Clinically relevant orthotopic xenograft models of patient-derived glioblastoma in zebrafish

Abstract

An accurate prediction of the intracranial infiltration tendency and drug response of individual Glioblastoma (GBM) cells is essential for personalized prognosis and treatment for this disease. However, the clinical utility of mouse PDOX models remains limited given current technical constraints including difficulty in generating sufficient sample numbers from small tissue samples and a long latency period for results. To overcome these issues, we establish zebrafish GBM xenografts of diverse origin, which can tolerate intracranial engraftment and maintain their unique histological features. Subsequent single cell RNA-seq (scRNA-seq) analysis confirms significant transcriptional identity resemble invading GBM microtumors observed in the proportionally larger brains of model animals and humans. Endothelial scRNA-seq confirms zebrafish blood-brain-barrier is homologous to the mammalian. Finally, we establish a rapid and efficient zebrafish PDOX (zPDOX) model, which can predict long-term outcomes of GBM patients within 20 days. zPDOX provides a novel avenue for precision medicine of GBM, especially for the evaluation of intracranial infiltration tendency and prediction of individual drug sensitivity.

Zebrafish Cancer Avatars: A Translational Platform for Analyzing Tumor Heterogeneity and Predicting Patient Outcomes

“…The xenografted cells showed significant gene enrichment in categories, such as tumor metastasis, cell migration and epithelial-to-mesenchymal transition. This transcription identity suggests that the zPDX models can recapitulate the supportive tumor microenvironment that patient-derived cells need [95].…”

“…Following the success of xenograft of human cancer cell lines, patient-derived xenografts models in zebrafish have been developed and used to assess different attributes of tumor behavior. zPDX models have since been established in multiple cancer types, including gastric cancer [92], colorectal cancer [73], breast cancer [83], non-small cell lung adenocarcinoma [93], pancreatic ductal adenocarcinoma [94] and others [68,74,95]. Ai et al highlighted the importance of an in vivo system to patient-derived cells by comparing the transcriptomic profile of glioblastoma multiforme (GBM) cells that were xenografted to zebrafish larvae brain to GBM cells grown in vitro in typical cell culture conditions.…”

“…Larval zebrafish PDX could address this issue. While the human cancer cells would still need to survive in a foreign tumor microenvironment, cells are xenografted into larvae directly from the primary tumor after dissociation into a single-cell suspension [68,83,92,94,95,119]. Experiments are completed within 5-10 days.…”

Pediatric low-grade glioma models: advances and ongoing challenges

“…(2010) demonstrated that NF1 knockdown increased ERK signaling and increased OPC proliferation in the developing spinal cord ( 109 ). Orthotopic xenografts and immunodeficient zebrafish models have also been developed for brain tumors ( 110 – 113 ). Although many of the zebrafish xenograft models used glioblastoma cells, Yuan et al.…”

One Host-Multiple Applications: Zebrafish (Danio rerio) as Promising Model for Studying Human Cancers and Pathogenic Diseases

“…Primary GBM (glioblastoma) cells were cultured using three cell culture methods: organoid culture, neurosphere culture, and adherent culture, and then injected into the ZF. A comparison of these three culture types showed that attached culture GBM cells yielded the highest success rate, of 58% (attached cells) versus 25% (neurosphere) and 9% (organoid) [ 21 ].…”