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.

Public Health Relevance

Effective targeting of malignant tumor cells is fundamental to successful treatment of cancers. Unfortunately existing chemotherapy drugs do not effectively eliminate tumor cells, in large due to the disparity between the three-dimensional (3D) tumor environment in vivo and two-dimensional (2D) cultures used for drug testing. We will develop a new technological platform to generate highly consistent 3D tumor models and enable simultaneous screening of hundreds of drug compounds in an efficient and cost-effective manner. This technology will streamline drug discovery and significantly expedite identification of novel and more potent drugs against various cancers.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21CA182333-02
Application #
8738627
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Sorg, Brian S
Project Start
2013-09-20
Project End
2015-08-31
Budget Start
2014-09-01
Budget End
2015-08-31
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Akron
Department
Biomedical Engineering
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
City
Akron
State
OH
Country
United States
Zip Code
44325
Ham, Stephanie L; Joshi, Ramila; Luker, Gary D et al. (2016) Engineered Breast Cancer Cell Spheroids Reproduce Biologic Properties of Solid Tumors. Adv Healthc Mater 5:2788-2798
Ham, Stephanie L; Joshi, Ramila; Thakuri, Pradip S et al. (2016) Liquid-based three-dimensional tumor models for cancer research and drug discovery. Exp Biol Med (Maywood) 241:939-54
Stacer, A C; Fenner, J; Cavnar, S P et al. (2016) Endothelial CXCR7 regulates breast cancer metastasis. Oncogene 35:1716-24
Ham, Stephanie L; Thakuri, Pradip S; Tavana, Hossein (2015) Robotic printing and drug testing of 384-well tumor spheroids. Conf Proc IEEE Eng Med Biol Soc 2015:2183-6
Atefi, Ehsan; Joshi, Ramila; Mann Jr, Jay Adin et al. (2015) Interfacial Tension Effect on Cell Partition in Aqueous Two-Phase Systems. ACS Appl Mater Interfaces 7:21305-14
Ham, Stephanie Lemmo; Nasrollahi, Samila; Shah, Kush N et al. (2015) Phytochemicals potently inhibit migration of metastatic breast cancer cells. Integr Biol (Camb) 7:792-800
Chang, S Laura; Cavnar, Stephen P; Takayama, Shuichi et al. (2015) Cell, isoform, and environment factors shape gradients and modulate chemotaxis. PLoS One 10:e0123450
Stacer, Amanda C; Wang, Hanxiao; Fenner, Joseph et al. (2015) Imaging Reporters for Proteasome Activity Identify Tumor- and Metastasis-Initiating Cells. Mol Imaging 14:414-28
Xiang, Jingyu; Hurchla, Michelle A; Fontana, Francesca et al. (2015) CXCR4 Protein Epitope Mimetic Antagonist POL5551 Disrupts Metastasis and Enhances Chemotherapy Effect in Triple-Negative Breast Cancer. Mol Cancer Ther 14:2473-85
Ham, Stephanie Lemmo; Atefi, Ehsan; Fyffe, Darcy et al. (2015) Robotic production of cancer cell spheroids with an aqueous two-phase system for drug testing. J Vis Exp :e52754

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