NANETS2024 17th Annual Multidisciplinary NET Medical Symposium NANETS 2024 Applied Basic Science (13 abstracts)
Use of patient-derived pre-clinical models to identify new and effective treatments for neuroendocrine tumors
Nilakshi Kulathunga 1 , Zoey Wang 1,2 , Justin Kale 1 , Alberto Lens-Pardo 3 , Rusha Chakraborty 1 , Yan Li 1 , Betty Li 1 , Areeba Qureshi 1 , Emmanuel Cherin 4 , Christine E. Demore 2,4 , Hubert Tsui 1,5 , Rocio Garcia-Carbonero 3 , Julie Hallet 6,7 , Calvin Law 6,7 , Simron Singh 6,8 , David Andrews 1,9 , Hon S. Leong 1,2 & Iacovos P Michael 1,2
1Biological Sciences Platform, Sunnybrook Research Institute, Toronto, ON, Canada; 2Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada; 3Oncology Department, Centro de Oncología Experimental, Grupo de Investigación en Tumores Gastrointestinales y Neuroendocrinos, Instituto de Investigación Sanitaria Hospital 12 de Octubre (imas12), CNIO, UCM, Madrid, Spain; 4Physical Sciences Platform, Sunnybrook Research Institute, Toronto, ON, Canada; 5Laboratory Medicine and Molecular Diagnostics, Sunnybrook Health Sciences Centre, Toronto, ON, Canada; 6Clinical Evaluative Sciences, Sunnybrook Research Institute, Toronto, ON, Canada; 7Department of Surgery, University of Toronto, Toronto, ON, Canada; 8Department of Medicine, University of Toronto, Toronto, ON, Canada; 9Department of Biochemistry, University of Toronto, Toronto, ON, Canada
Background: Neuroendocrine tumors (NETs) arise in different organs and are heterogeneous with limited treatment options. Preclinical models established from patient tumor specimens enable precision oncology by assessing the response to various drug treatments. Herein, we aim to develop patient-derived organoids (PDOs) and xenografts (PDXs) for NETs and use them to identify new treatment approaches.
Methods: We established two pre-clinical models: patient-derived organoids (PDOs) and patient-derived xenografts ex-ovo (PDXovo), using fresh surgical samples from patients with gastroenteropancreatic NETs. To establish PDOs, the fresh tumor tissue sample is disassociated in single-cell suspension, and the cells are seeded in 384-well plates coated with chemically defined hydrogel. To assess the PDOs’ progression and drug response, we use ChromaliveTM non-toxic dyes to monitor the organoids’ metabolic state, ER stress, and apoptosis. High-content imaging of the PDOs is done using an automated spinning disk confocal microscope (Opera Phenix, Revvity), and the image analysis uses customized algorithms. To establish NET patient-derived xenograft ex-ovo (PDXovo) models, we engraft small tumor tissue fragments into the chorioallantoic membrane of avian embryos. High-frequency ultrasound imaging (HF-US) is used to measure changes in tumor volume and vascularity. PDXovos are harvested and characterized at the endpoint using immunostaining and molecular biology approaches.
Results: PDOs and PDXovos were established with a success rate of >95% and a take rate of >90%, respectively. We used NET PDOs from primary tumors (small intestine, pancreas, and cecum) and metastases (liver, lymph node) for high-throughput drug screening of clinically approved drugs alone and in combination with therapies currently used for GEP-NET. We have identified BH3-mimetics that inhibit anti-apoptosis proteins and augment the efficacy of clinically approved therapies for NETs. We established over 160 PDXovos originating from 21 primary tumors and metastases. HF-US scanning of the PDXovos revealed successful tumor growth and vascularization. PDXovos were immunostained for neuroendocrine markers, such as chromogranin A, and markers of proliferation and apoptosis. We found that PDXovos resemble the tumor architecture and morphology of the corresponding patient tumor sample. We are currently evaluating the efficacy of the BH3-mimetics drug combinations using the PDXovo models to corroborate our findings further.
Conclusions: Overall, this study showed that we can reproducibly derive NET preclinical models on a large scale. Moreover, we have identified a new class of drugs for NETs, which we are further evaluating. Utilizing the dual approach of PDOs/PDXovo preclinical models will enable us to identify new potential treatments for patients with NETs.
ABSTRACT ID28678
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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