İlaç yanıtının tahmini için hasta kaynaklı tümör organoidleri

A key limitation in cancer treatment is the lack of valid predictive biomarkers, which reduces the efficacy of treatments. Personalized medicine strategies include protein-, RNA-based and genome based stratification, though in oncology, precision medicine has been largely based on genomic biomarkers. However, less than half of patients are eligible for genetically matched treatment and for the majority of anticancer agents no genetic markers are available. A promising predictive biomarker is individualized tumor response testing using patient-derived organoids (PDOs), in which anticancer agents are screened ex vivo on PDOs to predict clinical response. PDOs have been developed for a variety of tumors and are stem-cell derived, three-dimensional self- organizing structures comprised of epithelial cells, mimicking its corresponding tumor. This study aims to establish the organoids of colorectal cancer (CRC) patients by using in-house protocol and to predict drug response. The patient cohort of our study consists of CRC patients over the age of 18. Fresh surgical material taken from the patients reached our laboratory within 24 hours at +4oC. After dissociating the tumor tissue mechanically and chemically, cells were counted and embedded in matrigel. After 3-4 week culture, organoids were tyrpsinized into single cell and passaged into matrigel containing 96 well plate as 6000 cells/5ul dome. Whenever organoids reformed, drug (5-FU, FOLFOX, FOLFIRI, CAPOX,..) were added in different concentrations. After 7 day incubation, viability analyses were performed by CellTiter-Glo 3D Viability Assay kit. All experiments were replicated four times and statistical analyses were done by one-way ANOVA. Finally, IC50 of drugs were calculated by GraphPad Prism 8 program. We were able to establish organoids from tumor tissues with a success rate of 90%. By performing drug screening for various anticancer agents on these organoids, we determined that each patient's response to treatments is different. We have seen that targeted antibodies increase the efficacy of treatment in one patient. We have successfully predict drug responses for 5 patients as a proof of concept study.