Optimizing pancreatic cancer organoids for personalized medicine

• Applicant: Dr. Benno Traub
• Subject Area: General and Visceral Surgery ; Gastroenterology
• Term: from 2018 to 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 419258639

Pancreatic cancer is known for its high mortality rate which is equal to its incidence. It is characterized by a rapid progression of the disease with a mean progression free survival of 4 months and a mean overall survival of 7 months. In order to develop new therapeutic approaches, an adequate cancer model is a prerequisite.Prof. Tuveson and his team at Cold Spring Harbor Laboratories were able to develop an organoid model of human pancreatic carcinoma. These organoids can be generated from very few originating cells and are characterized by high genetic and morphological stability compared to the original tumor. This allows the use of tumor organoids in the field of personalized medicine. Pancreatic cancer organoids have been shown to be a predictive model for patient response. This approach can be offered complimentary for testing the most effective tumor-specific therapy.As recently shown by the group of Prof. Tuveson, organoid cultures currently require at least 4-6 weeks for establishing with a generation efficiency of 75% to achieve a sufficient size for in vitro studies. However, in order to be able to use the strengths of this model in a clinically meaningful manner, the generation speed and efficiency must be further optimized.For this purpose, in the current research project established organoid cultures are used as so-called "feeder cells/feeder organoids". The feeder cells grow in the same culture as the target culture of a new tumor specimen. Using feeder cells we assume that organoid generation can be accelerated by secretion of growth factors and extracellular matrix proteins as well as cell to cell interactions. Through empirical testing of different cell lines such as feeder organoids and non-malignant, so-called cancer-associated fibroblasts, the best culture condition will be evaluated.To enable the chemotherapy treatment of the organoids later on, the established co-culture needs to be cleared from the feeder cells. The established feeder cell line will be transfected by using a CRISPR-knock-in with a suicide gene that is fused to a fluorescent protein.Based on the further characterization of the identified feeder cell line by genome and secretome analysis, the growth requirements of human pancreatic carcinoma will be further investigated.Ultimately, the new organoid model will undergo drug testing. By comparison with existing data of the Tuveson laboratory and comparing derived models from different feeder cells, an influence of the co-culture on the treatment sensitivity should be excluded.The current research project serves the further investigation of growth requirements and interactions with surrounding cells of pancreatic adenocarcinoma. By using the organoid model, this is done in the closest possible proximity to clinical utility. The optimized organoid model can thus enable the fastest possible patient treatment in personalized medicine with effective, specific therapies.

• DFG Programme: Research Fellowships
• International Connection: USA
• Host: Professor David A. Tuveson, Ph.D.