SCIENTIFIC OBJECTIVES:
Generation and characterization of in vitro 3D organoid models for human primary liver cancer:
Aim 1: model liver cancer by directly culturing human primary liver tumors
Current models for PLC either fail to fully recapitulate tumor histology and architecture or are expensive, time consuming and do not allow for personalised drug testing. During this proposal we established a new 3D in vitro model system for PLC. Based on the current knowledge on organoid cultures, we have managed to establish a system to grow human PLC cells long-term in vitro. Interestingly, the tumor-derived organoids (also termed “tumoroids”) recapitulate the original tumor histology and genetic alterations and are also able to generate tumors in an in vivo xenograft mouse model. Furthermore, we have shown that tumoroids can also be successfully used for drug testing, suggesting their use to devise new targeted therapy as well as personalised treatment strategies.
Aim 2: model human liver cancer progression by genetically engineering human liver cancer in vitro using CRISPR/Cas9 technology
Current models to investigate the role of genes in cancer rely mostly on animal studies, which can be very time consuming and cost demanding, especially if resulting in negative outcomes. To overcome this issue, we set up a protocol for introducing mutations in healthy human liver organoids using the CRISPR-Cas9 technology. Interestingly, after mutating TP53, RNF43 and ZNRF3 either alone or in combination, human organoids undergo genetic alterations and phenotypic changes that partially resemble the ones observed in tumoroids. This data suggests that this system could be used as a screening platform to study gene function/dysregulation during tumorigenesis.
EXPLOITATION AND DISSEMINATION:
These results were published in December 2017 in Nature medicine (Broutier et al., Nature medicine 2017). They will open opportunities to study, for the first time, PLC in a physiologically relevant ex-vivo human system. Moreover, PLC-organoids with a short timescale from establishment to drug screening, could be a relevant in vitro liver cancer system for predicting patient-specific drug responses and creating personalized therapies.
