Advanced Development of Primary Pancreatic Organoid Tumor Models for High Throughput Phenotypic Drug Screening
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Published abstract. For the performed magnetic 3D assay, standard flat bottom 384 well plates and 1536 well plates featuring a cell repellent surface were employed, combined with the magnetic 3D bioprinting technology. The homogeneous HTS organoid-based assay was standardized using well known anti-cancer agents against four patient-derived pancreatic cancer KRAS mutant associated primary cell lines, including cancer-associated fibroblasts. This technology was further validated for its compatibility with HTS instrumentation and robotics by completing a cytotoxicity screen with a library of 3300 approved drugs. To determine the validity of 3D vs 2D HTS, a parallel screen was performed on the same cells in monolayer culture. These results indicate that an ex-vivo clinically relevant 3D tumor model can be rapidly adapted and used for large scale drug screening, taking us one step closer to applying precision medicine to the treatment of cancer.
Study Conclusions
Validated 3D cell culture using magnetic 3D bioprinting (M3D) technology for HTS drug screening against solid tumor primary cells. Most of the tested compounds showed less activity in 3D. However, there were drugs that showed preferential cytotoxicity against cells cultured in 3D over 2D. Notably, disulfiram was active in 3D models only, and it has been reported in initial clinical studies its efficacy in metastatic pancreatic cancer, prostate cancer, and glioblastoma. These results underscore the phenotypical screening relevance of culturing cells in 3D with M3D. The reported findings showed that this approach is robust and rapid.
Future direction
- Efforts will be directed on the screening of larger chemical libraries (~150,000) in a 1536-well automated platform
- Explore the biology of co-culturing pancreatic cancer cells and its cancer-associated fibroblast (CAF) as an improved model to predict the response of drugs to the treatment of patients with pancreatic cancer
- Follow up studies in mouse models of pancreatic tumors
Cell types listed
- Human colorectal adenocarcinoma cell line HT-29 (ATCC no. HTB-38) and human pancreatic epithelial carcinoma cell line PANC-1 (ATCC no. CRL-1469)
- Primary human pancreatic ductal cells hM1 and hT1 (immortalized by SV40), were generated from tissues of pancreatic cancer patients
- Primary cancer-associated fibroblasts hM1-CAF and hT1-CAF (immortalized by SV40), were generated from tissues of pancreatic cancer patients
M3D system used
- 384-well bioprinting
- 1,536-well bioprinting
Topics
- HTS
- Pancreatic cancer
- Organoids
- 3D cell culture
- 3D magnetic bioprinting
“It is exciting that we picked the ‘needle in a haystack. We identified, out of the 3,300 approved drugs, the compound that is currently undergoing Phase I clinical trials for the treatment of pancreatic cancer, disulfiram. Disulfiram was only active in certain cancer specific 3D cultures, not in 2D.”
Key findings
In this work, magnetic 3D bioprinting provided key strategic advantages, since it is a rapid, relatively easy, and reproducible method to print 3D cultures in high throughput, including these key points:
- validated 384- and 1536-well magnetic 3D bioprinting systems as a powerful tool for HTS
- homogeneous HTS organoid-based assay
- standardized using well known anti-cancer agents against four patient-derived pancreatic cancer KRAS mutant associated primary cell lines
- screen included cancer-associated fibroblasts (CAF)
- validated for its compatibility with HTS instrumentation and robotics
- cytotoxicity screen with a library of 3300 FDA approved drugs
- ex-vivo clinically relevant 3D tumor model can be rapidly adapted and used for large scale drug screening, taking us one step closer to applying precision medicine to the treatment of cancer
- compared 3D vs. 2D HTS of 3,300 compound FDA library (FDA approved compounds)
- 2D showed many more active compounds which should not, "false positives” result
- 3D identified compound that is currently undergoing clinical trial for pancreatic cancer, 2D did not
- HTS done with flat-bottom plates in 384- and 1536-well
“This work demonstrates a faster and more physiologically relevant cost-efficient drug discovery process, which ultimately will aid in avoiding possible false positives and improving the accuracy.”
Academic Authors, key opinion leaders (KOL)
- Scripps Research Institute
- University of Texas
- Cold Spring Harbor Laboratory
- Danna Farber Cancer Institute
- Full Reference: SLAS Discov. 2018 Jul;23(6):574-584. doi: 10.1177/2472555218766842. Epub 2018 Apr 19.
- Original Journal: SLAS Discovery
