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 ... -  《-》

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 ... -  《-》

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 ... -  《-》

Protocol for generation of and high-throughput drug testing with patient-derived colorectal cancer organoids.

Drug sensitivity testing of patient-derived tumor organoids (PDTOs) is a promising tool for personalizing cancer treatment. Here, we present a protocol for generation of and high-throughput drug testing with PDTOs. We describe detailed steps for PDTO establishment from colorectal cancer tissues, preparation of PDTOs for high-throughput drug testing, and quantification of drug testing results using image analysis. This protocol provides a standardized workflow for PDTO testing of standard-of-care therapies, along with exploring the activity of new agents, for translational research. For complete details on the use and execution of this protocol, please refer to Tan et al.1.

Tan T ，Mouradov D ，Gibbs P ，Sieber OM ... -  《-》

A novel integrated system using patient-derived glioma cerebral organoids and xenografts for disease modeling and drug screening.

A physiologically relevant glioma tumor model is important to the study of disease progression and screening drug candidates. However, current preclinical glioma models lack the brain microenvironment, and the established tumor cell lines do not represent glioma biology and cannot be used to evaluate the therapeutic effect. Here, we reported a real-time integrated system by generating 3D ex vivo cerebral organoids and in vivo xenograft tumors based on glioma patient-derived tissues and cells. Our system faithfully recapitulated the histological features, response to chemotherapy drugs, and clinical progression of their corresponding parental tumors. Additionally, our model successfully identified a case from a grade II astrocytoma patient with typical grade IV GBM features in both organoids and xenograft models, which mimicked the disease progression of this patient. Further genomic and transcriptomic characterization was associated with individual clinical features. We have demonstrated the "GBM-&Normal-like" signature to predict prognosis. In conclusion, we developed an integrated system of parallel models from patient-derived glioma cerebral organoids and xenografts for understanding the glioma biology and prediction of response to chemotherapy drugs, which might lead to a new strategy for personalized treatment for this deadly disease.

Zhang L ，Liu F ，Weygant N ，Zhang J ，Hu P ，Qin Z ，Yang J ，Cheng Q ，Fan F ，Zeng Y ，Tang Y ，Li Y ，Tang A ，He F ，Peng J ，Liao W ，Hu Z ，Li M ，Liu Z ... -  《-》