Two-dimensional (2D) tissue culture models are highly simplified cancer models unable to capture the complexity and heterogeneity found in-vivo. Around 95% of new anticancer drugs eventually fail in clinical trial despite robust indications of activity in existing in vitro pre-clinical models, making in vitro testing some of the least predictive. Three dimensional (3D) spheroid culture models have recently advanced to bridge the ?in- vitro to in-vivo gap? and provide the means for assembling more complex cancer relevant tissue microenvironments. Although these 3D models are being adopted by industry and the academic community, they have limitations and are hampered by low throughput, lack of consistency, high costs and the need for clinical validation. The Scripps Research Institute Molecular Screening Center (SRIMSC) in partnership with n3D Biosciences Inc., Greiner Bio-One USA Inc., Dr. Derek Duckett at Scripps Research department of Molecular Therapeutics and Dr. David Tuveson, M.D, Ph.D. at Cold Spring Harbor Laboratory (CSHL), have created a strategic collaboration to advance a novel technology known as 3D magnetic bioprinting. Magnetic 3D bioprinting addresses the these critical issues by utilizing n3D's core technology known as the NanoShuttle to levitate and aggregate cells using magnetic forces to produce spheroids/organoids. The ultimate end product will be an affordable; HTS validated 384 and 1536 microplate format that supports rapid/consistent production of 3D spheroids for a wide array of cell types including primary tumor lines. The end goal is to accelerate 3D spheroid cultivation using screening automation, improve cost efficiency and allow for rapid drug testing such as FDA approved drugs in reformulation/repurposing studies. Advancement of this technology will be facilitated through the following:
Aim 1 : Validation of the current 384 well plate nanosphere technology in a HTS facility for automation compatibility. Compare 3D results to 2D models of KRAS pancreatic cancer cell models as provided by Dr. Tuveson.
Aim 2 : Validation of n3D spheroid technology for drug testing against select cytotoxic drugs, NCI approved oncology drug set and the Scripps FDA Approved drug collection. CC50 data, i.e. the concentration that produces 50% cellular cytotoxicity, in 2D and in 3D formats will be compared to published literature.
Aim 3 : n3D Biosciences will produce an advance 1536 well plate NanoShuttle driver compatible for HTS and drug discovery efforts. SRIMSC will evaluate and implement the higher density format for drug discovery utility which will culminate in its testing on a large library of ~150K compounds to demonstrate HTS readiness.
Aim 4 : The n3D spheroid technology will be employed against patient derived primary Glioblastoma Multiform (GBM) derived cells with the end goal of evaluating its utility in primary cancer cell research.
Aim 5 : The n3D spheroid technology will be evaluated in-vivo for pancreatic orthotopic tumor effect and its utility in preclinical research. The end goal is to transfer and implement this technology and methods worldwide for cancer research and early drug discovery.
The goal of this project is to advance magnetic bioprinting to produce 3D tissue culture relevant models that are more representative of the complexity found in cancer providing greater accuracy in evaluating anti-cancer agents. This will be made available worldwide to the biomedical research community focused on cancer biology research, early drug discovery and clinical studies. Success will improve the quality and speed of 3D spheroid manufacturing and have a direct impact on; (i) high throughput screening automation providing a cost effective screening assay for drug discovery; (ii) enhance basic cancer research providing a more accurate in vitro model; (iii) accelerate FDA approved drug repurposing and reformulation studies that are needed to establish precision/personalize medicine against cultured biopsies from cancer patients.
|Wolff, Robert A; Wang-Gillam, Andrea; Alvarez, Hector et al. (2018) Dynamic changes during the treatment of pancreatic cancer. Oncotarget 9:14764-14790|
|Kota, Smitha; Hou, Shurong; Guerrant, William et al. (2018) A novel three-dimensional high-throughput screening approach identifies inducers of a mutant KRAS selective lethal phenotype. Oncogene 37:4372-4384|
|Hou, Shurong; Tiriac, Hervé; Sridharan, Banu Priya et al. (2018) Advanced Development of Primary Pancreatic Organoid Tumor Models for High-Throughput Phenotypic Drug Screening. SLAS Discov 23:574-584|
|Singhera, Fakhar; Cooper, Emily; Scampavia, Louis et al. (2018) Using bead injection to model dispensing of 3-D multicellular spheroids into microtiter plates. Talanta 177:74-76|
|Quereda, Victor; Hou, Shurong; Madoux, Franck et al. (2018) A Cytotoxic Three-Dimensional-Spheroid, High-Throughput Assay Using Patient-Derived Glioma Stem Cells. SLAS Discov 23:842-849|
|Hou, Shurong; Madoux, Franck; Scampavia, Louis et al. (2017) Drug Library Screening for the Identification of Ionophores That Correct the Mistrafficking Disorder Associated with Oxalosis Kidney Disease. SLAS Discov 22:887-896|
|Madoux, Franck; Tanner, Allison; Vessels, Michelle et al. (2017) A 1536-Well 3D Viability Assay to Assess the Cytotoxic Effect of Drugs on Spheroids. SLAS Discov 22:516-524|