Project description:In this study, pediatric ALL patient-derived xenografts (PDXs) inherently resistant to glucocorticoids were cultured in vitro. The study aims to determine discrepancy in gene expressions between different xeno strains.
Project description:In this study, pediatric ALL patient-derived xenografts (PDXs) inherently resistant to glucocorticoids were cultured in vitro. The study aims to determine discrepancy in gene expressions between different xeno strains. The same xenograft was innoculated into 3 mice. Spleen-harvest xenograft samples were analyzed using microarray.
Project description:Glioblastoma (GBM) patient-derived orthotopic xenografts (PDOXs) were derived from organotypic spheroids obtained from patient tumor samples. To detect whether gene expression profiles of GBM patient tumors are retained in PDOXs, we performed genome-wide transcript analysis by human-specific microarrays . In parallel, we analyzed GBM cell cultures and corresponding intracranial xenografts from stem-like (NCH421k, NCH644) and adherent GBM cell lines (U87, U251). PDOXs show a better transcriptomic resemblance with patient tumors than other preclinical models. The major difference is largely explained by the depletion of human-derived non-malignant cells.
Project description:Most pediatric solid tumor cell lines were established decades ago, so patient-matched organoids and xenografts are not available, limiting their translational utility. Here, we present a new approach to generate triads of patient-matched xenografts, 3-dimensional (3D) organoids, and 2D cultures. We have generated 3D organoids from 135 pediatric tumors, including rhabdomyosarcoma, osteosarcoma, neuroblastoma, Ewing sarcoma, Wilms tumor, and retinoblastoma. We have also generated patient-matched 2D cultures (n=82) and cell lines (n=20). When these 3D organoids, 2D cultures, or cell lines were orthotopically implanted into immunocompromised mice, they replicated the molecular and cellular features of the original patient tumor and xenograft. We have completed 352 single-cell and single-nucleus RNA-sequencing experiments (>2.2 million cells) to confirm that our patient-matched triads preserve tumor cell heterogeneity. All samples are renewable and >3,000 vials have been cryopreserved for unrestricted distribution through the Childhood Solid Tumor Network. We demonstrate the utility of patient-matched triads for identifying therapeutic vulnerabilities, providing a powerful tool for advancing pediatric cancer research.
Project description:Brain tumors are the leading cause of cancer-related death in children. Experimental in vitro models that faithfully capture the hallmarks and tumor heterogeneity of pediatric brain cancer are limited and hard to establish. We present a protocol that enables efficient generation, expansion and biobanking of pediatric brain cancer organoids. Utilizing our protocol, we have established patient-derived organoids (PDOs) from ependymomas, medulloblastomas, low-grade glial tumors and patient-derived xenograft organoids (PDXOs) from medulloblastoma xenografts. PDOs and PDXOs recapitulate histological features, DNA methylation profiles and intratumor heterogeneity of the tumors from which they were derived. We also showed that PDOs can be xenografted. Most interestingly, when subjected to the same routinely applied therapeutic regimens, PDOs respond similarly to the patients. Taken together, our study highlights the potential of PDOs and PDXOs for research and translational applications for personalized medicine.
Project description:Glioblastoma is the most common type of malignant brain tumor among adults. We used single-cell RNA sequencing (scRNA-seq) to analyze the diversity of glioblastoma cells.