Abstract 2786
Background
Organoids are 3D in vitroprimary culture of great interest for translational research representing an efficient and reproducible cancer model. The aim of this project is to generate a biobank of colorectal cancer (CRC) patients derived organoids (PDOs) that could be used to analyze molecular characteristics and to test different treatments as well as to study the underlying molecular causes of cancer and treatment resistance.
Methods
Primary or metastatic CRC tissues have been obtained from patients who underwent surgery. Tissue has been washed and incubated with antibiotics. After mechanical and enzymatic digestion, free cells have been seeded in Matrigel with proper medium. Development of organoids has been observed the day after seeding. Organoids have been passaged, expanded and stored in liquid nitrogen to constitute a biobank. For morphological characterization, after reaching an appropriate volume, organoids have been fixed in 4% paraformaldehyde, stained for hematoxylin and eosin (H&E) and immunohistochemistry (IHC) for ki67, CDX2, CK20, MUC2/5 was performed. Immunofluorescence (IF) for DAPI, Lgr5 and CDH1 has been performed.For genomic characterization, DNA has been extracted with phenol-chlorophorm and sequenced with an internal customized panel. A comparison statistical test with matched tissue has been performed.
Results
The success rate in CRC-PDOs establishment is around 80%. PDOs staining with H&E shows a morphology comparable to the original tissue. IHC shows ki67 staining, nuclear positivity for CDX2, cytoplasmic CK20 and MUC2/5. IF evidences the positivity for the intestinal stem cells marker Lgr5 and CDH1 thus confirms that PDOs faithfully recapitulate original tissue morphology and cell composition.Between 80 and 120 ng of genomic DNA has been sequenced. Spectrum of PDOs mutations and polimorphisms displays a good concordance with that of matched patients with an R of linear concordance of 0.9.
Conclusions
We have developed a robust protocol for efficient development of CRC PDOs. Organoids recapitulate morphological and genomic features of the original patient. They work as a solid in vitromodel, suitable for functional studies and drug screening, helping in promoting precision medicine.
Clinical trial identification
Editorial acknowledgement
Legal entity responsible for the study
The authors.
Funding
Grants from the Instituto de Salud Carlos III (PI15/02180 to AC and PI18/01508 to TF). FP is supported by the ESMO 2018 fellowship programme. VG was supported by the ESMO 2014 fellowship programme and by Rio Ortega contract CM18/00241 from the Carlos III Health Institute; TF is supported by Joan Rodes contract 17/00026 from the Carlos III Health Institute. NT was supported by Rio Ortega contract CM15/00246 from the Carlos III Health Institute; DR was supported by Joan Rodes contract 16/00040 from the Carlos III Health Institute. MFGB is supported by a Santiago Grisolia fellowship.
Disclosure
A. Cervantes: Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): Merck Serono; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): Roche; Advisory / Consultancy, Research grant / Funding (institution): Beigene; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): Bayer; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): Servier; Advisory / Consultancy, Research grant / Funding (institution): Lilly; Advisory / Consultancy, Research grant / Funding (institution): Novartis; Advisory / Consultancy, Research grant / Funding (institution): Takeda; Advisory / Consultancy, Research grant / Funding (institution): Astellas; Advisory / Consultancy: Pierre Fabre; Research grant / Funding (institution): Genentech; Research grant / Funding (institution): Fibrogen; Research grant / Funding (institution): Amcure; Research grant / Funding (institution): Sierra Oncology; Research grant / Funding (institution): AstraZeneca; Research grant / Funding (institution): Medimmune; Research grant / Funding (institution): BMS; 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.
Resources from the same session
2674 - Multicenter Phase I Trial of Trastuzumab Emtansine (T-DM1) in Combination with Non-Pegylated Liposomal Doxorubicin (NPLD) in HER2[+] Metastatic Breast Cancer (MBC). THELMA Study
Presenter: Elena López-Miranda
Session: Poster Display session 2
Resources:
Abstract
1398 - Phase 1 study of liposomal formulation of eribulin (E7389-LF) in patients (pts) with advanced solid tumors: primary results of dose-escalation part
Presenter: Noboru Yamamoto
Session: Poster Display session 2
Resources:
Abstract
5818 - Polo-like Kinase 1 inhibitor onvansertib synergizes with paclitaxel in breast cancer carrying p53 mutation
Presenter: Antonio Giordano
Session: Poster Display session 2
Resources:
Abstract
5927 - Phase 1b study of heat shock protein 90 inhibitor, onalespib in combination with paclitaxel in patients with advanced, triple negative breast cancer (NCT02474173).
Presenter: Robert Wesolowski
Session: Poster Display session 2
Resources:
Abstract
1695 - Impact of pertuzumab and T-DM1 on prognosis of HER2-positive metastatic breast cancer (MBC) and factors affecting their efficacy: results from the AGMT_MBC-Registry
Presenter: Simon Peter Gampenrieder
Session: Poster Display session 2
Resources:
Abstract
1742 - Clinical profile and outcome of HER2 positive breast cancer patients with brain metastases treated with HER2 targeted therapy: Real world experience.
Presenter: Prabhat Bhargava
Session: Poster Display session 2
Resources:
Abstract
1846 - Pertuzumab, trastuzumab, and docetaxel for HER2-positive metastatic breast cancer: results of single arm phase IV COMACHI study
Presenter: Norikazu Masuda
Session: Poster Display session 2
Resources:
Abstract
5385 - Anti HER-2 Therapies and Left Ventricular Dysfunction The Renaissance Study
Presenter: ANDRES DANIELE
Session: Poster Display session 2
Resources:
Abstract
3320 - Safety and efficacy of T-DM1 in 128 patients with advanced HER2+ breast cancer: The Royal Marsden experience.
Presenter: Nicolò Battisti
Session: Poster Display session 2
Resources:
Abstract
4540 - Use of trastuzumab emtansine (T-DM1; K) after pertuzumab + trastuzumab (PH) in patients with HER2-positive metastatic breast cancer (mBC): challenges in assessing effectiveness of treatment sequencing in the real world (RW)
Presenter: Thibaut Sanglier
Session: Poster Display session 2
Resources:
Abstract