周知的蒸发问题和培养基渗透压的变化，如其他测定中一样。生成 384 个球体的整个过程都是由机器人一步完成的。 CCS 的形成和维护前所未有的简便性以及与高通量筛选实验室标准设备的完全兼容性使得学术界和工业界的研究人员可以轻松使用这种微技术。我们预计，这种微技术将使 3D 肿瘤模型的药物测试和筛选成为常规实验室技术，取代昂贵且繁琐的体内分析。此外，它将显着提高测试吞吐量和成本效益（增加测试化合物的数量并减少试剂消耗）和效率（减少实践时间），以加快药物发现。我们将通过两个具体目标来实现我们的目标：（i）利用水性双相系统生成癌细胞球体； (ii) 用于化合物测试的 ATPS 球体的初步验证。