Abstract 490P
Background
Colorectal cancer (CRC) patients derived organoids (PDOs) are an increasingly used model for precision medicine. Besides somatic mutations, CRC is characterized by certain levels of copy number alterations (CNAs), which can include oncogenes or tumor suppressors. Moreover, PDOs could be a platform for drug screening.
Methods
Metastatic CRC samples have been obtained from biopsies or surgical samples. After digestion, free cells have been seeded in basement membrane matrix with proper medium. Development of organoids has been observed. After reaching an appropriate volume, organoids have been fixed, stained for hematoxylin and eosin (H&E) and immunohistochemistry (IHC) with common CRC markers. Next generation sequencing (NGS) has been performed with a customized panel. For CNA analysis Cytoscan-HD (Thermo-Scientific) array and data analysis with Chromosome Analysis Suite (Applied Biosystems v4.0) has been performed. For drug assays, PDOs have been plated as single cells and after formation of organoids, chemotherapeutics and biological drugs according to mutational profile have been added. After 96 hours cell viability has been measured with CellTiter-Glo® 3D Reagent.
Results
The success rate in CRC-PDOs establishment is around 80%. PDOs morphology with H&E is comparable to the original tissue and IHC CDX2, CK20 and MUC2/5 positivity. Spectrum of PDOs mutations and polimorphisms is concordant with that of matched patients (R: 0.9). Cytoscan-HD reveals that PDOs maintain mosaicism indicating certain level of heterogeneity. Moreover, PDOs are enriched with CNAs segments. We were able to detect alterations in regions containing genes of interest, such as losses of SMAD2/4, MAP2K4, POLE2 and gains of MYC, CDX2, CARD11, AURKA and many others. Drug treatment showed a good concordance with patient response. Furthermore, we were able to confirm synergy of chemotherapeutics combination in our PDO’s models.
Conclusions
Metastatic CRC-PDOs recapitulate morphological and genomic features of the original patient. In addition to mutational profile, CNAs pattern allows to better capture genomic alterations. Drug assays are feasible and could be a valuable tool to predict patient’s response.
Clinical trial identification
Editorial acknowledgement
Legal entity responsible for the study
The authors.
Funding
This study was supported by grants from the Carlos III Health Institute (PI18/01508 to TF; PI15/02180 and PI18/01909 to AC). MC is supported by a pre-doctoral grant from the Spanish Cancer Association (AECC), Spain. FP is a recipient of an ESMO translational research grant. MFGB is a recipient of a Grisolia predoctoral grant (GRISOLIAP/2017/161) by the Conselleria de Educación, Investigación, Cultura y Deporte of Valencia. VG is supported by a Rio Hortega contract CM18/00241 from the Carlos III Health Institute. NT was supported by Rio Hortega contract CM15/00246 from the Carlos III Health Institute. DR was supported by Joan Rodes contract 16/00040 from the Carlos III Health Institute. TF is supported by Joan Rodes contract 17/00026 from the Carlos III Health Institute.
Disclosure
A. Cervantes: Advisory/Consultancy, Speaker Bureau/Expert testimony, Research grant/Funding (self), Research grant/Funding (institution): Merck Serono; Advisory/Consultancy, Speaker Bureau/Expert testimony, Research grant/Funding (self), Research grant/Funding (institution): Roche; Advisory/Consultancy, Research grant/Funding (self), Research grant/Funding (institution): Beigene; Advisory/Consultancy, Speaker Bureau/Expert testimony, Research grant/Funding (self), Research grant/Funding (institution): Bayer; Advisory/Consultancy, Speaker Bureau/Expert testimony, Research grant/Funding (self), Research grant/Funding (institution): Servier; Advisory/Consultancy, Research grant/Funding (self), Research grant/Funding (institution): Lilly; Advisory/Consultancy, Research grant/Funding (self), Research grant/Funding (institution): Novartis; Advisory/Consultancy: Takeda; Advisory/Consultancy, Research grant/Funding (self), Research grant/Funding (institution): Astelas; Advisory/Consultancy: Pierre Fabre; Research grant/Funding (self), Research grant/Funding (institution): Genentech; Research grant/Funding (self), Research grant/Funding (institution): Fibrogen; Research grant/Funding (self), Research grant/Funding (institution): Amcure; Research grant/Funding (self), Research grant/Funding (institution): Sierra Oncology; Research grant/Funding (self), Research grant/Funding (institution): Astra Zeneca; Research grant/Funding (self), Research grant/Funding (institution): Medimmune; Research grant/Funding (self), Research grant/Funding (institution): BMS; Research grant/Funding (self), Research grant/Funding (institution): MSD; Speaker Bureau/Expert testimony: Amgen; Speaker Bureau/Expert testimony: Foundation Medicine. All other authors have declared no conflicts of interest.