Generation of glioblastoma patient-derived organoids and mouse brain orthotopic xenografts for drug screening.

Robust patient-derived platforms that recapitulate the cellular and molecular fingerprints of glioblastoma are crucial for developing effective therapies. Here, we describe a chemically defined protocol for 3D culture and propagation of glioblastoma in 3D gliospheres, patient-derived organoids (PDOs), mouse brain orthotopic xenografts (PDOXs), and downstream drug and immunofluorescence assays. This simple-to-follow protocol allows assessing drug sensitivity, on-target activity, and combined drug synergy. Promising therapies can then be validated in PDOXs for translation in precision medicine oncology trials. For complete details on the use and execution of this protocol, please refer to Chadwick et al. (2020) and Patrizii et al. (2018).

Gamboa CM ，Jara K ，Pamarthy S ，Liu L ，Aiken R ，Xiong Z ，Danish S ，Sabaawy HE ... -  《-》

Patient-derived organoids and orthotopic xenografts of primary and recurrent gliomas represent relevant patient avatars for precision oncology.

Golebiewska A ，Hau AC ，Oudin A ，Stieber D ，Yabo YA ，Baus V ，Barthelemy V ，Klein E ，Bougnaud S ，Keunen O ，Wantz M ，Michelucci A ，Neirinckx V ，Muller A ，Kaoma T ，Nazarov PV ，Azuaje F ，De Falco A ，Flies B ，Richart L ，Poovathingal S ，Arns T ，Grzyb K ，Mock A ，Herold-Mende C ，Steino A ，Brown D ，May P ，Miletic H ，Malta TM ，Noushmehr H ，Kwon YJ ，Jahn W ，Klink B ，Tanner G ，Stead LF ，Mittelbronn M ，Skupin A ，Hertel F ，Bjerkvig R ，Niclou SP ... -  《-》

Protocol for derivation of organoids and patient-derived orthotopic xenografts from glioma patient tumors.

Tumor organoids and patient-derived orthotopic xenografts (PDOXs) are some of the most valuable pre-clinical tools in cancer research. In this protocol, we describe efficient derivation of organoids and PDOX models from glioma patient tumors. We provide detailed steps for organoid culture, intracranial implantation, and detection of tumors in the brain. We further present technical adjustments for standardized functional assays and drug testing. For complete details on the use and execution of this protocol, please refer to Golebiewska et al. (2020).

Oudin A ，Baus V ，Barthelemy V ，Fabian C ，Klein E ，Dieterle M ，Wantz M ，Hau AC ，Dording C ，Bernard A ，Michelucci A ，Yabo YA ，Kanli G ，Keunen O ，Bjerkvig R ，Niclou SP ，Golebiewska A ... -  《-》

Glioblastoma Model Using Human Cerebral Organoids.

We have developed a cancer model of gliomas in human cerebral organoids that allows direct observation of tumor initiation as well as continuous microscopic observations. We used CRISPR/Cas9 technology to target an HRasG12V-IRES-tdTomato construct by homologous recombination into the TP53 locus. Results show that transformed cells rapidly become invasive and destroy surrounding organoid structures, overwhelming the entire organoid. Tumor cells in the organoids can be orthotopically xenografted into immunodeficient NOD/SCID IL2RG-/- animals, exhibiting an invasive phenotype. Organoid-generated putative tumor cells show gene expression profiles consistent with mesenchymal subtype human glioblastoma. We further demonstrate that human-organoid-derived tumor cell lines or primary human-patient-derived glioblastoma cell lines can be transplanted into human cerebral organoids to establish invasive tumor-like structures. Our results show potential for the use of organoids as a platform to test human cancer phenotypes that recapitulate key aspects of malignancy.

Ogawa J ，Pao GM ，Shokhirev MN ，Verma IM ... -  《-》

Orthotopic Patient-Derived Glioblastoma Xenografts in Mice.

Xu Z ，Kader M ，Sen R ，Placantonakis DG ... -  《-》