Patient-derived organoids (PDOs) as a novel in vitro model for neuroblastoma tumours

Abstract

Background Neuroblastoma (NB) is a paediatric tumour of the sympathetic nervous system. Half of all cases are defined high-risk with an overall survival less than 40% at 5 years from diagnosis. The lack of in vitro models able to recapitulate the intrinsic heterogeneity of primary NB tumours has hindered progress in understanding disease pathogenesis and therapy response. Methods Here we describe the establishment of 6 patient-derived organoids (PDOs) from cells of NB tumour biopsies capable of self-organising in a structure resembling the tissue of origin. Results PDOs recapitulate the histological architecture typical of the NB tumour. Moreover, PDOs expressed NB specific markers such as neural cell adhesion molecules, NB84 antigen, synaptophysin (SYP), chromogranin A (CHGA) and neural cell adhesion molecule NCAM (CD56). Analyses of whole genome genotyping array revealed that PDOs maintained patient-specific chromosomal aberrations such as MYCN amplification, deletion of 1p and gain of chromosome 17q. Furthermore, the PDOs showed stemness features and retained cellular heterogeneity reflecting the high heterogeneity of NB tumours. Conclusions We were able to create a novel preclinical model for NB exhibiting self-renewal property and allowing to obtain a reservoir of NB patients’ biological material useful for the study of NB molecular pathogenesis and to test drugs for personalised treatments.

Advances in Engineering Human Tissue Models

“…This system helped them to address questions about the disease in a way that would not have been possible by growing neurons in flat surfaces or by engineering mouse models, due to lack of the necessary niche and interspecies differences, respectively (Lancaster et al, 2013 ; Figure 5A ). Since then, organoids have played a prominent role in enhancing our understanding of various biological phenomena, including development and organogenesis (Karzbrun et al, 2018 ; Trisno et al, 2018 ; Vyas et al, 2018 ; Rossi et al, 2019 ; Shi et al, 2020 ), infectious biology (Forbester et al, 2015 ; Leslie et al, 2015 ; Karve et al, 2017 ; Heo et al, 2018 ; Lamers et al, 2020 ), cancer (Li et al, 2018 ; Nagle et al, 2018 ; Fusco et al, 2019 ; Ooft et al, 2019 ), and other diseases. Among others, the recent study by Sachs et al on long-term expanding lung organoids clearly shows the versatility of these experimental tissue formats in faithfully recapitulating the adult epithelial airway structure and function from both healthy individuals and from patients with cystic fibrosis, lung cancer, and viral infections (Sachs et al, 2019 ).…”

The Promise of Patient-Derived Preclinical Models to Accelerate the Implementation of Personalised Medicine for Children with Neuroblastoma

“…Not only did extensive characterisation of this biobank reveal that the organoids recapitulated patient copy number alterations and mutational signatures but also that patient-specific drug sensitivities were retained. For neuroblastoma, Fusco et al have recently described the development of a panel of six neuroblastoma patient-derived organoids [ 70 ]. Again, the organoids retained the genetic and phenotypic features of the original tumour from which they were derived and were superior to matching spheroid culture, as only the organoids, grown in cell suspension embedded in Matrigel, could reproduce the tumour morphology and architecture.…”

An overview of neuroblastoma cell lineage phenotypes andin vitromodels

“…159 NB heterogeneities, such as phenotype markers and chromosomal aberrations, were also retained in PDTOs. 160 As these technologies of cancer organoids and tumor cells reprogramming evolve, enabling and facilitating their routine use in laboratories is necessary, as they will become essential tools for the generation of patient-derived cell lines that can subsequently be used for the discovery of novel predictive progression markers, and drug-screening toward personalized-medicine strategies.…”

“…159 NB heterogeneities, such as phenotype markers and chromosomal aberrations, were also retained in PDTOs. 160…”